Cultured meat technology holds great promise and can help address the sustainability and environmental challenges faced by traditional livestock husbandry. Unlike other alternative proteins， cultured meat is made from animal stem cells cultured in vitro and is the closest product to natural meat. To make cultured meat have the sensory and nutritional characteristics of conventional meat as much as possible， researchers have conducted extensive studies on scaffold characteristics and manufacturing techniques， with the aim of replicating the properties of natural muscle tissue. This review summarizes the methods for preparing edible scaffolds， with the aim of producing tissue-like cultured meat with aligned myotubes. Firstly， the composition and structure of animal-derived meat are analyzed. And， based on the structural properties of natural muscle tissue， the main considerations for the design of scaffolds for cultured meat engineering are elaborated， with the formation of uniaxially aligned myotubes as the key focus. Secondly， the main scaffold techniques suitable for generating aligned myotubes and their current applications in cultured meat engineering are summarized. Finally， the future development prospects are outlined.

Objective: To study the expression levels of differentially expressed genes in Candida parapsilosis under high glucose stress and to investigate its stress response mechanism. Methods: The yeast presented in the contaminated condensed milk during storage were isolated and characterized by plate screening and 18s DNA sequencing， and the total RNA of the strains was extracted to construct cDNA libraries. Differentially expressed genes were analyzed by transcriptome analysis and verified by real-time fluorescence quantitative PCR. Results: The yeast was identified as Candida parapsilosis， its optimal temperature and pH value were 35 ℃ and 8， respectively. It could still grow slowly when the glucose concentration reached 50%. The main product in the fermentation broth was sorbitol， with a yield of up to 41.06 g/L. A total of 2 066 significant differential genes were detected in the transcriptomic analysis， of which 993 were significantly up-regulated and 1 073 were significantly down-regulated. The genes related to heat stress proteins (Hsp70， Hsp90)， amino acid transport， glycogen transport， glycolysis and ethanol dehydrogenase were significantly down-regulated， while the genes related to amino acid permease， glycogen synthesis， and synthesis of certain amino acids (e.g.， glutamate， proline， arginine， cysteine， etc.) with osmotic stress-protecting effects were significantly up-regulated. Conclusion: Candida parapsilosis could improve performance under hyperosmotic stress by synthesizing certain sugars or amino acids (e.g.， proline， glycine， and glutamate).

Objectives: Glyphosate was an efficient herbicide， while there was negative impact on water environment and human health. Therefore， it is significant to develop an efficient and accurate method to detect the glyphosate in water. In this study， a fluorescent turn-on probe was designed based on R-phycoerythrin in Porphyra haitanensis for detection of glyphosate in water. Methods: The crude extract in P. haitanensis was extracted by freeze-thaw cycle and ultrasonication. The peptide mass fingerprint， abosorbance spectrum and SDS-PAGE indicated the extract was R-phycoerythrin. The purity of R-phycoerythrin was as high as 4.96 after purification. Further studies indicated the R-phycoerythrin was specific fluorescence responsed to Cu2+ ions. A fluorescent probe based on the copper ion quenching R-phycoerythrin fluorescence was designed. Results: The probe fluorescence signal showed good linear relationship with the glyphosate concentration， and the detection limit was 443.4 μg/L， which was significantly lower than the maximum allowable residue limit (0.7 mg/L) specified for glyphosate in the Chinese National Standard for Drinking Water Quality (GB 5749—2022). The probe was used to detect the glyphosate content in two water samples and the glyphosate was tested. Conclusions: The study not only highly-value utilized of P. haitanensis， but also provides a new method for the detection of glyphosate residues in water， which has certain scientific significance and application value.

Volatile metabolites of Clavispora lusitaniae J5 in different culture were studied. The extracts of potatoes， sorghum， wheat， and red millet were used as the culture medium to cultivate Clavispora lusitaniae J5. The volatile metabolites were investigated by means of headspace solid-phase microextraction-two dimensional gas chromatography quadrupole time-of-flight mass spectrometry， and analyzed by chemometric methods. A total of 65 volatile compounds were detected in cultures of Clavispora lusitaniae J5. There was greater abundance of volatile components in HXM， and greater total concentration of volatile components in XM. There was no significant difference in the concentration of ethanol， ethyl hexanoate， ethyl octanoate， phenylethyl acetate， acetaldehyde and ethyl heptanoate among the three types of grain cultures. 40 Key volatile metabolites with variable importance in projection (VIP)>1 and significant differences in content were obtained by the orthogonal partial least-squares regression discriminant analysis model， as well as 5 volatile metabolites with significant differences in content with VIP<1. Hexanol， butyl acetate， heptanal， 3-methylbutanal， 2-pentyl furan， 1-octen-3-ol， 3-methyl-1-butanol， cis-2-heptanal were the important aroma compounds with relative odor activity values (ROAV)>1 in the culture of HXM. Although they were different from the important aroma compounds in the culture of XM， they were common aroma components in fermented wine. Therefore， it is feasible to cultivate Clavispora lusitaniae J5 with red millet for enhancing the aroma of the red-millet Huangjiu.

Objective: To explore the effect of Na+/K+ in the intestinal environment on the interaction between soy hull polysaccharide (SHP) and intestinal mucus， and its effect on the diversity of intestinal flora. Methods: Molecular docking and molecular dynamics simulation were used to analyze the main binding force and complex stability of the target protein and soybean seed coat polysaccharide after Na+/K+ treatment. The effects of SHP and mucin on the changes of OTU and species abundance of intestinal flora after Na+ treatment was detected by 16S rDNA genomics. Results: The main force of SHP binding to mucin after Na+/K+ treatment was hydrogen bond. Within 100 ns， the RMSD value， RSMF value， Rg value and the number of hydrogen bonds of the complex structure fluctuated slightly. The secondary structure β-sheet of mucin mainly existed in anti-parallel mode after Na+ treatment， while the secondary structure β-sheet of mucinmainly existed in parallel mode after K+ treatment. The binding free energy [(196.7±26.7) kJ/mol] and the contribution of amino acid residues (18.18 kJ/mol) of SHP to mucin after Na+ treatment were greater than those after K+ treatment [(158.5±26.7) kJ/mol and 10.63 kJ/mol， respectively]. In the presence of Na+， the diversity of intestinal flora was up to 284 OTUs when SHP was used as a substrate for 8 h， and the abundance of various dietary fiber degrading bacteria was increased， and the abundance of Escherichia coli-Shigella was decreased. At the same time， the abundance of Bifidobacterium and Lactobacillus was also decreased. Conclusion: After Na+ treatment， the binding ability and stability of SHP and mucin are higher， and the diversity and abundance of intestinal flora are affected. Na+ treatment has a potential effect on the whole intestinal function of dietary fiber.

As a new type of protein resource that does not rely on traditional livestock or fishery production， NeoProtein is a core strategic direction for practicing the ｀Big Food Concept' and safeguarding national food supply security. It holds great significance for promoting the upgrading of the food industry and realizing global sustainable development. Although the NeoProtein industry is currently showing a trend of rapid development， it still faces many bottlenecks in aspects such as technology industrialization and product quality optimization. Based on the current development status of the industry and the 2025 technology trends， this paper sorts out and identifies the top 10 technical challenges in the field of NeoProtein， and systematically expounds the background， challenges and solutions for each challenge. It is expected to provide references for scientific research， industrial innovation and policy formulation in related fields， and contribute to the high-quality development of the NeoProtein industry.

Citrus is a globally significant economic crop with the largest cultivation area and highest production yield. Currently， its processed products are still dominated by juice， canned goods， and similar items， resulting in relatively low comprehensive utilization of citrus resources. Citrus peels， pomace， physiological fruit drops， and other by-products are rich in high-value bioactive components such as pectin， flavonoids， and essential oils， which have wide applications in food， pharmaceuticals， chemical engineering， and other fields. This article reviews the status and technological bottlenecks in the citrus processing industry， systematically outlines the properties， biological activities， and advancements and limitations of traditional extraction technologies for key functional components in citrus resources. It also summarizes the technical advantages of synthetic biology in the green manufacturing of natural products， with a focus on recent progress in microbial biosynthesis of citrus flavonoids. In line with the demands for resource recycling and green， low-carbon development， synthetic biology is expected to open new pathways for obtaining high-value active ingredients from citrus， thereby promoting the industry's transition toward higher added value and sustainable development.

Objectives: The efficient release of bioactive peptides (BPs) from bovine milk proteins is of great significance. This study aimed to explore the potential of using the soluble proteinase of lactic acid bacteria (LAB) for production of BPs. Methods: A LAB strain exhibiting high proteolytic activity was screened and identified. The gene encoding the proteinase in the selected LAB was amplified via PCR and then modified by removing the sequence encoding the cell-wall-associated region， enabling it to be a soluble protein. Subsequently， the modified gene prtB3 was cloned and expressed in the food-grade expression strain Lactococcus lactis NZ9000. After purification， a comprehensive evaluation was conducted on the enzymatic properties of the recombinant proteinase PrtB3 and the biological activities of the hydrolysates resulting from casein degradation. Results: The screened LAB strain was identified as Lactobacillus delbrueckii subsp. lactis. A soluble， secreted， C-terminally Myc and His-tagged derivative PrtB3 was constructed and expressed in Lactococcus lactis via the NICE■ Expression System. Compared with the wild-type proteinase， PrtB3 demonstrated a significant expansion in substrate specificity. It was proteolytically active in the hydrolysis of α-， β-， and κ-casein. Regarding biological activity， the casein hydrolysate produced by PrtB3 showed remarkable angiotensin-converting enzyme (ACE) inhibitory activity. The maximum ACE inhibition rate reached 78%， which was twice that of the hydrolysate produced by the wild-type proteinase. The half-maximal inhibitory concentration (IC50) was 1.69 mg/mL， and the ACE inhibitory activity remained stable over an 8-week period. Furthermore， the enzymatically produced hydrolysate showed antimicrobial activity against Staphylococcus aureus and Escherichia coli. Conclusions: In this study， the soluble proteinase PrtB3 was successfully cloned and expressed. It effectively enhanced the degradation of casein and significantly improved the biological activity and stability of the casein hydrolysate. PrtB3 demonstrated great potential in producing BPs by degrading casein， providing a novel approach for the preparation of bioactive peptides.

This study aimed to systematically investigate the dynamics of microbial community structure and volatile flavor compounds during the long-term storage of Citrus medical L. var. sarcodatylis and to elucidate their interrelationships. Microbial communities in samples stored for 4， 7， 10， and 12 years were analyzed using high-throughput sequencing， while volatile components were identified by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). At the phylum level， all samples were predominantly composed of Firmicutes， Proteobacteria， Bacteroidetes， and Actinobacteria. Analysis of volatile compounds revealed 9 chemical categories encompassing 287 key flavor substances， samples from 2014 were rich in ethers (~35%) and hydrocarbons (~30%)； those from 2017 showed a marked increase in alkenes (~40%)； by 2020， hydrocarbon content declined to ~12%， whereas esters and alcohols increased significantly. In conclusion， although the microbial community remained relatively stable throughout storage， the volatile flavor profile underwent considerable transformation， and these shifts were closely associated with microbial succession. These findings provide insights into the flavor development of Citrus medical L. var. sarcodatylis from a microbial-metabolic perspective and offer a scientific basis for optimizing its production and quality control.

Exploring the inhibitory effect of small molecules on pancreatic lipase (PL) has become an important method for screening active ingredients with anti-obesity effect in vitro. The binding reaction and mechanism of genistein (Gen) and naringenin (Nar) with PL were studied by fluorescence spectroscopy， non-radiative energy transfer theory and high performance liquid chromatography. At the same time， three-dimensional fluorescence， surface hydrophobicity， X-ray diffraction， infrared spectroscopy， particle size， differential scanning calorimetry， and scanning electron microscopy were used to characterize the structure of the composites from different angles， and their influence on the structure and properties of PL were comprehensively clarified. The results showed that the quenching of PL by Gen and Nar was static quenching， and the binding constants were 42.39 L/mg and 22.57 L/mg， respectively. The addition of both reduced the α-helix content of PL， increased the content of β-turn， β-sheet and random coil， and increased the crystallinity from 25.60% to 32.77% and 38.00%， respectively. The surface hydrophobicity decreased and the thermal stability decreased (the denaturation temperature decreased from 76.04 ℃ to 65.00 ℃ and 65.05 ℃， respectively)， and the PL changed from a smooth sheet structure to a rod-like particle. These results indicate that Gen and Nar significantly affect the structure and properties of PL， providing new ideas for the development of new PL inhibitors.