Programme Overview

Hui Si Audrey Koh ^a, Aien He ^a, Junzhe Chen ^a and Weibiao Zhou ^a*

^a Department of Food Science and Technology, National University of Singapore, 117542 Singapore, Singapore
^* Corresponding author. E-mail address: weibiao@nus.edu.sg (W. Zhou)

Presenter email: fsthsak@nus.edu.sg

Background and Objectives
High moisture extrusion has been widely applied for the production of meat analogues. Other than plant proteins, microalgae are a potential raw material. Microalgae biomass has been gaining attention for its health benefits and is a sustainable and scalable source of carotenoids. However, most forms of algae impart an undesirable green colour and fishy odour. A. protothecoides can switch to heterotrophic growth in the presence of high glucose concentration, imparting yellow coloration. This yellowish colour eliminates the limitation of a green extrudate, highlighting the potential of A. protothecoides to be used as a raw material for extrusion.

Materials and Methods
Meat analogues were produced via HME using chickpea proteins and various concentration of microalgae. The effect of microalgae content and moisture content on the textural and structural properties of the extrudates were evaluated using textural profile analysis and scanning electron microscopy. The effect of microalgae addition on the appearance of the extrudates and the specific mechanical energy inputs were also evaluated.

Results & Discussion
Results indicate that the incorporation of microalgae biomass resulted in significant changes to textural properties of the extruded meat analogues. Meat analogues containing microalgae are softer and more tender as compared to meat analogues produced using chickpea protein alone and are more comparable to traditional meat samples. This suggest that the incorporation of microalgae disrupted fiber formation, thus reduced texturing and improved tenderness of the extrudates.

Conclusion
This study highlighted the feasibility of producing extruded meat analogues using chickpea protein with microalgae.

Practical significance
Presently, there exists a technological challenge to achieve meat analogues that meets consumer acceptance in terms of both texture and sensorial attributes. The incorporation of A. protothecoides microalgae biomass appears to be a potential way to improve both nutritional and textural properties of the meat analogues.

Vivian Jing Han Hau^a, Mei Zhi Alcine Chan^a, Yun Kit Ng^a, Shao-Quan Liu^{a, b
a} Department of Food Science & Technology, National University of Singapore, Science Drive 2, Singapore 117542, Singapore
^b National University of Singapore (Suzhou) Research Institute, No. 377 Linquan Street, Suzhou Industrial Park, Suzhou 215123, Jiangsu, China

Email of presenter: vivianhau@u.nus.edu

Background and Objectives
Amid rising demand for alternative proteins, protein-rich Auxenochlorella protothecoides was previously cultivated heterotrophically in nitrogen-supplemented hydrolysates derived solely from food side-streams via a circular bioeconomy approach. To enhance industrial relevance, this study upscaled cultivation to a 6.5-L bioreactor, and characterized cell wall to optimise protein extraction and guide biorefinery strategies in the future, including wholebiomass utilisation.

Materials and Methods
First, cultivation was conducted in a 6.5-L bioreactor using the same cultivation medium as prior flask cultivation, and the growth performance and biomass composition of A. protothecoides were compared. Next, cell wall structure was analysed via electron microscopy while its composition was assessed via monosaccharide and amino acid profiling.

Results & Discussion
Bioreactor cultivation, with precise control of key parameters, yielded a two-fold increase in biomass productivity (11.35 g/L/day) and a three-fold increase in specific growth rate (1.70 day-1) compared to flask cultivation, while maintaining biomass yield (23.19 g/L) and true protein content (28.20% dry mass). Moreover, essential amino acid content was 42.64% total protein, and protein bioaccessibility and bioavailability was 78.81% and 64.81% respectively.

Electron microscopy revealed that A. protothecoides cell wall exhibits recalcitrance and flexibility. Observations further suggest dynamic cell wall development throughout the cell cycle, with a single-layered wall forming as early as the autosporulation stage, which subsequently evolved into a trilaminar structure at later stages. Moreover, preliminary compositional analysis indicated that the cell wall is primarily composed of glucose, mannose and galactose, with further characterisation ongoing.

Conclusion & Practical Significance
Overall, cultivation of protein-rich A. protothecoides in a 6.5-L bioreactor is a crucial step towards industrial production and advanced feeding strategies (e.g., semicontinuous) to further improve productivity. Concurrently, cell wall insights are essential to address protein extraction challenges, guide subsequent biorefinery approaches, or potentially enable whole-biomass utilization via fermentation. Together, these advances would strengthen the industrial viability of A. protothecoides as a sustainable alternative protein.

Mya Myintzu Hlaing, Walter Okelo, Netsanet Shiferaw Terefe CSIRO, Agriculture and Food, Werribee, VIC 3030, Australia
Email: Myintzu.hlanig@csiro.au

As the global demand for protein-rich foods rises, traditional protein sources present significant environmental and resource challenges. This study investigates the potential of Brewer's spent grain (BSG), a by-product of the brewing industry, as a substrate for microbial protein production, offering a sustainable and efficient alternative.

Our research focuses on enzymatic hydrolysis and fermentation of BSG using various microorganisms, including yeast and filamentous fungi. Treatments with customized enzyme cocktails for lignocellulosic materials resulted in a sugar conversion rate of ≥ 90% from total carbohydrates, demonstrating the highest level of conversion among all other physicochemical treatments. This high efficiency promoted robust microbial growth and significant increases in crude protein content.

Preliminary techno-economic analysis indicates that processing BSG into single-cell protein and other protein-rich products is economically viable, with a projected payback period of around four years. This study demonstrates that BSG can be effectively repurposed, contributing to more sustainable and environmentally friendly protein production systems.

These findings highlight the potential for BSG to be a valuable feedstock in the food and feed industries, supporting the development of sustainable bioprocessing technologies. This presentation will detail the enzymatic hydrolysis and fermentation processes, microbial protein yields, and economic feasibility, providing valuable insights for researchers, industry professionals, and policymakers interested in sustainable grain processing and protein production.

Dr. E.L. Ang (SIFBI, A*STAR)
Dr. M.N. Antipina (SIFBI, A*STAR)
Dr. P. Arumugam (SIFBI, A*STAR)
Prof. J.S. Chen (SIFBI, A*STAR)
Ms. X.Y. Chua (SIFBI, A*STAR)
Dr. Y. S/O Kanagasundaram (SIFBI, A*STAR)
Dr. Y.W. Khin (SIFBI, A*STAR)
Dr. N. Khongsay (SIFBI, A*STAR)
Dr. J. Ling (SIFBI, A*STAR)
Dr. T. Manavalan (SIFBI, A*STAR)
Dr. S.B. Ng (SIFBI, A*STAR)
Ms. T.B.V. Nguyen (SIFBI, A*STAR)
Dr. R. Osen (SIFBI, A*STAR)
Dr. X.Y. See (SIFBI, A*STAR)
Dr. M.H. Wong (SIFBI, A*STAR)
Mr. A. Prithiviraj (ARTC, A*STAR)
Dr. F.T. Wong (IMCB, A*STAR)
Mr. Y. Ao (SIMTech, A*STAR)
Ms. S.Y. Choy (SIMTech, A*STAR)
E. Ho (SIMTech, A*STAR)
Prof. W. Zhou (NUS)
Prof. S. Leong (SIT)
Dr. E. Ong (SP)
Dr. M. Weingarten (SIFBI, A*STAR)
Email: Melanie_Weingarten@sifbi.a-star.edu.sg

The Alternative Protein Development Platform (APDP) is a 3.5-year Singapore Food Story grant programme demonstrating the development of novel food ingredients from alternative protein sources (plants, fungi, algae, yeast, etc.) with tailored functionalities for a wide range of food applications (i.e., meat analogs, seafood, and dairy). We are generating a comprehensive alt-protein knowledge platform on protein quality (nutritional, sensory, and functional properties) for ingredients and prototypes. One of the main outcomes is a fully operational pilot-scale development platform (TRL 3-6), capable of translating new research findings into prototypes. The APDP is led by A*STAR’s Singapore Institute of Food and Biotechnology Innovation, and the team includes the NUS, SIT, and SP.

Since 2024, we have built a fully operational lab-topilot Fermentation Joint Lab which is a collaboration between A*STAR’s Singapore Institute of Food and Biotechnology Innovation and Scale-up Bio. Here, we have fermentation and downstream processing capacities supporting up to 200 L scale embedded into Nurasa’s Food Tech Innovation Centre, serving various ecosystem partners and customers from around the world like startup companies, SMEs, MNCs, and Singapore institutes of higher learnings and universities to translate and bring their technology closer to market.

Su Lingshan: NUS, lingshan_su@nus.edu.sg
Jing Linzhi: NUS
Huang Dejian: NUS

Cellular agriculture offers a sustainable alternative to conventional meat production. However, the development of suitable 3D edible scaffolds remains a key challenge. This study aimed to develop food-grade porous scaffolds using cereal-derived prolamins, which are abundant, lowcost, water-stable, and biocompatible. Prolamins, extracted from corn (zein), barley (hordein), rye (secalin), and red sorghum (kafirin), were processed into porous scaffolds by applying commercial sugar cubes as highly permeable templates. The scaffolds exhibited uniform porosity and structural integrity, with preliminary evaluations showing good potential for supporting cell adhesion and growth. Moreover, the cultivated meat prototypes demonstrated promising potential in providing higher protein content and unique mouthfeel and appearance characteristics, highlighting the viability of prolamin in promoting cultivated meat production.

Chua Yu Feng, Quek Jun Ping, Ng Say Kong

chua_yu_feng@bti.a-star.edu.sg

With an ever-growing world population estimated to peak at 10.3 billion in the mid-2080s, there is an increase in the global consumption of aquatic foods at an average rate of 3.0% per year. This increase in demand for food puts a strain in the aquatic wildlife. Cellular agriculture is thus explored as an alternative to conventional seafood production. Cultivated meat offers the potential to mitigate disease risks such as scale drop disease virus (SDDV) and viral nervous necrosis (VNN) that are commonly found in the fish farms. However, one bottleneck in cultivated technologies is that animal cells commonly rely on fetal bovine serum (FBS) for cell growth. As FBS is obtained from calf blood, this requirement necessitates the rearing of cows, which is counter-productive to one objective of cultivated technologies to avoid animal rearing. In addition, FBS poses risks of animal pathogen contamination. In this work, we utilized media component screening to achieve serum-free culture for a commercially available fish cell line. The formulated serum-free media can sustain cell growth for long term culture in both adherent and suspension format. A growth rate of 0.51 cells per day and doubling time of 1.17 days was achieved for the adherent culture whereas growth rate of 0.694 cells per day and doubling time of 0.998 days was observed for the suspension culture.

Shini Yang^1,2, Xiaojun Liao², Zhenzhen Xu^1,2*

E-mail address: xuzhenzhen@caas.cn

Sweet potato (Ipomoea batatas L.) is one of the most widely cultivated food crops worldwide, with annual production surpassing 100 million tons. In addition to starches and soluble sugars, sweet potato roots contain 1.73%–9.14% protein (dry basis), most of which is water-soluble and easily digestible. Nearly half of the global sweet potato yieldis processed into starch, during which large volumes of protein-rich wastewater aregenerated. If not properly treated, this wastewater leads to environmental pollutionandthe loss of valuable bioresources. In this study, proteins were recovered and characterized from yellow powder (YP), a spray-dried byproduct derived from sweet potato starch wastewater. Extraction conditions were optimized using response surfacemethodology (RSM), considering material-to-water ratio, pH, and temperature as independent variables. The optimal extraction conditions were identified as a materialto-water ratio of 50 mg/mL, a pH of 9.5, and a temperature of 30 °C, which together resulted in a maximum protein recovery rate of 61.2%. Proteomic analysis revealedthepresence of 25 proteins, including sporamins, which demonstrated resistance todigestive enzymes and contributed to emulsifying stability. Additional proteins identified, such as starch phosphorylase and polyphenol oxidase, exhibit potential for industrial enzyme applications. Furthermore, superoxide dismutase, an antioxidant enzyme that eliminates reactive oxygen species, was also detected, indicating potential health-related functions. This study demonstrates the potential for transforming agroindustrial side-streams into valuable protein-rich resources. The integrated use of RSMand proteomics provides a practical and sustainable approach for sweet potato starchprocessors to reduce waste, recover functional proteins, and enhance the value of industrial byproducts.

References
[1] Mu, T.H., Sun, H.N., Zhang, M., Wang, C. Sweet Potato Processing Technology, chapter 2: 49-119, 2017.
[2] Yang, S.N., Liu, H.H., Liao, X.J., Kong, X.K., Xu, Z.Z. Journal of Food Science, 87(1): 339-352, 2022

Khai Jien Kong, PhD. (khai@m3triq.com)
Co-founder & CTO

Zi Ying Tan, PhD. (ziying@m3triq.com)
Co-founder & CEO
M3TRIQ

Cultured meat offers a sustainable alternative to traditional livestock farming, but the high cost of essential growth factors such as FGF, IGF, and TGF-β remains a key barrier to commercial scalability. To address this, we introduce M3TRIQ, a modular large language model (LLM) agent designed for autonomous small molecule design, aimed at replacing or mimicking the function of expensive growth factors in cell culture systems.

M3TRIQ integrates multiple tools into a coherent agentic workflow, enabling natural language-driven coordination of cheminformatics, bioactivity databases, docking simulations, and QSAR modeling. Users input desired biological outcomes (for example, “mimic IGF-1 activity in bovine myoblasts”), and the agent autonomously constructs a molecular design pipeline. It retrieves relevant ligands, performs molecular docking, builds predictive QSAR models, and suggests novel compound candidates.

Kong Yan^a, Huang Dejian^a,b

^a National University of Singapore (Suzhou) Research Institute, 377 Lin Quan Street, Suzhou Industrial Park, Suzhou, Jiangsu 215123, PR China
^b Department of Food Science and Technology, 2 Science Drive 2, National University of Singapore, 117542, Singapore

Email address: yan.kong@nusri.cn

Animal gelatin has long been valued in food processingfor its desirable physicochemical and textural properties. However, its thermal instability, batch-to-batch variability, and incompatibility with emerging food manufacturing technologies, such as 3D printing, limit its application in modern plant-based systems. In this study, pumpkin seed protein (PSP) was combined with alginate (ALG) to develop a novel, functional hydrogel system capable of mimicking the texture and rheological behavior of animal gelatin. Through systematic formulation and processing, PSP–ALGhydrogels were engineered to achieve controlled gel strength, elasticity, and visual appearance, thereby meeting the requirements for texture quality modulation in plant-based food products. Mechanical tests showedthat increasing gelatin analog concentration improved hardness (from 0.11 N to 1.46 N) and chewiness (from 0.02N to 0.99 N), with saturation observed beyond 6%. ALGat 2–3% was critical in achieving gelatin-like firmness, while PSP (2–6%) contributed to springiness, cohesiveness, anda gelatin-like color. Rheological and molecular interaction studies confirmed stable PSP–ALG network formation, ensuring performance under printing and post-processing conditions. Notably, the printed Arctic surf clamandvegansea cucumber prototypes displayed realistic color fidelity and biomimetic textures, achieving a convincing seafood like appearance and mouth feel. These findings provide new theoretical insight into the plant based hydrogels and support their application in the production of high-fidelity, sustainable 3D-printed seafood analogs. The work lays a foundation for texture-customizable, plant-based products aimed at enhancing consumer acceptance of next-generation functional foods.

Afsaneh Taheri^a, Chee Fan Tan^b, Xuezhi Bi^b, Fatin Natasha Binte Abdul Halim^a, Juan Du^*a,c,d

^a Food, Chemical and Biotechnology Cluster, Singapore Institute of Technology, 10 Dover Drive, Singapore 138683, Singapore
^b Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (A*STAR), 138668, Singapore
^c Department of Food Science, Purdue University, 745 Agriculture Mall Dr, West Lafayette, IN, 47907, USA
^d Sengkang General Hospital, Singapore Health Services, 110 Sengkang East Way, Singapore 544886, Singapore

Afsaneh.taheritelgari@singaporetech.edu.sg

This study investigated the effects of pH-shifting (acidic/alkaline) and ultrasound treatments on the physicochemical and functional properties of mung bean protein (MBP) and its application in gellan gum (GG)-based complex coacervation gels. The modified MBP was characterized for particle size, ζ-potential, morphology, solubility, secondary structure, and allergenicity. The treated MBP was complexed with GG to evaluate the critical pH values (pHc and pHφ) involved in complex formation. Subsequently, we utilized these complexes to form gel structures and assessed their rheological behaviour and in vitro digestibility through simulated oral, gastric, and intestinal phasesThe combined treatments significantly reduced particle size (174.6 ± 1.52 nm for MBP–12U) and enhanced surface charge (−26.3 ± 0.05 mV for MBP–7U), while structural analysis revealed a shift from β-sheets to α-helix (72 ± 0.25% for MBP–2U). The modified MBP exhibited improved solubility (55-90%), reduced antinutritional factors (e.g., trypsin inhibitory activity), and enhanced flexibility. Reduced Vig r 4 levels in MBP–2U, MBP–12, and MBP–12U were linked to increased free thiol group content, while the shift to an α-helix structure was associated with decreased β-conglycinin levels. The combined treatments affected the electrostatic interactions between MBP and GG, enhancing the pHc (6.25) and pHϕ (6.97) in MBP–7U. Rheological analysis revealed that MBP–2U formed a robust gel network with GG (G′ = 215 Pa), attributed to its high free sulfhydryl content. In vitro digestion of the gel samples demonstrated gastric resistance, with complete hydrolysis in the intestine. These findings highlight the potential of pH-shifting and ultrasound as practical strategies to enhance MBP functionality, enabling the development of plant-based gels with the potential for controlled delivery applications.

Tak Gun Jeremy Chin ^a, Juan Du ^a,b,c

^a Food, Chemical and Biotechnology Cluster, Singapore Institute of Technology, 10 Dover Drive, Singapore 138683, Singapore
^b Department of Food Science, Purdue University, 745 Agriculture Mall Dr, West Lafayette, IN, 47907, USA
^c Sengkang General Hospital, Singapore Health Services, 10 Hospital Boulevard, Singapore, 15 168582, Singapore

Presenter email: jeremy.chin@singaporetech.edu.sg

This study investigated the impact of Hofmeister series anions on amyloid fibrils formation and its subsequent effect on the rheological properties of protein-fibril gels. Protein fibril formation kinetics study using Thioflavin T (ThT) assay showed that the addition of chaotropic anions increased the rate of fibril formation. Fourier transform infrared (FTIR) spectrum results revealed a marked increase in β-sheets in protein secondary structure, which is a sign of amyloid fibril formation. Surface charge measurement of the formed fibrils revealed that chaotropic salts strengthened the electrostatic interactions between protein molecules. Additionally, particle size analysis indicated that the addition of chaotropic salts also enhanced fibril aggregation, leading to the formation of larger particles. Atomic Force Microscopy (AFM) images revealed worm-like structures approximately 400nm in length, with periodic helical features every 15 nm interval indicative of fibril formation. No significant variation in fibril structure was observed between the samples treated with different anions. Rheological properties of gels formed under temperature sweep using 1:1 10% (w/w) mung bean protein-fibril showed that the addition of fibrils significantly increased the viscosity and gel strength of the protein solution, with the strongest gels observed in samples treated with chaotropic salts which is the order of Hofmeister series. Cl- treated samples under frequency sweep tests demonstrated greater gelling stability during the gelation process. This study highlighted the roles of anionic salts in mung bean protein fibril formation and rheological properties of protein-fibril gels, offering valuable insights for applications in food, pharmaceuticals, and biomaterials.

Qifeng Lin, Xin Shan Lim, Renee Chin, Qingsong Lin

Protein and Proteomics Centre, Department of Biological Sciences, National University of Singapore

Email: dbslinq@nus.edu.sg

Background & Objectives
With the global food supply struggling to meet demand, novel food products, such as plant-based meats, are rapidly gaining traction as viable alternative protein sources. Molecular characterization of alternative proteins enables comprehensive nutritional profiling and safety assessment to enhance consumer confidence.

Materials & Methods
We explored different extraction conditions, such as buffer chemical composition and extraction procedure, to develop a protocol for comprehensive protein extraction in alternative protein food matrices. We then performed proteomics profiling to characterize several plant-based meat products.

Results & Discussion
Our optimized protein extraction protocol achieved more than 80% extraction efficiency across various food matrices, including ultra-processed alternative proteins such as plant-based and fungi-based meats. Proteomics analysis of different plant-based meats against their main ingredients revealed distinct proteome profiles, suggesting that food processing methods selectively enrich discrete protein populations. Importantly, key protein classes including bioactive proteins/peptides, flavor peptides and allergens were detected.

Conclusions
Proteomics analysis, driven by comprehensive protein extraction, enables deep characterization of alternative proteins at the molecular level.

Practical Significance
Comprehensive proteomics characterization informs food processing strategies to enhance nutritional, flavor and safety profiles of alternative proteins.

Yujie Lv ^{1, 2}, Weichen Huang², Chaoyue Ge ^{1, 2}, Lianchi Wu ², Zhaoying Hu², Shenao Zhan², Xinyu Shen^{1, 2}, Dongyou Yu^{1, 2, 3}, and Bing Liu^{1, 2, 3}

¹ Hainan Institute, Zhejiang University, Sanya 572000, China
² College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
³ ZJU-Xinchang Joint Innovation Centre (TianMu Laboratory), Gaochuang Hi-Tech Park, Shaoxing 312500, China

Background & Objectives: Polystyrene microplastics (PS) are pervasive environmental pollutants linked to reproductive toxicity in animals. Magnolol (MAG), a natural anti-inflammatory compound, shows therapeutic potential, but its role in mitigating PS-induced oviducal damage remains unexplored. This study aimed to investigate MAG’s protective effects on PS-exposed laying hens, focusing on mucosal barrier restoration, inflammation/apoptosis regulation, and microbiota modulation.

Materials & Methods: A total of 270 Hy-line white laying hens (37-week-old) were divided into three groups: control (CK, basal diet), PS-exposed (PS, basal diet + 1 mg/mL PS gavage for 4 weeks), and MAG-treated (PM, 400 mg/kg MAG-supplemented diet + PS). Egg quality parameters, oviduct histopathology, mucosal barrier markers, microbiota, and inflammatory/apoptotic pathwayswere analyzed. In vitro validation used HD11 macrophages exposed to PS ± MAG.

Results & Discussion: PS exposure significantly decreased albumen and eggshell quality, which are associated with the reduced abundance of the beneficial bacterium Lactobacillus crispatus in the magnum and uterus, impaired mucosal barrier function, and increased mucosal and induced inflammatory responses and apoptosis. MAG effectively ameliorated PS-induced inflammatory damages and apoptosis by inhibiting M1 macrophage polarization. As a result, the oviducal mucosal barrier function was improved evidenced by the increased Claudin and MUC-2 expression. In vitro cell culture experiments also confirmed MAG could inhibit macrophage M1-type polarization via suppressing nuclear factor-κB (NF-κB) signaling pathway activation.

Conclusions: MAG alleviates PS-induced oviduct damage through dual mechanisms: (1) NF-κB inhibition to suppress M1 macrophage polarization and inflammation, and (2) Lactobacillus crispatus-dependent mucosal barrier restoration. This mitigates apoptosis and improves egg quality.

Practical Significance: This study identifies MAG as a cost-effective dietary supplement to counteract microplastic toxicity in poultry production. Its dual action on microbiota and inflammation offers translational potential for improving reproductive health in livestock under environmental pollutant stress.

Ian En Kai Mak¹, Yueying Yao¹, Clarinda Nataria Sutanto¹, Lingyin Yu¹, Zi Ning Leong¹, Chin Meng Khoo², Jung Eun Kim¹

¹Department of Food Science and Technology, National University of Singapore, Singapore
²Department of Medicine, Division of Endocrinology, National University Hospital, Singapore

Presenter’s email: ian.mak@u.nus.edu

Background & Objectives: Higher protein (HP) intakes can deleteriously modulate the gut microbiome, with different HP sources potentially eliciting differential effects. Healthy dietary patterns (HDP) may mitigate these changes, but clinical evidence in older adults remains scarce. This study explored the influence of different sources of HP intakes with a HDP on the gut microbiome in Singaporean older adults.

Materials & Methods: Fifty-five older adults (66 ± 1 y, mean ± SEM) were randomly assigned to either the control (n = 19), casein (n = 18), or soy (n = 18) group. All groups followed a HDP, with the casein and soy groups additionally consuming 20 g/d of casein (animal source) and soy (plant source) protein isolates for 16 weeks. Fecal and serum short-chain fatty acids (SCFAs), plasma and urine uremic toxins, and fecal microbiota composition were assessed at weeks 0, 8, and 16.

Results & Discussion: Total serum SCFA levels significantly increased from week 0 to 16 in the soy group (10.8 ± 5.1 µM, P = 0.047), driven by greater acetic acid levels (11.9 ± 5.2 µM, P = 0.037). Significant time and interaction effects were observed for plasma indoxyl sulfate (IS) (P_Time = 0.023) and p-cresyl sulfate (pCS) (P_Interaction = 0.029), which reduced in the soy group from weeks 8 to 16 (IS: -1.5 ± 0.5 µM, P = 0.022; pCS: -5.6 ± 1.3 µM, P = 0.001). Alpha diversity was reduced from week 0 to 16 overall (P_Time < 0.001), with reductions observed in the soy and control group. No other microbiome changes were noted.

Conclusion: HP intakes with a HDP do not mostly alter the gut microbiome, although protein source may influence microbiome changes.

Practical significance: HP intakes with a HDP can help avoid deleterious microbiome changes linked with HP intakes.

Zi Ning Leong^a, Ian Mak En Kai^a, Chen Yang^a, Du Xiangyi^a, Jung Eun Kim^{a, b}

^a Department of Food Science and Technology, National University of Singapore, Singapore
^b Bezos Centre for Sustainable Protein, National University of Singapore, Singapore

Email of presenter: leong.zining@u.nus.edu

Background: Processing of protein-containing food sources is widely employed to improve their functionality, palatability, and nutritional quality. Numerous studies have reported enhanced protein solubility and reduced anti-nutrient content following processing; however, effects may vary depending on processing method and protein type. Therefore, this systematic review and meta-analysis elucidated the overall effects of processing methods on protein digestibility.

Methods: 10,739 articles were retrieved from PubMed, Embase, CINAHL and COCHRANE. After screening, 245 articles were included in the systematic review, of which 23 human studies were used for meta-analysis. Eligible articles compared processed proteins with less or differentially processed counterparts and reported protein digestibility-related outcomes. Examples of processing methods assessed included extraction, fermentation, heating, germination, extrusion, or hydrolysis, while outcomes assessed included in vitro digestibility (calculated from residual protein content, pH drop) and clinical markers (postprandial amino acids, muscle synthesis rates). Further subgrouping compared effects between animal and plant sources.

Results: Results from in vitro gastrointestinal models revealed varying degrees of improvement in protein digestibility after processing. However, meta-analysis of human studies showed no consistent improvements in digestibility-related outcomes except for a significant increase in peak essential amino acid concentrations (Cmax(EAA)) [WMD: 160.45 uM, 95%CI (7.67, 313.24)]. This was more evident for animal sources [WMD:174.68 uM, 95%CI (12.08, 337.27)] than for plant sources [WMD: 91.03 uM, 95%CI (-475.40, 657.46)].

Conclusions: While processing generally enhances protein digestibility in in vitro models, corresponding improvements observed in human studies were limited, especially for plant proteins. These findings highlight limitations in current processing techniques for optimising plant protein bioavailability, and the challenge of translating in vitro results into physiological benefits. Further refinement of processing is essential to improve protein bioavailability, especially for plant proteins.

Practical Significance: This review provides a comprehensive understanding of processing effects on protein digestibility, offering insights into optimising protein bioavailability through processing.

Xinrui Yin¹, Jiajun Long¹, Tongwei Guan¹, Zheng Zhou¹

¹College of Food and Biological Engineering, Xihua University; Food Microbiology Key Laboratory of Sichuan Province, Chengdu 610039, China

In recent years, plant protein-polysaccharide nanocomplexes have attracted the attention of many researchers as biomaterials with wide application prospects. The purpose of this study was to explore the conditions of different polysaccharides and alcohol-soluble proteins, and to analyze their physicochemical properties. First, the effects of pH, concentration and proportion on the particle size and surface charge of the complex were compared by the combination of barley barley or sorghum melin and polysaccharide (carrageenan and pectin). The nanocomposites with good particle size, dispersion and encapsulation properties were prepared. Then the stability of the alcohol-soluble protein-polysaccharide nanocomplex was compared under different temperature and salt ion strength. Subsequently, the structure of the nanocomposites was analyzed by Fourier transform infrared spectroscopy (FT-IR), circular dichroism (CD) and endogenous fluorescence analysis, and the encapsulation efficiency of the bioactive substances was studied. In this study, alcohol-soluble protein-polysaccharide nanocomplexes were constructed successfully and their physicochemical properties were analyzed comprehensively. The nanocomplex has good thermal stability, salt ion temperature and encapsulation ability, which provides a theoretical basis for the application of alcohol-protein-polysaccharide nanocomplex in food, medicine and other fields, and lays a foundation for subsequent research.

Acknowledgments
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Thimo Ruethers^1-4, Shaymaviswanathan Karnaneedi^3,4, Afifah Sean^1,3, Michael G. Leeming⁵, Renee Chin^1,3, Roni Nugraha^1,6, Vachiranee Limviphuvadh², Nicholas A. Williamson⁶, Dean R. Jerry¹, Dianne E. Campbell^4,7,8, Sebastian Maurer-Stroh², Anand K. Andiappan⁹, Andreas L. Lopata^1-4

¹ Tropical Futures Institute (TFI), James Cook University (JCU), Singapore
² Bioinformatics Institute (BII), Agency for Science, Technology and Research (A*STAR), Singapore
³ Molecular Allergy Research Laboratory (MARL), JCU, Australia
⁴ Centre for Food and Allergy Research (CFAR), Australia
⁵ Bio21 Institute, University of Melbourne, Australia
⁶ Department of Aquatic Product Technology, IPB University, Indonesia
⁷ Department of Clinical Immunology and Allergy, Children's Hospital at Westmead, Australia
⁸ Faculty of Medicine and Health, University of Sydney, Australia
⁹ Singapore Immunology Network (SIgN), A*STAR, Singapore

Background & Objectives
Innovations in cellular agriculture are reshaping the global food landscape, offering sustainable alternatives to conventional animal protein. Cultivated seafood is emerging as a promising solution to environmental degradation, overfishing, and food security challenges. However, it may pose underexplored human health hazards. Seafood allergies are a mounting global public health concern, requiring careful considerations in safety assessments of muscle cell-based foods as many muscle proteins are potent allergens. Seafood, including fish, is a leading trigger of food-induced (fatal) anaphylaxis globally. Accordingly, food safety concerns remain a critical barrier to market approval and integration. Our research investigates how food safety investigations, particularly those addressing allergenicity, promote consumer health and drive adoption of cultivated seafood products.

Materials & Methods
Using a multi-omics approach, we assessed allergy risks in cell-cultivated Japanese eel (Anguilla japonica), a highly sought-after but endangered fish species.

Results & Discussion
Cultivated eel demonstrated >10-fold reduction in all 12 WHO/IUIS-recognised fish allergens and >1,000-fold decrease in the major allergen parvalbumin, with zero IgEbinding detected in 20 fish-allergic individuals—indicating a dramatically reduced allergy risk. Replacing fetal bovine serum with serum-free media eliminated the introduction of non-fish allergens, improving food safety further.

Conclusions
In addition to compliance with requirements for food safety assessments, this targeted approach may offer pathways for liberating diets of allergic individuals who are generally advised to strictly avoid consuming all seafood and its products for a lifetime. Beyond allergen reduction, cultivated seafood appears generally safer and healthier compared to conventional seafood due to the guaranteed absence of environmental toxins, heavy metals, parasites, and antibiotics that frequently contaminate wild and farmed seafood.

Practical Significance
Our study provides the first allergenomic foundation for safety assessments, suggesting that allergy risks are diminished in cell-cultivated fish. As we seek to accelerate the transition to more sustainable food systems, thorough food safety and allergenicity evaluations are central.

Xie Xiaofang, National University of Singapore, xiaofang@nus.edu.sg

Li Zhaofeng, Jiangnan University

Huang Dejian, National University of Singapore

Background & Objectives
D-Allulose is a rare, low-calorie sugar with 70% the sweetness and only 0.3% the energy of sucrose. It is mainly produced by isomerizing D-fructose using D-allulose 3- epimerase (DAE). However, low expression levels and poor enzyme performance have limited its industrial use. This study aimed to achieve high-level expression of a DAE in a food-grade system and evaluate its enzymatic properties for industrial application.

Materials & Methods
A dae gene from a clostridial bacterium (CDAE, GenBank: SABL01000019.1) was cloned to constructed recombinant strain Bacillus subtilis WB600 (pP43NMK-dae). The enzyme was purified using nickel-affinity chromatography (ÄKTA Pure system, GE, USA), and its activity and catalysis products were analyzed via HPAEC-PAD. Statistical significance (P < 0.05) was determined using one-way ANOVA.

Results & Discussion
CDAE was successfully expressed in a food-grade Bacillus subtilis system, reaching intracellular and extracellular activity levels of 222.55 U/mL and 155.50 U/mL, respectively—both exceeding those reported for most other DAEs. Biochemical characterization of the purified enzyme revealed excellent thermostability, an optimal activity at weakly acidic pH, and superior catalytic efficiency. Structural analysis showed a characteristic (α/β)8-barrel subunit and strong interfacial interactions. Additionally, CDAE facilitated D-allulose synthesis from D-fructose and D-glucose via borate-assisted isomerization, achieving enhanced yields of 63.5% and 45.0%, respectively.

Conclusions
CDAE was efficiently expressed in a food-grade Bacillus subtilis system and exhibited excellent thermostability, favorable pH tolerance, and high catalytic efficiency. Its robust enzymatic properties and high-yield D-allulose production from D-fructose and D-glucose demonstrate its strong potential as a biocatalyst for industrial-scale Dallulose manufacturing.

Practical Significance
This study establishes CDAE as a highly promising biocatalyst for industrial D-allulose production, overcoming previous challenges related to low expression and suboptimal enzyme characteristics. Its enhanced expression and catalytic efficiency support its application in the cost-effective, large-scale manufacturing of this low-calorie sugar alternative.

Nicole Min Yee Wong^1#*, Marcus Ting^1#, Jung Eun Kim^{1, 2}

¹ Department of Food Science & Technology, National University of Singapore, Singapore
² Bezos Center for Sustainable Protein, National University of Singapore, Singapore

^#These authors contributed equally to this work

^*Presenter email: nicolewong@u.nus.edu

Background & Objectives:
With the global population expected to reach 9.7 billion by 2050, the demand for protein is anticipated to surge by up to 78%. However, conventional animal and plant protein sources pose sustainability concerns due to high resource use and environmental impact. Microalgae, a promising alternative, are gaining attention for their favourable protein and amino acid content, and their robust growth in varied environments. This review thus aimed to comprehensively evaluate the quantity, quality, and sustainability of microalgae protein compared to conventional sources.

Materials & Methods:
This review consolidated current literature on the protein types, content, and quality of key microalgae species, with comparisons against conventional protein sources. It also examined the effects of cultivation conditions, growth phases, and species on variations in protein content, including comparisons across different cultivation, harvesting, and processing methods.

Results & Discussion:
Microalgae contain up to 70% of protein (dry matter), outperforming that of beef (40%) and soybeans (36%). Their amino acid profiles are generally well-balanced, with amino acid scores (AAS) typically ≥ 1.0, although their protein-digestibility corrected AAS (PDCAAS) may vary. For instance, Arthrospira sp. (PDCAAS 0.8) demonstrates higher digestibility than Acutodesmus sp. (PDCAAS 0.3), likely attributed to the greater cell wall rigidity of the latter, which hence requires processing to improve bioaccessibility. Other factors influencing protein yield and quality include cultivation techniques, harvest phases, and processing methods. Ecologically, microalgae require much less land (0.5 to 1 m2/kg protein) and freshwater (240-2300 L/kg protein), performing comparably to or better than conventional protein sources.

Conclusions:
Microalgae offer a promising and sustainable alternative protein source, characterized by high protein content and quality. However, further research is needed to optimize species selection, improve human digestibility, and enable scalability for large-scale production.

Practical Significance:
Understanding and optimizing microalgae protein production may support the growing global dietary demand sustainably, particularly in the context of increasing environmental and nutritional challenges.

Wei Han Foo^a, Lin Chen ^a*

^a School of Chemistry, Chemical Engineering and Biotechnology, Nanyang Technological University, 62 Nanyang Dr, 637459, Singapore.

Email: fooweihan333@gmail.com

Sustainable and efficient protein alternatives are critical to meet global nutritional demands amid escalating environmental challenges. Chlorella pyrenoidosa, Arthrospira (Spirulina) platensis, and Chlorella sorokiniana—are increasingly recognized for their high protein content, rapid biomass productivity, and low resource requirements. These microalgae are classified as Generally Recognized As Safe (GRAS), making them suitable for food and nutraceutical applications. This study investigates ultrasound-assisted alkaline extraction as a green, scalable strategy for recovering functional proteins from these species for food and nutraceutical use.

An initial comparison of ultrasound mechanisms was performed using a mild alkaline medium (0.01 M NaOH) to preserve native protein structure while enabling solubilization. Ultrasonic probe treatment (30–35 kHz, 5 min) consistently outperformed ultrasonic bath (15 min) across all species. A. platensis achieved the highest recovery (41.11 ± 0.69%) and yield (155.83 ± 2.56 mg/g), while C. sorokiniana showed the greatest relative gain, with yield increasing from 48.62 ± 1.39 mg/g to 66.80 ± 1.51 mg/g. Microscopic imaging confirmed enhanced cellular disruption following probe sonication, validating its superior cavitation efficiency, especially in species with rigid cell walls.

Building on these findings, a response surface methodology (RSM) optimization phase will assess the interactive effects of amplitude, duty cycle, and sonication duration on extraction efficiency. Based on existing literature, A. platensis is expected to exhibit high water and oil absorption capacities (up to 5.0 g/g and 10.0 g/g, respectively) and foam stability (>85%) due to its ordered protein structure. C. pyrenoidosa may achieve strong emulsifying activity (up to 3000 mL oil/g) and foam capacity (~300%), while C. sorokiniana is predicted to demonstrate notable improvements in solubility and emulsion stability.

Collectively, this work supports the development of cleanlabel, high-functionality protein ingredients and advances the role of microalgae as a sustainable protein platform for future food systems.

Wang Haijie^1,2, Zhou Weibiao^1,2
haijiewang22@u.nus.edu

¹Department of Food Science and Technology, National University of Singapore, Singapore
²Bezos Centre for Sustainable Protein at the National University of Singapore, Singapore

Background & Objectives
Scaling up cultivated meat production requires optimization of hydrodynamic conditions to maintain cell viability and productivity. This study employs CFD to analyze how flow behavior and shear stress evolve during bioreactor scale-up and to guide design optimization.

Materials & Methods
The fluid dynamics of stirred-tank bioreactors of 100 mL, 500 mL, and 2 L were modeled. The standard k-epsilon turbulence model was applied. Key parameters—including velocity fields, shear stress, energy dissipation, and Kolmogorov length scales—were evaluated across scales.

Results & Discussion
Results revealed that scale-up leads to intensified shear near impeller tips and increased stagnant zones, particularly at vessel bottoms and corners. Kolmogorov scales decreased with volume, indicating more aggressive microscale turbulence. These shifts suggest potential risks to sensitive myoblast cultures under direct geometric scaling.

Conclusions
CFD revealed non-linear changes in hydrodynamics during scale-up, underscoring the need for design modifications to preserve favorable cell culture environments.

Practical Significance
This work provides a CFD-driven framework for optimizing scale-up in cultivated meat production, offering actionable insights for bioreactor design and operating strategies.

Jiaqi Mi^a,b, Zhini Guo^a, Hanqi Qian^a, William R. Birch^c, Vinayaka Srinivas^d, Weibiao Zhou^a,b*

^a Bezos Centre for Sustainable Protein at the National University of Singapore, 2 Science Drive 2, 117542, Singapore
^b Department of Food Science and Technology, National University of Singapore, 2 Science Drive 2, 117542, Singapore
^c 21 Jalan Maya 3/8, 79100, Johor, Malaysia
^d Happy Petto Pte Ltd, Unit 1-68, 25 International Business Park, 609916, Singapore

Background & Objectives
Cultivated meat (CM) offers a sustainable protein alternative compared to animal meat, but widespread commercialisation is hindered by the high cost of cell culture media manufacturing and ethical concerns of using fetal bovine serum (FBS). In this study, we investigated bioactive plant protein hydrolysates (PPHs) as food-grade, cost-effective alternatives to FBS in cell culture media (CCM), aiming to support muscle cell growth, differentiation, and CM formation on edible scaffolds.

Materials & Methods
PPHs are enzymatically derived from legumes and cereals using food-grade proteases with different substrate preferences under varied hydrolysis conditions. PPHs were characterised by degree of hydrolysis and molecular weight. PPHs were supplemented into the basal media, with or without FBS. Porcine satellite cells were cultured in both 2D format and 3D edible plant-based scaffolds. Amino acid (AA) metabolism was evaluated, cell proliferation in scaffold was evaluated using CCK8 assay. Cryosection samples were used for confocal imaging, and texture profile analysis was conducted to assess the mechanical properties of the CM.

Results & Discussion
Benchmarking against standard growth media with 10% FBS, a range of PPHs enhanced the ability of basal media to maintain PSCs viability without serum. Selected PPHs supported the proliferation, maintained differentiation capabilities of PSCs under low-serum conditions, and promoted cell attachment and proliferation in 3D culture. PPHs significantly altered cellular AA metabolism, enabling enhanced cell growth. A cultivated pork prototype with 16% protein content was developed, and serum usage was reduced by up to 70%.

Conclusions
PPHs represent a scalable, sustainable solution to reduce serum dependence in CM manufacturing, maintaining muscle cell performance while significantly lowering production costs.

Practical Significance
The scalable and cost-effective production of PPHs, as a sustainable food-grade serum-reducing biomaterial, highlights their industrial potential for transforming CM manufacturing and advancing sustainable biomanufacturing.

Yuheng Wang, Hichang Cho, Junhan Fu, Weiyu Zhang

As a country highly dependent on food imports, Singapore has prioritized food innovation to enhance national food security, becoming the first in the world to approve cultivated meat for commercial sale. Despite strong government support, public adoption remains uncertain. Existing research largely focuses on Western populations, leaving a gap in understanding consumer responses in multicultural Singapore. This study examines whether microtargeted messages—tailored to individuals’ top concerns or psychological traits—can more effectively promote cultivated meat than generic or mismatched messages.

We conducted an online survey experiment with 1,000 Singaporean adults recruited via national quota sampling. Participants were randomly assigned to receive either a broad message, a microtargeted message matched to their primary concern, or a mismatched message. Messages were framed around five themes: health, environment, animal welfare, familiarity, and technology safety. Key outcomes—including perceived risks, benefits, knowledge, and behavioral intentions—were measured before and after message exposure. Participants’ traits, such as openness, health consciousness, etc., were assessed using validated scales.

Results showed that microtargeted messages produced the highest average improvements across all outcomes. Compared to broad messages, they significantly increased behavioral intentions (p = .003) and marginally improved attitudes (p = .050). A planned contrast revealed a graded pattern of effectiveness: microtargeted > non-matched > broad. Psychographic targeting was effective only for individuals with high trust in science or strong environmental and health consciousness, who showed significant gains in knowledge and meat reduction. Other traits, such as animal welfare concern or openness to new foods, showed no effect.

These findings suggest that issue-based microtargeting enhances persuasion, and personal traits like science trust and environmental concern can serve as effective targeting dimensions. These findings offer practical guidance for designing effective communication strategies to promote cultivated meat specifically, as well as other novel food technologies, in diverse and multicultural societies.

Siyong You^abc, Yuhan Wan^a, Linzhi Jing^ac, Dejian Huang^abc

^a Department of Food Science and Technology, National University of Singapore, 2 Science Drive 2, Singapore, 117542
^b Bezos Centre for Sustainable Protein, Faculty of Science, National University of Singapore, 2 Science Drive 2, Singapore 117542
^c National University of Singapore (Suzhou) Research Institute, 377 Linquan Street, Suzhou, Jiangsu, China, 215123

Email: e0983555@u.nus.edu

The global aging trend has heightened societal focus on elderly health, particularly dysphagia as a critical concern. To address this, we developed 3D-printable, nutrient-rich food-grade emulsion inks designed for dysphagia diets. Two preparation methods—direct mixing (DM) and sequential addition (SA)—were employed to formulate secalin-xanthan gum gel inks with varying oil contents. The inks were characterized for rheological properties, microstructure, FTIR, LF-NMR, bioaccessibility, and printing performance, with swallowing safety assessed via IDDSI testing. Results demonstrated that SA-derived inks exhibited superior rheological behavior, enhanced lutein bioaccessibility, and more uniform oil droplet distribution compared to DM inks. Notably, the SA5 formulation achieved optimal printability and met the IDDSI Level 4 standard (safe for swallowing). This improvement was attributed to the SA method’s ability to create a denser network structure, effectively encapsulating oil droplets and nutrients. Our findings highlight that tailored oil content combined with the SA method significantly enhances ink performance. This study advances the application of plant-based protein as 3D printing inks in dysphagia-friendly food design and offers a framework for future functional food development.

Ding Xiang Chew

Singapore Institute of Food and Biotechnology Innovation (SIFBI), Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos, Singapore 138669, Singapore.

chew_ding_xiang@sifbi.a-star.edu.sg

Background & Objectives
Hydrocolloids are frequently employed to modify the texture of meat analogues. While the individual functionalities of soy protein isolate (SPI) and konjac glucomannan (KGM) are well-documented, their synergistic interactions remain underexplored. Such interactions are crucial as they dictate the final product's microstructure and texture.

Materials & Methods
SPI-only and SPI+KGM samples were processed in a closedcavity rheometer (CCR), where temperature and shear were controlled. The regime of the process was as follows: measure viscosity at 60 °C, heat to 110 or 130 °C at 1 or 50 % strain, allow samples to cool to 60 °C with no shearing, measure viscosity, and conduct amplitude sweep. Microstructural analysis was performed on processed samples obtained from the CCR.

Results & Discussion
Viscosity change: after processing, the viscosity of all samples increased. SPI-only increased slightly more than SPI+KGM.

Changes during heating: when heated from 60 °C to the target temperatures (110 or 130 °C), the decrease in viscosity of SPI and SPI+KGM samples was similar, indicating that KGM may not exert influence during heating. Yet the result after cooling was different as seen in the amplitude sweep.

Amplitude sweep: SPI+KGM samples had a lower critical strain and lower cross-over point, indicating a microstructure that is more easily broken down. However, SPI+KGM samples had higher storage moduli than SPI-only samples. The combination of lower strain and higher storage modulus could mean that the SPI+KGM samples were more brittle.

Microstructure observations revealed that the addition of KGM caused more fibre-like microstructure than SPI-only.

Conclusions
KGM impacts the rheological and structural properties of SPI systems. While initial heating responses were similar, KGM-containing samples exhibited a more brittle and a more pronounced fibre-like microstructure compared to SPI alone.

Practical Significance
The study highlights KGM's role as a texturising agent in plant-based protein systems.

Hongbin Lin^a,b,*,1, Jianhua Zhao^a,1, Yuqing Xie^a, Jie Tang^a,d, Qin Wang^c, Jie Zhao^a,b, Min Xu^a,b, Ping Liu^a,b

^aSchool of Food and Bio-engineering, Xihua University, Chengdu 610039, China
^bChongqing Key Laboratory of Specialty Food Co-Built by Sichuan and Chongqing, Chengdu 610039, China
^cDepartment of Nutrition and Food Science, University of Maryland, College Park, MD 20742, USAdFood Microbiology Key Laboratory of Sichuan Province, Chengdu 610039, China
¹Hongbin Lin and Jianhua Zhao contributed equally to this work and should be considered co-first authors.

Oxidative stress will occur in cellular metabolism when the accumulated intracellular reactiveoxygen species (ROS), including hydroxyl radicals (OH^-), superoxide anion radicals (O₂^-), singlet oxygen (¹O2), and hydrogen peroxide (H₂O₂), surpass the tolerance of thecellular antioxidant defense system^[1]. This can result in oxidative damage tocellular macromolecules, including carbohydrates, lipids, protein, and DNA^[2]. Oxidativedamage is a main risk factor for a battery of chronic non-communicable diseases ^[3]. Currently, bioactive peptides have received significant attention due to their abundant activities. Among which antioxidant peptides could improve human health by preventingchronic non-communicable diseases such as cardiovascular diseases, cancer, or diabetes ^[4], and are highly promising functional factors ^[5]. This study aimed to investigate theantioxidative and cytoprotective activity of antioxidant peptides from fermented broad beanpaste (FBBP) and explore their potential molecular mechanisms using a combined in silicoand in vitro approach. Seven novel antioxidant peptides (VSRRFIYYL, SPAIPLP, PVPPPGG, KKDGYWWAKFK, LAWY, LGFMQF, and LPGCP) identified by integrated approaches of peptidomics and in silico bioinformatic analysis were synthesized, exhibiting strongantioxidant potential against in vitro radicals. Molecular docking results suggested that thesepeptides could form stable hydrogen bonds and solvent-accessible surface with key aminoacid residues of Keap1, thus potentially regulating the Keap1-Nrf2 pathway by occupying theNrf2-binding site on Keap1. Additionally, they exhibited strong cellular antioxidant activityand could protect HepG2 cells from AAPH-induced oxidative injury by reducing reactiveoxygen species and MDA accumulation. This study firstly unraveled the molecular mechanisms of antioxidant peptides from FBBP, and provided a new theoretical basis for thehigh-value utilization of FBBP.

Acknowledgments
The authors sincerely thank all the panelists for experimental support. This work was supported by the Science and Technology Department of Sichuan Province, China(2020YFN0151, 23ZDYF3100) and Chongqing Science and Technology Commission (cstc2021jscxcylhX0014).

1. Lingjiao Z ,Hanguo X ,Xi H , et al. Food Research International, 2022, 157111266-111266. 2. Likun R ,Yang Y ,Chunmin M , et al. Food Research International, 2022, 162(PB): 112108-112108. 3. Zhao W ,Luo Q ,Pan X , et al. Journal of Functional Foods, 2018, 47503-511. 4. Montoya-Rodríguez A ,Mejía D G E . Food Research International, 2015, 77204-214. 5. Zhen X ,Jianyin M ,Bingbing C , et al. Food Research International, 2022, 157111359-111359.

Qingyan Guo^a,b, Jingjing Zhao^a, Jiabao Peng^a, Yukun Huang^a,b, Bing Shao^a,c

^aSchool of Food and Bioengineering, Food Microbiology Key Laboratory of Sichuan Province, Xihua University, Chengdu, 610039, China
^bChongqing Key Laboratory of Speciality Food Co-Built By Sichuan and Chongqing, Chengdu 610039, China ^cBeijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Prevention and Control, Beijing, 100013, China

Huangshui (HS) is the main by-product during the fermentation process of Baijiu production, which possesses abundant organic matter and microbial resources, rendering it valuable for high-value utilization[1,2,3]. This study collected HS samples from three production workshops with distinct environments, aiming to assess the impact of the brewing microenvironment on the quality of HS and to screen bioactive peptides from HS. The results revealed elevated lactic acid content in HS samples, ranging from 7878.09 to 10531.56 mg/L. Esters, alcohols, and acids were the main flavor compounds, with 32 key volatile compounds identified. Lactobacillus emerged as the predominant bacterial genus in HS, while Cladosporium and Aspergillus dominated the fungi. This indicates that HS has great potential for developing bioactive components. Redundancy analysis (RDA) indicated that the physicochemical parameters of HS were pivotal factors influencing the microbial community. Spearman correlation analysis exhibited significant positive correlations of Cladosporium and Aspergillus with 13 and 3 flavor compounds, respectively, suggesting they may represent the core functional microbial genus responsible for the characteristic flavor of HS. Moreover, HS demonstrated antioxidant and antibacterial activities[4,5], further augmenting post-enzymatic hydrolysis. This result corresponds to the increase in DH of HS after protein hydrolysis and the decrease in protein concentration. The results indicate that the enzymatic hydrolysis process further converts the proteins in HS into compounds such as peptides that have antioxidant and antibacterial activities, providing insights for the comprehensive development and utilization of HS. These findings lay the groundwork for screening functional strains and bioactive peptides derived from HS, laying a theoretical foundation for the application of HS in the food industry[6,7]. However, efficient separation strategies for functional microorganisms and bioactive compounds necessitate further exploration[8,9].

Acknowledgments
This work was supported by the Natural Science Foundation Program of Sichuan Province [Grant No.2023NSFSC1205] and the Talent Introduction Project of Xihua University [Grant No. Z211045].

1. J. Huo, J. Wu, M. Zhao, et al. International Journal of Biological Macromolecules, 161 (2020), pp. 406-416.
2. J. Huo, J. Wu, M. Zhao, et al. Food Chemistry, 330 (2020), Article 127257.
3. Q. Zhao, Z. Lv, M. Zhang, et al. Liquor-making Science & Technology (4) (2022), pp. 78-81.
4. J. Huo, Q. Liao, J. Wu, et al. International Journal of Biological Macromolecules, 223 (11) (2022), pp. 595-605.
5. H. Xu, Z. Wu, D. Zhao, et al. International Journal of Biological Macromolecules, 214 (4) (2022), pp. 212-219.
6. A. Li, G. Liu, L. Li, et al. Food and Fermentation Industries, 45 (2) (2019), pp. 118-123.
7. R. Yang, J. Zhou. Liquor-making Science & Technology (3) (2008), pp. 90-92.
8. Y. Tan, H. Du, H. Zhang, et al. Microbiology Spectrum, 10 (5) (2022), Article e01844.
9. Y. Xu, X. Qiao, L. He, et al. Frontiers in Microbiology, 14 (2023), Article 1324722.

Shuqi Li¹, Clarinda Nataria Sutanto¹, and Jung Eun Kim^1•2

¹Department of Food Science and Technology, Faculty of Science, National University of Singapore, Singapore
²Bezos Center for Sustainable Protein, National University of Singapore, Singapore

E1124672@u.nus.edu

Background: Poor sleep quality is prevalent in older adults, affecting overall quality of life. Dietary factors, particularly dietary protein and certain micronutrients, have been implicated in the regulation of sleep however, their intake in individuals with different sleep quality conditions have not been examined. Therefore, this study aimed to compare dietary factors in Singapore middle-aged and older adults with different sleep quality conditions.

Methods: 104 participants (aged 50-75 years) were recruited, and their sleep quality was assessed by Pittsburgh Sleep Quality Index. Global sleep score (GSS) was further calculated to identify good (GSS S 5) and poor (GSS > 5) sleepers. Dietary intake was assessed by 3-day food record and nutrients that associated with sleep including protein, magnesium (Mg), vitamin B6, B9 and B12 were analyzed.

Results: 38 participants were defined as poor sleepers (37 %), experiencing a longer sleep latency (good vs poor: 10.0 ± 7.5 min vs 26.5 ± 17.7 min, P < 0.001) and a lower sleep efficiency (good vs poor: 93.8 ± 5.8 % vs 84.6 ± 9.9 %, P < 0.001) than good sleepers. No differences were observed in total (good vs poor: 83.6 ± 40.1 g/day vs 78.9 ± 24.4 g/day), animal (good vs poor: 43.9 ± 20.2 g/day vs 45.3 ± 17.2 g/day) and plant protein intake (good vs poor: 37.6 ± 34.6 g/day vs 31.8 ± 14.1 g/day) between good and poor sleepers. There were also no differences in micronutrients intakes. Additionally, no correlations between nutrients intake and sleep quality parameters were observed.

Conclusion and Practical Significance: The findings may suggest lack of associations between dietary factors and sleep quality in Singapore middle-aged and older adults. However, caution is warranted in interpretation due to relatively small sample size, self-reported dietary and sleep data and other potential confounding factors.

Xingyu ZHU^a, Yee Kiat SOH^a, Han Ming YONG^a, Jeremy Kah Sheng PANG^b, Boon Seng SOH^b, Eric Chun Yong CHAN^a

^a Department of Pharmacy and Pharmaceutical Sciences, Faculty of Science, National University of Singapore, 18 Science Drive 4, Singapore 117543, Singapore
^b Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, 61 Biopolis Drive, Proteos, Singapore, 138673, Singapore

Email contact: yonghanming@u.nus.edu

Glycogen Synthase Kinase 3 (GSK-3) inhibitors are increasingly employed in cultured meat production to augment the proliferative capacity of animal-derived muscle stem cells. However, GSK-3 inhibition has been implicated in the modulation of cardiac rhythms in vivo, raising concerns about potential proarrhythmic effects if such compounds persist as residues in cultured meat products. This highlights a critical need for a rigorous cardiotoxicity assessment of GSK-3 inhibitors to ensure consumer safety. Here, we developed a human embryonic stem cell-derived cardiomyocyte (hESC-CM) platform amenable to detecting cardio-electrophysiological perturbations. Using a modified GiWi hESC differentiation protocol, we yielded virtually pure hESC-CMs (93.3% ± 6.8%) expressing cardiac markers SIRPα and TNNT2. These hESC-CMs were also electrophysiologically active, displaying spontaneous beating activity with a mean beat-to-beat (BTB) interval of 2.20 s and a baseline variance of 0.11. Following treatment with danazol, a known cardiotoxicant, the BTB variability significantly increased (BTB variance = 0.20, p < 0.0001), validating the model’s sensitivity to xenobiotic-induced electrophysiological perturbations. Subsequently, we evaluated the effects of CHIR-99021, a widely used GSK-3 inhibitor, on BTB dynamics to assess its proarrhythmic potential. In conclusion, our findings provide important insights into the cardiac safety profile of GSK-3 inhibitors and establish hESC-CMs as a viable preclinical platform for evaluating the electrophysiological risks of xenobiotic residues in novel food products.

Liang Yuxin, MSc¹; Yao Yuanhang, PhD²; Kim Jung Eun, PhD, RD^1,3*

¹Department of Food Science and Technology, National University of Singapore, Singapore
²School of Public Health, Southeast University, Nanjing, Jiangsu, China
³Bezos Center for Sustainable Protein, National University of Singapore, Singapore
^*Corresponding author

Email: yuxinliang@u.nus.edu

Background & Objectives: Carotenoids, naturally found in fruits and vegetables, have antioxidant property. However, their bioaccessibility is limited due to poor solubility in water and co-consuming dietary fat, especially in emulsified form, can improve carotenoids absorption. While dietary proteins have been studied as emulsifiers in oil-in-water emulsions, the influence of different protein sources on carotenoid bioaccessibility remains unclear. This study examined characteristics of six protein-based emulsifiers and assessed their effects on carotenoids bioaccessibility during in-vitro digestion.

Materials & Methods: Whey, casein, fish gelatin, soy, pea, and bovine gelatin have been prepared as emulsifiers in oilin-water emulsions and their mean particle size was analyzed. A carotenoids-rich salad was subjected to in-vitro digestion with each emulsion and ζ-potential and free fatty acid (FFA) release were measured to assess emulsion stability and lipid digestion.

Results & Discussion: Mean particle size for soy, bovine gelatin, casein and fish gelatin were 2.45 ± 0.08, 1.19 ± 0.01, 1.06 ± 0.01 and 2.05 ± 0.03 μm, respectively. Whey (0.77 ± 0.01 μm) showed the smallest size, while pea showed the largest size (32.98 ± 8.33 μm). After oral digestion, whey emulsion showed lower aggregation as indicated by ζpotential (-48.33 ± 6.86 mV), suggesting initial stability.

However, after intestine phase, soy (-50.02 ± 3.13 mV) and bovine gelatin (-50.22 ± 1.59 mV) emulsions exhibited significantly lower aggregation (p<0.05). Although statistically not significant, soy (73 ± 29 μL) and bovine gelatin (73 ± 15 μL) emulsions released higher amounts of FFA after the intestine phase, implying better lipid digestion and potentially greater carotenoids bioaccessibility.

Conclusions: Soy and bovine gelatin emulsions were stable throughout the digestion and showed better lipid digestion ability, supporting their potential for enhancing carotenoids delivery.

Practical Significance: Protein sources may play a critical role in carotenoids absorption when it is applied to the oilin-water emulsions.

Yueying YAO ¹, Nicole Min Yee WONG ¹, Jung Eun KIM ^1,2,*

¹Department of Food Science and Technology, Faculty of Science, National University of Singapore, Singapore. ²Bezos Center for Sustainable Protein, National University of Singapore, Singapore.

Background & Objectives: Angiogenesis is essential for vascular health since it induces recovery of blood vessels from endothelial dysfunction. Certain amino acids have gained attention for their beneficial effects in regulating endothelial function. Among them, L-arginine, abundant in plant-derived proteins such as legumes and nuts, and taurine, naturally occurring in animal-based foods like meat and fish, have particularly demonstrated vascular benefits. However, their roles in restoring angiogenic function under oxidative stress conditions remain insufficiently explored. This study investigated the angiogenic potential of L-arginine and taurine in human umbilical vein endothelial cells (HUVECs) pretreated with oxidized-LDL (ox-LDL).

Materials & Methods: HUVECs were pretreated with ox-LDL, followed by Larginine or taurine treatment. Cellular responses were assessed based on viability, intracellular reactive oxygen species (ROS) accumulation, nitric oxide (NO) production, and in vitro tube formation. The mRNA and protein expression levels of genes involved in the VEGFR2-PI3K/Akt/eNOS signaling pathway were also analyzed.

Results & Discussion: Findings revealed that both L-arginine and taurine significantly reduced ROS levels and recovered ox-LDL-induced impairment in angiogenesis, whereas only L-arginine increased NO production. Mechanistically, L-arginine activated the VEGFR2-eNOS/NO pathway, while taurine acted through VEGFR2- PI3K/Akt signaling.

Conclusions and: This research highlights the angiogenic capacity of both L-arginine and taurine under oxidative stress condition through their respective mechanisms. Further investigation is warranted to elucidate their precise mechanistic roles and clinical relevance in vascular pathophysiology.

Practical Significance: These findings suggest that L-arginine and taurine may serve as potential nutritional strategies to support endothelial repair and angiogenesis in conditions associated with oxidative stress and vascular dysfunction.

Marcus Ting¹, Ian En Kai Mak¹, Yueying Yao¹, Clarinda Nataria Sutanto¹, Zi Ning Leong¹, Chin Meng Khoo², Jung Eun Kim^1,3

¹Department of Food Science & Technology, National University of Singapore, Singapore
²Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
³Bezos Center for Sustainable Protein, National University of Singapore, Singapore

Email of presenter: marcusting@u.nus.edu

Background & Objectives
High protein diets (HPD) have gained popularity due to their health benefits like improving muscle protein synthesis hence, older adults are recommended to consume HPD. However, consuming HPD may increase blood advanced glycation end products (AGE) levels which are associated with chronic disease development. Therefore, this study aimed to investigate the effects of having HPD from different protein sources on blood and skin AGE levels in Singapore older adults.

Materials & Methods
This study was a 16-week randomized controlled trial and the control group consumed the My Healthy Plate (MHP) diet (n=19); the two intervention groups additionally consumed 20 g/d of casein (n=18), or soy (n=18) protein isolate with MHP diet. Plasma AGE levels, including N(6)-(1-carboxymethyl)-L-lysine (CML), N(6)-(1-carboxyethyl)-L-lysine (CEL), pentosidine and pyrraline, were quantified using ultra-performance liquid chromatography tandem triple quadrupole mass spectrometry at Week 0 and 16 and skin AGE level was measured via skin autofluorescence at Week 0, 8 and 16.

Results & Discussion
There were time and interaction effects for plasma CML levels (p_time=0.005, p_interaction=0.034) and specifically the soy group showed a significant increase (Week 0: 0.712±0.102, Week 16: 0.798±0.100, p-value=0.000). Plasma pentosidine levels was only significantly increased in the casein group (Week 0: 0.772±0.056, Week 16: 0.800±0.049, p-value=0.045). There was time effect for skin AGE level (p_time=0.045) and particularly the casein group showed significant increase (Week 0: 2.53±0.47, Week 8: 2.73±0.44, p-value=0.016). However, no significant changes were observed for plasma CEL and pyrraline levels regardless of time or group.

Conclusion and Practical Significance
HPD generally increased blood and skin AGE levels in Singapore older adults and different source of protein provides differential response in AGE levels. Present findings provided nutritional insights for shaping future guidelines for HPD in relation to AGEs management.

Presenting Author: Yu Lingyin¹
Email address: e1124962@u.nus.edu
Co-authors: Clarinda Nataria Sutanto¹, Ian Mak En Kai¹, Yao Yueying¹, Leong Zi Ning¹, Khoo Chin Meng², Jung Eun Kim^1,3

¹Department of Food Science and Technology, National University of Singapore, Singapore
²Division of Endocrinology, Department of Medicine, National University Hospital, Singapore
³Bezos Center for Sustainable Protein, National University of Singapore, Singapore

Background: Sleep is vital for overall health, while as individuals age, they experience poor sleep quality. Although healthy dietary patterns (HDP) and higher protein intake were suggested to improve sleep, these beneficial effects have not been well observed in Asian. Therefore, we aimed to assess the impact of different sources of higher protein intake when adhering to HDP on sleep quality in Singapore older adults.

Methods: This was a 16-week single-blind, parallel design study, and 55 participants (66 ± 4 yrs) were randomly assigned to the control (n = 19), casein (n = 18), or soy (n = 18) group. All groups followed HDP, while the casein and soy groups additionally consumed 20 g/d of respective protein isolate. Sleep quality was assessed by Pittsburgh Sleep Quality Index (PSQI) questionnaire to obtain sleep duration, sleep efficiency (SE), and sleep latency. Global sleep score (GSS) was calculated from PSQI, and good (GSS ≤ 5) and poor (GSS > 5) sleepers were also defined. Serum serotonin and urinary 6-sulfatoxymelatonin levels were measured.

Results: A significant time effect was detected on GSS (P_time=0.007) and SE (P_time=0.023) particularly in poor sleepers, and only soy group showed significantly decrease in GSS (week 0 vs week 8: 7.78 ± 1.93 a.u. vs 5.33 ± 1.63 a.u., P<0.01; week 0 vs week 16: 7.78 ± 1.93 a.u. vs 6.11 ± 1.59 a.u., P<0.05) and increase in SE (week 0 vs week 8: 76.83 ± 5.86 % vs 87.04 ± 7.86 %, P<0.05). No changes were found in other sleep components and biomarkers.

Conclusions and Practical Significance: Consuming higher protein with plant-based source along with adhering to HDP can enhance sleep quality, particularly in poor sleepers in Singapore older adults. This trial provides a feasible approach for Singaporean older adults to enhance sleep quality via dietary modification.

Kuoran Xing^1,2, Qiang Wang³, Narattam Sikdar¹, Yinglu Chen¹, Glebert Cañete Dadol¹, Ang Shi Jun^3,4,5, Jacqueline Tan^3*, David Tai Leong^1,2,6*

¹ Department of Chemical and Biomolecular Engineering, National University of Singapore
² NUS Graduate School of Integrative Science & Engineering Programme, National University of Singapore
³ A*STAR, Institute of High-Performance Computing (IHPC), Agency for Science, Technology and Research (A*STAR)
⁴ Singapore Integrative Biosystems and Engineering Research (SIBER), Agency for Science, Technology and Research (A*STAR)
⁵ Department of Chemistry, National University of Singapore
⁶ Bezos Center for Sustainable Protein at NUS

Mr. Kuoran Xing (xingkuoran@u.nus.edu) (Presenter)
A/P. David Leong (cheltwd@nus.edu.sg) (Lead PI)

Food safety assessment remains a critical public health priority, yet conventional multi-protocols often fall short in providing a decisive conclusion on the toxicity phenotype of many food-related compounds. Here, we present AI-Tox, the transcriptomic data-based AI framework for evaluating the toxicity of food substances. We generated a high-quality transcriptomic dataset by treating HepG2 and HepaRG liver cells with 32 chemicals (18 toxicants and 14 non-toxicants) at multiple concentrations, producing 195 expression profiles comprising close to 100 million raw reads. After rigorous features selection, we trained seven machine learning models with these large datasets and validated them via subset iterative processes, among which Linear Support Vector Machine achieving high predictive accuracy (precision up to 95%) for AI-Tox. The AI-Tox also identified key genes contributing to toxicity classification, and downstream gene ontology enrichment revealed cellular pathways related to mitochondria health, cytoskeleton disruption and apoptosis as key features in clustering. To assess novel food molecules, AI-Tox could perform accurate unsupervised prediction of their toxicity, validated by in vitro assays. AI-Tox demonstrates that transcriptomic profiling combined with powerful machine learning offers an accurate, sustainable, and mechanistically informed tool for food safety assessment.

F. Abiusia, F. Gao^a, C. Sägesser^a, B. Perez^a, A, Mathys^a.
ETH Zurich, Laboratory of Sustainable Food Processing, Zurich, Switzerland.
Presenter: Fabian Abiusi, fabian.abiusi@hest.ethz.ch

The polyextremophilic Galdieria sulphuraria is emerging as a promising microalgal species for food applications. G. sulphuraria can grow at pH values close to zero, temperatures up to 57°C, and in medium containing up to 2-3 M of salts. These unique characteristics can be leveraged to create a selective environment that prevents the proliferation of microbial contaminants, opening to continuous fermentation. G. sulphuraria has been cultivated using over 50 carbon sources, allowing for the combination of its cultivation with bioremediation of liquid residue streams from the food industry. This work explores the potential of heterotrophically cultivated G. sulphuraria as a protein producer for human consumption. To this end, the performances of four G. sulphuraria strains grown under the same conditions were compared, focusing on four aspects that make a strain suitable for protein production: 1) protein productivity, 2) amino acid profile, 3) protein bioaccessibility, and 4) color. Amino acid profiles varied among strains and growth phases, but all samples met FAO dietary requirements for adults. The specific growth rates were between 1.06 and 1.48 day-1. After glucose depletion, all strains showed an increase of 38-49% in nitrogen content within 48 hours, reaching 7.8-12.0% w/w. An opposite trend was observed in protein bioaccessibility, which decreased on average ABSTRACT SUBMISSION from 69% during the exponential phase to a minimum of 32% 48 hours after the stationary phase, with significant differences among the strains. Two of the tested strains displayed low pigmentation when cultivated heterotrophically, while the other two maintained their pigmentation. Therefore, selecting the appropriate strain and harvesting time is crucial for successful single-cell protein production.

Shaymaviswanathan Karnaneedi^1,2,3‡, Elecia B. Johnston^1,2,3‡, Thimo Ruethers^1,2,3,4, Utpal Bose⁵, Angéla Juhász⁶, James A. Broadbent⁵, Emily Jerry^1,2,3, Sally Stockwell⁵, Keren Byrne⁵, Sandip D. Kamath^1,2,3, Vachiranee Limviphuvadh^7,8, Sebastian Maurer-Stroh^7,8,9, Michelle L. Colgrave^5,6, Andreas L. Lopata^{1,2,3,4* *}andreas.lopata@jcu.edu.au

¹ Molecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Townsville, Queensland, Australia;
² Australian Institute of Tropical Health and Medicine, James Cook University, Townsville, QLD, Australia;
³ Centre for Food and Allergy Research, Murdoch Children’s Research Institute, Melbourne, VIC, Australia;
⁴ Tropical Futures Institute, James Cook University Singapore, Singapore;
⁵ CSIRO Agriculture and Food, 306 Carmody Rd, St Lucia QLD 4067, Australia;
⁶ Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, School of Science, Edith Cowan University, Joondalup WA 6027, Australia;
⁷ Biomolecular Function Discovery Division, Bioinformatics Institute, Agency for Science, Technology and Research, Singapore;
⁸ IFCS Programme, Singapore Institute for Food and Biotechnology Innovation, Agency for Science, Technology and Research, Singapore;
⁹ Department of Biological Sciences, National University of Singapore, Singapore. ‡Equally contributing authors

Background & Objectives
As the global population grows, edible insects are emerging as a sustainable protein source for both human consumption and animal feed. However, emerging evidence highlights the presence of allergenic proteins in insects, particularly due to cross-reactivity with known crustacean allergens. While tropomyosin remains the most studied insect allergen, the broader allergen profile and the influence of protein extraction methods on immunogenicity remain underexplored. This study investigates the allergen profiles of edible cricket (food) and black soldier fly larvae (feed) products.

Materials & Methods
Crude protein extracts were generated from Acheta domesticus (cricket) and Hermetia illucens (black soldier fly; BSF) food/feed products using five different protein extraction methods. Immunoblotting with sera from individuals with confirmed crustacean allergy revealed IgEbinding proteins, identified through LC-MS and a custombuilt insect proteome databases. Cricket-based food products manufactured for human consumption, including whole cricket snacks, corn chips, and energy bars, and BSF-based pet food products were tested with different commercial test kits.

Results & Discussion
Proteomic analysis identified six IgE-binding proteins in cricket extracts: tropomyosin, myosin light chain, vitellogenin, troponin-I, and apolipophorin-III. BSF extracts revealed seven IgE-binding proteins, including hemocyanin, tropomyosin, ATP-synthase, HSP20, and odorant/chitin-binding proteins. Notably, a major crustacean allergen, was absent in all insect preparations. Quantitative proteomics revealed multiple isoforms of crustacean allergen homologues in crickets, influenced by extraction method.

Conclusions
Our findings identify several novel cross-reactive insect allergens and highlight significant variability in allergen detection based on processing and preparation methods. These insights have critical implications for allergy risk assessments. Accurate characterization of these allergens is essential to support regulatory frameworks, inform consumer safety, and guide the development of hypoallergenic insect-based products.

Practical Significance
The urgent development of insect-specific allergen detection tools is essential to ensure accurate food labelling, build consumer trust, and enable the safe and sustainable expansion of insect-based food and feed industries.

Jorry Dharmawan, Tan Su Hui and Susanna Leong Su Jan

Singapore Institute of Technology
1 Punggol Coast Road, Singapore 828608

Email: jorry.dharmawan@singaporetech.edu.sg

Pea protein has gained prominence as a viable alternative protein source due to its nutritional value and sustainability. However, its commercial utilization is hindered by undesirable flavours that affect consumer acceptance negatively. To address this issue, enzymatic and heat treatments were employed to mitigate off-flavours in pea protein. This study aims to investigate the impact of those techniques on the flavour profile of pea protein isolate (PPI) and pea protein concentrate (PPC). Enzymatic treatment was conducted using protein glutaminase, while thermal treatment was performed at 70°C for 1 hour. Sensory evaluation of both untreated and treated samples was carried out by 12 trained panellists using descriptive analysis method to assess the efficacy of the treatments in reducing off-flavours.

The results show that enzymatic treatment significantly altered the flavour profile for both PPI and PPC. Enzymatically-treated PPI showed a reduction in the beany note, accompanied by an increase in saltiness and umami compared to the untreated PPI. Conversely, enzymatically-treated PPC was less green and bitter, while exhibiting an increase in beany, salty, and astringent characteristics relative to untreated PPC. Similarly, the flavour profile of PPC was modified following heat treatment with a notable reduction in undesirable green notes and bitterness, rendering it more palatable than both untreated and enzymatically-treated PPC. In contrast, heat-treated PPI showed minimal improvement, with a slight increase in beany and cooked notes post-treatment. This could be attributed to the initial heating that PPI underwent during processing, resulting in negligible changes to its flavour during subsequent thermal treatment. In conclusion, enzymatic treatment proves to be an effective method for mitigating off-flavours in PPC and PPI, while heat treatment enhances the flavour of PPC, thereby improving consumer acceptance and supporting the sustained growth of plant-based foods.

(Jason) Xing KANG¹, Jenny GOH¹, Levene W. CHUA¹, Han XIAO¹, Sunny H. WONG¹, Joseph J.Y. SUNG¹, Yusuf ALI¹

¹Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore
jason.kangx@ntu.edu.sg

Background & Objectives:
The rising popularity of plantbased proteins has drawn attention to their impact on health impacts. This study compares the effects of plant-derived (soybean) and animal-derived (chicken) proteins on gut health, metabolism, and inflammation in aging mice.

Materials & Methods:
Aged C57BL/6 mice (64 weeks old) were fed diets enriched with either soybean (AS) or chicken (AC) protein (20% of calories from protein) for 16 weeks. Glucose metabolism (oral glucose tolerance tests, OGTT), gut microbiome diversity (metagenomic sequencing), intestinal permeability (serum lipopolysaccharides [LPS]), inflammation (cytokine profiling and immune cell analysis), and tissue-level changes (liver, colon, adipose histology) were assessed. Nutritional peptide profiles were analyzed via peptidomics.

Results & Discussion:
The different diets resulted in changes in food intake, body weight gain, and accumulation of subcutaneous and visceral fat. Liver histology showed increased lipid droplet deposition. Impaired glucose tolerance (higher OGTT AUC) and elevated serum LPS levels indicated compromised gut barrier integrity. AC-fed mice also exhibited splenomegaly ABSTRACT SUBMISSION and heightened inflammation, highlighted by increased TNF-α⁺ CD8⁺ T cells in spleen and PBMC, and higher cytokine levels in blood. Similarly, gut microbiome analysis showed distinct compositional differences correlating with the metabolic and inflammatory outcomes.

Conclusions:
Different protein sources alter host effects related to metabolic dysfunction, inflammation, and gut barrier physiology. These findings highlight the potential of plant-based proteins to promote health.

Practical Significance:
This study demonstrates that plant-based proteins, like soybean, may promote healthier aging by improving glucose metabolism, reducing inflammation, and supporting gut health. Incorporating plant-derived proteins into diets could be a practical dietary strategy to counteract age-related health issues, particularly in elderly populations.

Sachini Upeka Perera^1,2>, Hena Abdu^1,2, Shahna Manakat Vakeri^1,2, Mohamed Salim^1,2, Zubeen Hathi^1,2, Blaise Leopold Tardy^1,2, Srinivas Mettu^*1,2
Presenter Email: srinivas.mettu@ku.ac.ae

1. Department of Chemical and Petroleum Engineering, Khalifa University, United Arab Emirates
2. Food Security and Technology Center (FSTC), Khalifa University, Abu Dhabi, United Arab Emirates

Conventional dairy (milk) production uses significant amounts of land and water, emitting huge amounts of greenhouse gases. Sustainable production of cow-less milk through the route of mycelium (fungal) and bacterial fermentation is highly needed. We address this issue by unique approach of harnessing symbiotic filamentous fungi with human gut microbes to produce proteins and fats together in one bioreactor system.

Mycelium, the thread-like structure called hyphae, also known as the vegetative part of fungi, has already proven to be a promising sustainable alternative to traditional materials. In this study, we investigate the structural features, and multifunctional potential of mycelial hydrogels derived from three fungal species: Trichoderma harzianum, Ganoderma lucidum, and Pleurotus ostreatus. The fungi were cultivated in liquid culture under controlled environmental conditions to facilitate consistent hydrogel formation for a period of 3-4 weeks.

Microstructural characteristics of these mycelium hydrogels were analyzed using Scanning Electron Microscopy (SEM) after separation from liquid culture and freeze drying, revealing differences in filament arrangement and network architecture across species. Water content and density analyses were conducted to assess water uptake and hydration properties.

Mechanical testing, including compressive and tensile strength measurements, was performed to evaluate the structural integrity and flexibility of the hydrogels, critical for material performance. Chemical and crystallographic properties of the dried samples were further explored using Fourier Transform Infrared Spectroscopy (FTIR) and X-ray Diffraction (XRD). These analyses provide insights into molecular bonding patterns and the degree of crystallinity within the mycelial matrices.

A novel aspect of this research is the immobilization of probiotic anaerobic bacteria within the hydrogel networks, aiming to assess potential symbiotic interactions and the feasibility of using mycelium as a living scaffold. This approach provides a platform for further studies on the application of mycelium as living materials in engineered bio-processing technologies.

Ziqi Li^a (zili5377@uni.sydney.edu.au), Valeria Messina^b, Daniel J. Skylas^c, Peter Valtchev^b, Chris Whiteway^c, Brent N. Kaiser^b, Fariba Dehghani^a.

^a School of Chemical and Biomolecular Engineering, The University of Sydney, Camperdown, NSW, Australia
^b School of Life and Environmental Sciences (SOLES), The University of Sydney
^c Australian Export Grains Innovation Centre, North Ryde, NSW, Australia

Background & Objectives
Soy protein has long dominated the plant-based protein market due to its high nutritional and functional values. However, its allergenicity and environmental impact are driving interest in alternative pulses such as faba bean and yellow pea. This study aims to evaluate the protein quality and functional properties of Australian faba bean and yellow pea protein concentrates and isolates, as well as comparison with traditional soy proteins.

Materials & Methods
Faba bean (FB), yellow pea (YP) and soy (S) protein concentrates (PC) and isolates (PI) were obtained by dry and wet fractionation techniques. Amino acid composition and in-vitro protein digestibility were assessed to determine protein quality. Secondary structure was analysed using Fourier-transform infrared spectroscopy (FTIR). Key functional properties were evaluated (waterholding capacity (WHC), protein solubility, and foaming properties).

Results & Discussion
Amino acid analysis revealed that FBPC, FBPI, YPPC and YPPI showed deficient in sulfur-containing amino acids, a similar trend was observed for SPI and SPC. In-vitro protein digestibility showed competitive levels for FBPC (84.5%), FBPI (92.5%), YPPC (83.7%), and YPPI (94.1%) when compared to SPC (87.5%) and SPI (95.0%). Significant differences (p<0.05) were observed for secondary structure between PC and PI for FB, YP and S, especially in the amide I band. FBPC and YPPC exhibited greater solubility (46.2% and 50.1%, respectively) when compared to SPC (42.5%). On the other hand, FBPI and YPPI showed higher WHC (4.3 g/g and 4.0 g/g, respectively) versus SPI (3.8 g/g) and SPC (3.2 g/g).

Conclusions and Significance
FB and YP ingredients (PC and PI) generally exhibited higher functional properties compared to SPI and SPC, highlighting their potential to offer food manufacturers more diverse and flexible options for tailoring texture, stability, and nutritional profiles in both new and reformulated plant-based products.

Hopf Andreas^1*, Agarwal Deepa², Skylas Daniel J.³, Whiteway Chris³, Valtchev Peter¹, Kaiser Brent N.⁴, Buckow Roman^1,2 and Dehghani Fariba¹

E-mail of corresponding author
andreas.hopf@sydney.edu.au

¹ School of Chemical and Biomolecular Engineering, University of Sydney, Darlington, NSW, Australia
² La Trobe Institute for Sustainable Agriculture & Food (LISAF), School of Agriculture, Biomedicine and Environment, La Trobe University, Bundoora, VIC, Australia
³Australian Export Grains Innovation Centre, North Ryde, NSW, Australia
⁴School of Life and Environmental Sciences, Faculty of Science, University of Sydney, Camperdown, NSW 2006, Australia

Dry-fractionated faba bean protein concentrates (FPCs) offer promising applications in the food industry, particularly in the formulation of plant-based meat and dairy alternatives, because of their versatile functional characteristics. This study examined the impact of thermal treatment on functional properties of FPC and the textural properties of its gels when using CaSO4 as a coagulant.

The thermal treatment consisted of a saturated steam treatment followed by fluidized-bed drying, applied to dehulled faba bean seed material prior to dry fractionation. The functional properties of untreated and thermally treated FPCs, including protein solubility, water-holding capacity, and thermal properties were assessed. Subsequently, heatinduced gels were prepared with FPC concentrations ranging from 12% to 16% w/w, and CaSO4 concentrations from 0.0% to 0.5% w/w, respectively.

The gels were analyzed for water-holding capacity, pH, textural and rheological properties. Thermal treatment resulted in a significant reduction (-32%) of protein, while enhancing water-holding capacity of the FPC material by 2.2-fold. Gels formed with thermally treated FPC exhibited significantly higher stiffness and hardness, as well as altered rheological properties, including higher storage, loss modulus, yield stress and flow stress. The combination of thermal treatment and CaSO4 significantly improved the textural and rheological properties of the gels, suggesting a synergistic effect between pre-treatment and the salt coagulant.

The results of this study were applied to design a prototype soy-free tofu from faba beans with improved gel structure. These findings demonstrate the potential of thermal treatment and salt coagulants to modulate the gelation properties of pulse protein concentrates, offering sustainable strategies for developing plant-based edible gels with enhanced functionality.

List of authors:

1. Feizya Hilyata Musaddiq
2. Inaya Chairani Mahdiya Putri

Email contact:
feizyamusaddiq@gmail.com

Tempeh, commonly made from soybean, has been a staple source of protein in Indonesia for more than 300 years. Due to the import taxes, soybean tempeh is considered a nonsustainable food source, since Indonesia is still dependent with 80% soybean imports to meet the national needs. Substitution of soybean in tempeh is a solution to lowering the dependency.

This study aimed to develop and mimicking alternative tempeh with higher protein content by identifying formulation, F0 value, and sensory profile tempeh made from jackbean, mungbean and okara packed in a retort pouch, using hedonic ranking, CATA (Check-All-That-Apply) and RATA (Rate-All-That-Apply) test.

The Fermentation of tempeh using Rhizopus oligosporus. F0 value calculated using general methods with heat sufficiency validation test using thermocouple and datalongger resulting in F0 value 7.4 minutes. Each panellist given a set of trays containing 3 sample formulations for hedonic, continued by CATA and RATA to determine consumer perceptions of the presence characteristics, through a tick mark scale for CATA and identification level scale for RATA (i.e. 1: Not noticeable to 6: very noticeable).

Formulation two named F2 (50% jackbean, 30% mungbean and 20% okara) are chosen by the panellist. F2 is similar to soybean tempeh with higher protein content and variety of essential amino acids and has the 96.8% of desire attribute from ideal tempeh with no significant differences attribute, interpreted as F2 successfully mimicking commercial products. F0 value 7.4 minutes resulting in a shelf-life stable product. These Finding of Ready-to-Eat Tempeh from Jackbean and Mungbean Enriched with Okara can be a solution for soybean import tax, reduce 30.46% soybean consumption (equal to 13,000 ton/year), manage 12.5% waste from utilization 800,000kg/year okara, contribute to affordable functional food, diversifying legume use in tempeh production, enhancing food accessibility.

Keywords: Tempeh, Jackbean, Mungbean, Okara