Pub Date : 2026-01-29DOI: 10.3390/fermentation12020074
Han Zhang, Xu Tang, Yaping Xu, Shen Yang, Peng Wu
This study investigated the generation of α-glucosidase inhibitory peptides from whey protein fermented by the marine-derived probiotic Lacticaseibacillus casei DS31 (isolated from the intestinal microbiota of the large yellow croaker, Larimichthys crocea) and assessed their potential for practical glycemic management. Fermentation markedly increased inhibitory activity, with the freeze-dried crude supernatant exhibiting an IC50 of 2.115 mg/mL. Activity was further enriched through stepwise purification: ultrafiltration (<3 kDa) improved potency (IC50 = 1.206 mg/mL), and subsequent Sephadex (crosslinked dextran) G-15 gel filtration yielded a more active E fraction (IC50 = 1.145 mg/mL). LC–MS/MS characterized 19 peptides, and integrated in silico screening (PeptideRanker combined with molecular docking) highlighted GEPGPEGPAG as a leading candidate, showing a more favorable predicted binding energy (−82.50 kcal/mol) than the positive control acarbose (−69.31 kcal/mol). Docking analysis suggests that GEPGPEGPAG may inhibit α-glucosidase by forming a stable network of hydrogen bonds, salt bridges, and hydrophobic interactions within the catalytic pocket. Overall, DS31-fermented whey and its enriched fractions show promise as functional ingredients for postprandial glycemic control.
{"title":"Fermentation-Driven Generation of α-Glucosidase Inhibitory Whey Peptides by Marine-Derived Probiotic Lacticaseibacillus casei DS31: Activity Enrichment and Peptidomics","authors":"Han Zhang, Xu Tang, Yaping Xu, Shen Yang, Peng Wu","doi":"10.3390/fermentation12020074","DOIUrl":"https://doi.org/10.3390/fermentation12020074","url":null,"abstract":"This study investigated the generation of α-glucosidase inhibitory peptides from whey protein fermented by the marine-derived probiotic Lacticaseibacillus casei DS31 (isolated from the intestinal microbiota of the large yellow croaker, Larimichthys crocea) and assessed their potential for practical glycemic management. Fermentation markedly increased inhibitory activity, with the freeze-dried crude supernatant exhibiting an IC50 of 2.115 mg/mL. Activity was further enriched through stepwise purification: ultrafiltration (<3 kDa) improved potency (IC50 = 1.206 mg/mL), and subsequent Sephadex (crosslinked dextran) G-15 gel filtration yielded a more active E fraction (IC50 = 1.145 mg/mL). LC–MS/MS characterized 19 peptides, and integrated in silico screening (PeptideRanker combined with molecular docking) highlighted GEPGPEGPAG as a leading candidate, showing a more favorable predicted binding energy (−82.50 kcal/mol) than the positive control acarbose (−69.31 kcal/mol). Docking analysis suggests that GEPGPEGPAG may inhibit α-glucosidase by forming a stable network of hydrogen bonds, salt bridges, and hydrophobic interactions within the catalytic pocket. Overall, DS31-fermented whey and its enriched fractions show promise as functional ingredients for postprandial glycemic control.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"12 2","pages":"74-74"},"PeriodicalIF":0.0,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/12/2/74/pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147381629","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-05DOI: 10.3390/fermentation12010028
Xian Tao, H. L. Li, Jie Zeng, Mei Fang Peng, Qing Liu, Lan Luo, Yan Wang, Juan Yang, X L Yang, Liangqun LI
Alcoholic liver injury (ALI) is a major global public health issue, with oxidative stress imbalance as its core pathological mechanism. The Kelch-like ECH-associated protein 1–nuclear factor erythroid 2-related factor 2–heme oxygenase-1/glutathione peroxidase 4 signaling pathway (Keap1–Nrf2–HO-1/GPX4) signaling pathway is a key target for regulating hepatic antioxidant defense. This study integrated Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS), Global Natural Products Social Molecular Networking (GNPS) molecular networking, network pharmacology, and animal experiments to systematically explore the hepatoprotective effect and mechanism of Cornus officinalis yeast-fermentation (COF). Component characterization identified 25 bioactive components, including flavonoids, triterpenic acids, and other fermentation-derived metabolites. Network pharmacology identified 441 common targets and 36 core targets of COF and ALI, which were enriched in oxidative stress regulation, inflammatory response, and the Keap1–Nrf2 pathway via Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Molecular docking showed that icariin and other components had stable interactions with Keap1 and Nrf2 (binding energy < −5 kcal/mol). Animal experiments confirmed that COF reduced the liver index of ALI mice, downregulated serum Alanine Aminotransferase (ALT)/Aspartate Aminotransferase (AST) activities, and ameliorated liver pathological damage. Western blot verified that COF inhibited Keap1 expression, promoted Nrf2 nuclear translocation, and upregulated HO-1/GPX4 expression. In conclusion, COF alleviates hepatic oxidative stress by regulating the Keap1–Nrf2–HO-1/GPX4 pathway, providing a scientific basis for its development as a functional food or candidate drug against ALI and a technical paradigm for fermentation-enhanced medicinal plant research.
{"title":"Fermented Cornus officinalis Fruit Protects Against Alcohol-Induced Liver Injury via Regulating Keap1–Nrf2–HO-1/GPX4 Pathway: UPLC-MS/MS Characterization, Network Pharmacology, and Animal Validation","authors":"Xian Tao, H. L. Li, Jie Zeng, Mei Fang Peng, Qing Liu, Lan Luo, Yan Wang, Juan Yang, X L Yang, Liangqun LI","doi":"10.3390/fermentation12010028","DOIUrl":"https://doi.org/10.3390/fermentation12010028","url":null,"abstract":"Alcoholic liver injury (ALI) is a major global public health issue, with oxidative stress imbalance as its core pathological mechanism. The Kelch-like ECH-associated protein 1–nuclear factor erythroid 2-related factor 2–heme oxygenase-1/glutathione peroxidase 4 signaling pathway (Keap1–Nrf2–HO-1/GPX4) signaling pathway is a key target for regulating hepatic antioxidant defense. This study integrated Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS), Global Natural Products Social Molecular Networking (GNPS) molecular networking, network pharmacology, and animal experiments to systematically explore the hepatoprotective effect and mechanism of Cornus officinalis yeast-fermentation (COF). Component characterization identified 25 bioactive components, including flavonoids, triterpenic acids, and other fermentation-derived metabolites. Network pharmacology identified 441 common targets and 36 core targets of COF and ALI, which were enriched in oxidative stress regulation, inflammatory response, and the Keap1–Nrf2 pathway via Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Molecular docking showed that icariin and other components had stable interactions with Keap1 and Nrf2 (binding energy < −5 kcal/mol). Animal experiments confirmed that COF reduced the liver index of ALI mice, downregulated serum Alanine Aminotransferase (ALT)/Aspartate Aminotransferase (AST) activities, and ameliorated liver pathological damage. Western blot verified that COF inhibited Keap1 expression, promoted Nrf2 nuclear translocation, and upregulated HO-1/GPX4 expression. In conclusion, COF alleviates hepatic oxidative stress by regulating the Keap1–Nrf2–HO-1/GPX4 pathway, providing a scientific basis for its development as a functional food or candidate drug against ALI and a technical paradigm for fermentation-enhanced medicinal plant research.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"12 1","pages":"28-28"},"PeriodicalIF":0.0,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147381472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-03DOI: 10.3390/fermentation12010024
Fenghao An, Yanchun Feng, Da Li, Mei Hua, Xiuquan Wang, Xifei Xu, Yuguang He, Xinyu Miao, Mubai Sun, Honghong Niu, Hongyan Xu, Jinghui Wang
As one of the few carotenoid-producing microorganisms, Rhodotorula mucilaginosa remains underexplored for its antioxidant activity. This study investigated the effects of R. mucilaginosa JAASSRY on D-galactose-induced aging mice. The high-dose JAASSRY (HR) significantly increased body weight by 9.89% compared to the model group (AM), while reducing organ indices of the spleen, liver, kidneys, and brain (p < 0.01). Compared with the AM group, the HR group exhibited increased serum activities of SOD (20.26%), GSH-Px (9.03%), and CAT (133.01%), with a 24.87% decrease in MDA level. In brain tissue, SOD, GSH-Px, and CAT activities increased by 79.49%, 8.45%, and 60.23%, respectively, while MDA decreased by 8.29%. R. mucilaginosa JAASSRY also dose-dependently alleviated structural damage in the hippocampus and spleen and improved motor strength and learning-memory capacity. Furthermore, R. mucilaginosa JAASSRY increased the abundance of Lactobacillus and reduced Proteobacteria, Helicobacter, and Oscillospira, while enhancing antioxidant capacity by modulating nucleotide, lipid, and carbohydrate metabolism. Lactobacillus and Pediococcus were positively correlated with memory latency and CAT/SOD activities (p < 0.05), whereas Actinormyces and Dehalobacterium showed negative correlations. Notably, HR performed comparably or superiorly to β-carotene in improving cerebral oxidative stress and beneficial microbiota, suggesting its potential in neuroprotection and gut–brain axis regulation. In conclusion, R. mucilaginosa JAASSRY mitigates aging-related oxidative damage and behavioral deficits by modulating gut microbiota structure and function, demonstrating its promise as a β-carotene alternative in animal husbandry and functional foods.
{"title":"Rhodotorula mucilaginosa JAASSRY Alleviated Oxidative Damage in D-Galactose-Induced Aging Mice by Modulating the Gut Microbiota","authors":"Fenghao An, Yanchun Feng, Da Li, Mei Hua, Xiuquan Wang, Xifei Xu, Yuguang He, Xinyu Miao, Mubai Sun, Honghong Niu, Hongyan Xu, Jinghui Wang","doi":"10.3390/fermentation12010024","DOIUrl":"https://doi.org/10.3390/fermentation12010024","url":null,"abstract":"As one of the few carotenoid-producing microorganisms, Rhodotorula mucilaginosa remains underexplored for its antioxidant activity. This study investigated the effects of R. mucilaginosa JAASSRY on D-galactose-induced aging mice. The high-dose JAASSRY (HR) significantly increased body weight by 9.89% compared to the model group (AM), while reducing organ indices of the spleen, liver, kidneys, and brain (p < 0.01). Compared with the AM group, the HR group exhibited increased serum activities of SOD (20.26%), GSH-Px (9.03%), and CAT (133.01%), with a 24.87% decrease in MDA level. In brain tissue, SOD, GSH-Px, and CAT activities increased by 79.49%, 8.45%, and 60.23%, respectively, while MDA decreased by 8.29%. R. mucilaginosa JAASSRY also dose-dependently alleviated structural damage in the hippocampus and spleen and improved motor strength and learning-memory capacity. Furthermore, R. mucilaginosa JAASSRY increased the abundance of Lactobacillus and reduced Proteobacteria, Helicobacter, and Oscillospira, while enhancing antioxidant capacity by modulating nucleotide, lipid, and carbohydrate metabolism. Lactobacillus and Pediococcus were positively correlated with memory latency and CAT/SOD activities (p < 0.05), whereas Actinormyces and Dehalobacterium showed negative correlations. Notably, HR performed comparably or superiorly to β-carotene in improving cerebral oxidative stress and beneficial microbiota, suggesting its potential in neuroprotection and gut–brain axis regulation. In conclusion, R. mucilaginosa JAASSRY mitigates aging-related oxidative damage and behavioral deficits by modulating gut microbiota structure and function, demonstrating its promise as a β-carotene alternative in animal husbandry and functional foods.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"12 1","pages":"24-24"},"PeriodicalIF":0.0,"publicationDate":"2026-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/12/1/24/pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147381900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-27DOI: 10.3390/fermentation12010015
X. Zhang, Yi Liu, M. Zhang, Liyuan Chang, Yiqi QIN, Yaoxia Zhang
To optimize and model the fermentation process of Beauveria bassiana, adsorbed carrier solid-state fermentation (ACSSF) was used with rice husk as the inert support. The sample pretreating method was improved by combining homogenization and ultrasonic treatment after dry crushing; the large particles (100–1000 μm in size) were broken and the content of small particles (2–100 μm in size) increased, and the relative standard deviation of the biomass detection method was as low as 3.32% (intra-day) and 3.75% (inter-day). The most suitable carbon source—cassava starch—and the most suitable nitrogen source—corn steep liquor powder (CSLP)—were screened from multiple carbon and nitrogen sources. Through single-factor optimization and an artificial neural network combining genetic algorithm optimization, the optimal recipe including cassava starch 0.0314 g·cm−3, CSLP 0.004885 g·cm−3 and water 0.2630 g·cm−3 was obtained, and the highest biomass yield was verified as 0.1379 g·cm−3, which was 45.0% higher than the original recipe before the optimization (0.0951 g·cm−3). The modeling of microbial growth was based on the Logistic model and executed by nonlinear regression with the R2 value as high as 0.9525 and absolute value of the residues completely under 0.003 g·cm−3, which validated not only the feasibility of modeling the growth kinetics of B. bassiana using total biomass content, but also the reliability of the improved biomass pretreating and determination method.
{"title":"Adsorbed Carrier Solid-State Fermentation of Beauveria bassiana: Process Optimization and Growth Dynamics Modelization Based on an Improved Biomass Determination Method","authors":"X. Zhang, Yi Liu, M. Zhang, Liyuan Chang, Yiqi QIN, Yaoxia Zhang","doi":"10.3390/fermentation12010015","DOIUrl":"https://doi.org/10.3390/fermentation12010015","url":null,"abstract":"To optimize and model the fermentation process of Beauveria bassiana, adsorbed carrier solid-state fermentation (ACSSF) was used with rice husk as the inert support. The sample pretreating method was improved by combining homogenization and ultrasonic treatment after dry crushing; the large particles (100–1000 μm in size) were broken and the content of small particles (2–100 μm in size) increased, and the relative standard deviation of the biomass detection method was as low as 3.32% (intra-day) and 3.75% (inter-day). The most suitable carbon source—cassava starch—and the most suitable nitrogen source—corn steep liquor powder (CSLP)—were screened from multiple carbon and nitrogen sources. Through single-factor optimization and an artificial neural network combining genetic algorithm optimization, the optimal recipe including cassava starch 0.0314 g·cm−3, CSLP 0.004885 g·cm−3 and water 0.2630 g·cm−3 was obtained, and the highest biomass yield was verified as 0.1379 g·cm−3, which was 45.0% higher than the original recipe before the optimization (0.0951 g·cm−3). The modeling of microbial growth was based on the Logistic model and executed by nonlinear regression with the R2 value as high as 0.9525 and absolute value of the residues completely under 0.003 g·cm−3, which validated not only the feasibility of modeling the growth kinetics of B. bassiana using total biomass content, but also the reliability of the improved biomass pretreating and determination method.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"12 1","pages":"15-15"},"PeriodicalIF":0.0,"publicationDate":"2025-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/12/1/15/pdf?version=1766826609","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147330809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-28DOI: 10.3390/fermentation11120667
Yan Lü, Huayang Tang, Dexun Fan, Qingzhu Wang, Shuangyan Han
D-Allulose is a promising low-calorie rare sugar with significant health benefits. However, its industrial production is hindered by the low catalytic efficiency (≤33% conversion) and unfavorable equilibrium of the key enzyme, D-allulose 3-epimerase (DAE). To overcome this thermodynamic bottleneck, an in vitro synthetic enzymatic cascade based on a phosphorylation–dephosphorylation strategy was constructed. This engineered system comprises four synergistically operating enzymes: D-allulose-3-epimerase (DAE), L-rhamnulose kinase (RhaB), polyphosphate kinase (PPK), and acid phosphatase (AP). Through rational design and systematic optimization, the cascade achieved an exceptional 84.5% conversion yield from 50 mM D-fructose. Importantly, the system also maintained high conversion rates of 64.4% and 61.1% at high D-fructose loadings (50–100 g L−1). This performance, together with the integration of a low-cost PolyP6–PPK ATP regeneration module, underscores the potential industrial applicability of the proposed cascade strategy.
D-Allulose是一种很有前途的低热量稀有糖，对健康有很大的好处。但其催化效率低（转化率≤33%），关键酶D-allulose 3-epimerase （DAE）平衡不佳，阻碍了其工业化生产。为了克服这一热力学瓶颈，构建了基于磷酸化-去磷酸化策略的体外合成酶级联。该工程系统包括四种协同作用的酶：d -allulose-3- epimase (DAE)， l -鼠李糖激酶（RhaB），多磷酸激酶（PPK）和酸性磷酸酶（AP）。通过合理设计和系统优化，该级联从50 mM d -果糖中获得了84.5%的转化率。重要的是，在高d -果糖负荷（50-100 g L−1）下，该系统还保持了64.4%和61.1%的高转化率。这一性能，加上低成本的PolyP6-PPK ATP再生模块的集成，强调了所提出的级联策略的潜在工业适用性。
{"title":"An Engineered Multi-Enzyme Cascade with Low-Cost ATP Regeneration for Efficient D-Allulose Production from D-Fructose","authors":"Yan Lü, Huayang Tang, Dexun Fan, Qingzhu Wang, Shuangyan Han","doi":"10.3390/fermentation11120667","DOIUrl":"https://doi.org/10.3390/fermentation11120667","url":null,"abstract":"D-Allulose is a promising low-calorie rare sugar with significant health benefits. However, its industrial production is hindered by the low catalytic efficiency (≤33% conversion) and unfavorable equilibrium of the key enzyme, D-allulose 3-epimerase (DAE). To overcome this thermodynamic bottleneck, an in vitro synthetic enzymatic cascade based on a phosphorylation–dephosphorylation strategy was constructed. This engineered system comprises four synergistically operating enzymes: D-allulose-3-epimerase (DAE), L-rhamnulose kinase (RhaB), polyphosphate kinase (PPK), and acid phosphatase (AP). Through rational design and systematic optimization, the cascade achieved an exceptional 84.5% conversion yield from 50 mM D-fructose. Importantly, the system also maintained high conversion rates of 64.4% and 61.1% at high D-fructose loadings (50–100 g L−1). This performance, together with the integration of a low-cost PolyP6–PPK ATP regeneration module, underscores the potential industrial applicability of the proposed cascade strategy.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"11 12","pages":"667-667"},"PeriodicalIF":0.0,"publicationDate":"2025-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147333928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-14DOI: 10.3390/fermentation11110643
Caiyun Fang, Jiaqing Wang, Shuang Ma, Wenzhong Huang, X. Y. Liu, Max M. He, Fei He, Junfan Fu
The non-medicinal parts of Polygonatum sibiricum (P. sibiricum) and Gentiana scabra (G. scabra) are abundant but underutilized in Liaoning Province, China, creating an environmental burden. Solid-state fermentation (SSF) offers a strategy to enhance their bioactivity, yet triple microbial co-fermentation remains underexplored. This study applied a triple microbiota—featuring Aspergillus niger (A. niger), Bacillus subtilis (B. subtilis), and Saccharomyces cerevisiae (S. cerevisiae)—to ferment the stems and leaves of both plants. Antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) was assessed via the Kirby–Bauer test, while Liquid Chromatography–Tandem Mass Spectrometry (LC–MS/MS)-based non-targeted metabolomics identified differential metabolites and enriched pathways. Co-fermentation significantly increased the inhibition zones to 17.4 ± 0.8 mm for E. coli and 17.7 ± 0.3 mm for S. aureus, a 1.8-fold improvement over the unfermented controls (p < 0.001). Among the 2976 metabolites detected, 1236 were differentially expressed, with Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis highlighting activation of aminoacyl-tRNA biosynthesis, ABC transporter, and phenylalanine–tyrosine–tryptophan pathways. Differential abundance analysis indicated that the aminoacyl-tRNA pathway (DA score > 0.9) is critical for antimicrobial peptide synthesis. Phenylalanine derivatives, including 4-hydroxybenzaldehyde, which increased over 430-fold (Log2 FC = 8.78), contributed to membrane-disruptive antibacterial effects. Mechanistically, A. niger hydrolyzes cellulose to release precursors, B. subtilis synthesizes antimicrobial peptides, and S. cerevisiae enhances metabolite solubility and excretion, collectively boosting antibacterial activity by 80%, suggesting a potent synergistic interaction among the triple microbiota. This cascade mechanism provides a scalable approach for valorizing approximately 55 million tons of traditional Chinese medicine (TCM) waste annually.
{"title":"Valorization of the Non-Medicinal Parts of Polygonatum sibiricum and Gentiana scabra Bunge from Liaoning via Solid-State Co-Fermentation: Synergistic Antibacterial Enhancement","authors":"Caiyun Fang, Jiaqing Wang, Shuang Ma, Wenzhong Huang, X. Y. Liu, Max M. He, Fei He, Junfan Fu","doi":"10.3390/fermentation11110643","DOIUrl":"https://doi.org/10.3390/fermentation11110643","url":null,"abstract":"The non-medicinal parts of Polygonatum sibiricum (P. sibiricum) and Gentiana scabra (G. scabra) are abundant but underutilized in Liaoning Province, China, creating an environmental burden. Solid-state fermentation (SSF) offers a strategy to enhance their bioactivity, yet triple microbial co-fermentation remains underexplored. This study applied a triple microbiota—featuring Aspergillus niger (A. niger), Bacillus subtilis (B. subtilis), and Saccharomyces cerevisiae (S. cerevisiae)—to ferment the stems and leaves of both plants. Antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) was assessed via the Kirby–Bauer test, while Liquid Chromatography–Tandem Mass Spectrometry (LC–MS/MS)-based non-targeted metabolomics identified differential metabolites and enriched pathways. Co-fermentation significantly increased the inhibition zones to 17.4 ± 0.8 mm for E. coli and 17.7 ± 0.3 mm for S. aureus, a 1.8-fold improvement over the unfermented controls (p < 0.001). Among the 2976 metabolites detected, 1236 were differentially expressed, with Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis highlighting activation of aminoacyl-tRNA biosynthesis, ABC transporter, and phenylalanine–tyrosine–tryptophan pathways. Differential abundance analysis indicated that the aminoacyl-tRNA pathway (DA score > 0.9) is critical for antimicrobial peptide synthesis. Phenylalanine derivatives, including 4-hydroxybenzaldehyde, which increased over 430-fold (Log2 FC = 8.78), contributed to membrane-disruptive antibacterial effects. Mechanistically, A. niger hydrolyzes cellulose to release precursors, B. subtilis synthesizes antimicrobial peptides, and S. cerevisiae enhances metabolite solubility and excretion, collectively boosting antibacterial activity by 80%, suggesting a potent synergistic interaction among the triple microbiota. This cascade mechanism provides a scalable approach for valorizing approximately 55 million tons of traditional Chinese medicine (TCM) waste annually.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"11 11","pages":"643-643"},"PeriodicalIF":0.0,"publicationDate":"2025-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147333440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-04DOI: 10.3390/fermentation11110629
Xiaosi Lin, P. F. Wu, Y. S. Huang, Congjie Dai
Cycloheptylprodigiosin exhibits potent anticancer activity through a unique mechanism involving the induction of severe Golgi stress, a previously unrecognized cell death pathway. To increase the prodiginine production from the marine bacterium Spartinivicinus ruber MCCC 1K03745T, we investigated key fermentation parameters, including incubation time and initial medium pH. The culture medium composition was then sequentially optimized by single-factor experiments, a full factorial design, and an orthogonal design. Our results showed that the optimal incubation time was 30 h post inoculation, while initial pH had no effect on prodiginine production within the range of pH 6.0 to 8.0. By orthogonal design, the optimal medium was determined as follows: peptone 11 g/L, yeast extract 1 g/L, soybean oil 5 mL/L and MgCl2·6H2O 3 g/L in seawater. Verification experiments showed that prodiginine concentration under the optimized conditions reached 14.64 mg/L, representing 2.62 times the concentration obtained in basal Marine Broth 2216. These findings provide a basis for the cost-effective production of prodiginines from S. ruber MCCC 1K03745T for potential pharmaceutical applications.
{"title":"Optimization of Fermentation Parameters and Medium Composition for Producing Prodiginines from Marine Bacterium Spartinivicinus ruber MCCC 1K03745T","authors":"Xiaosi Lin, P. F. Wu, Y. S. Huang, Congjie Dai","doi":"10.3390/fermentation11110629","DOIUrl":"https://doi.org/10.3390/fermentation11110629","url":null,"abstract":"Cycloheptylprodigiosin exhibits potent anticancer activity through a unique mechanism involving the induction of severe Golgi stress, a previously unrecognized cell death pathway. To increase the prodiginine production from the marine bacterium Spartinivicinus ruber MCCC 1K03745T, we investigated key fermentation parameters, including incubation time and initial medium pH. The culture medium composition was then sequentially optimized by single-factor experiments, a full factorial design, and an orthogonal design. Our results showed that the optimal incubation time was 30 h post inoculation, while initial pH had no effect on prodiginine production within the range of pH 6.0 to 8.0. By orthogonal design, the optimal medium was determined as follows: peptone 11 g/L, yeast extract 1 g/L, soybean oil 5 mL/L and MgCl2·6H2O 3 g/L in seawater. Verification experiments showed that prodiginine concentration under the optimized conditions reached 14.64 mg/L, representing 2.62 times the concentration obtained in basal Marine Broth 2216. These findings provide a basis for the cost-effective production of prodiginines from S. ruber MCCC 1K03745T for potential pharmaceutical applications.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"11 11","pages":"629-629"},"PeriodicalIF":0.0,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/11/11/629/pdf?version=1762259333","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147332162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-21DOI: 10.3390/fermentation11100600
Suyang Li, Lhundrup Namgyal, Shiyi Chen, Yong Zhou, Afira Nayab, Qin Zhou, Dawa Dondup, Ling Sun
β-Glucan from Tibetan highland barley (THB) is an excellent edible gel polysaccharide due to its unique hypoglycemic and antioxidant activities. However, direct extraction of β-glucan from THB exhibits low yields with higher costs. Given that highland barley spent grain (BSG) is a byproduct of the brewing process and is frequently considered waste, the efficient extraction of its β-glucan could promote high-value repurposing of BSG. In this study, 2.74% β-glucan (BSG-B) was extracted from Rhizopus oryzae (R. oryzae)-fermented BSG, which is lower than those from THB (THB-B: 4.62%) yet enabled value-added utilization of BSG. The molecular weight of BSG-B was 5.24 × 106 Da, which significantly increased by 124.89% compared to that of THB-B. Fourier-transform infrared (FT-IR) spectroscopy showed similar absorption peaks in BSG-B and THB-B, except for structural modifications in the β-glucan pyranose ring induced by the fermentation of R. oryzae. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) indicated that BSG-B possesses a more compact structure and lower aggregation heights compared to THB-B. Moreover, BSG-B demonstrated superior antioxidant capacities to THB-B in NO/DPPH/ABTS/reducing power assays, and lower apparent viscosity and oil adsorption capacity, likely attributed to the fermentation of R. oryzae. This study establishes a foundation for extracting higher-molecular-weight antioxidant β-glucan from BSG.
{"title":"β-Glucan from Highland Barley Spent Grain: Yield, Molecular Weight, Physicochemical Properties, Antioxidant Capacity, and Gel Characteristics","authors":"Suyang Li, Lhundrup Namgyal, Shiyi Chen, Yong Zhou, Afira Nayab, Qin Zhou, Dawa Dondup, Ling Sun","doi":"10.3390/fermentation11100600","DOIUrl":"https://doi.org/10.3390/fermentation11100600","url":null,"abstract":"β-Glucan from Tibetan highland barley (THB) is an excellent edible gel polysaccharide due to its unique hypoglycemic and antioxidant activities. However, direct extraction of β-glucan from THB exhibits low yields with higher costs. Given that highland barley spent grain (BSG) is a byproduct of the brewing process and is frequently considered waste, the efficient extraction of its β-glucan could promote high-value repurposing of BSG. In this study, 2.74% β-glucan (BSG-B) was extracted from Rhizopus oryzae (R. oryzae)-fermented BSG, which is lower than those from THB (THB-B: 4.62%) yet enabled value-added utilization of BSG. The molecular weight of BSG-B was 5.24 × 106 Da, which significantly increased by 124.89% compared to that of THB-B. Fourier-transform infrared (FT-IR) spectroscopy showed similar absorption peaks in BSG-B and THB-B, except for structural modifications in the β-glucan pyranose ring induced by the fermentation of R. oryzae. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) indicated that BSG-B possesses a more compact structure and lower aggregation heights compared to THB-B. Moreover, BSG-B demonstrated superior antioxidant capacities to THB-B in NO/DPPH/ABTS/reducing power assays, and lower apparent viscosity and oil adsorption capacity, likely attributed to the fermentation of R. oryzae. This study establishes a foundation for extracting higher-molecular-weight antioxidant β-glucan from BSG.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"11 10","pages":"600-600"},"PeriodicalIF":0.0,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/11/10/600/pdf?version=1761047227","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147331326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-08DOI: 10.3390/fermentation11090525
H.-L. Xie, Bin Zhong, Qimin Zhang, Xi Hu, Xuesen Xia, Xie Hong, Zhenqiang Wu
Following new trends in green development, many studies have focused on the high-value utilization of fish resources through green biological processes. This study innovatively introduced a one-step process of mixed strain–enzyme synergy (MES) with which to prepare tilapia hydrolysates and explored the synergistic effects of strains and enzymes on both the protein hydrolysis process and its products’ characteristics via comparative experiments. Further, soybean was used as a model crop to verify the agronomic effects of the hydrolysates. The addition of exogenous papain increased hydrolysis by 31.94% compared to the fermentation-only group. Peptides and amino acids contents in the mixed strains were higher than those in the single fermentation process (p < 0.05), while 8.46 mg/L of indoleacetic acid was produced through fermentation. Hydrolysates promoted the growth of lateral roots in soybean seedlings (p < 0.05) via the use of a 2500-fold dilution of the biostimulant, increasing the root area and stem length and reducing the sugar content of soybean seedlings by 1.59-, 1.44- and 1.69-fold compared to those in Hoagland’s nutrient solution. These results lay a foundation for the biological preparation of biostimulants for hydroponic vegetables through the utilization of fish waste resources, aligning with green development goals.
{"title":"Transforming Tilapia into Indoleacetic Acid-Containing Biostimulants: Synergistic Effect of Enzymolysis and Multi-Strain Fermentation","authors":"H.-L. Xie, Bin Zhong, Qimin Zhang, Xi Hu, Xuesen Xia, Xie Hong, Zhenqiang Wu","doi":"10.3390/fermentation11090525","DOIUrl":"https://doi.org/10.3390/fermentation11090525","url":null,"abstract":"Following new trends in green development, many studies have focused on the high-value utilization of fish resources through green biological processes. This study innovatively introduced a one-step process of mixed strain–enzyme synergy (MES) with which to prepare tilapia hydrolysates and explored the synergistic effects of strains and enzymes on both the protein hydrolysis process and its products’ characteristics via comparative experiments. Further, soybean was used as a model crop to verify the agronomic effects of the hydrolysates. The addition of exogenous papain increased hydrolysis by 31.94% compared to the fermentation-only group. Peptides and amino acids contents in the mixed strains were higher than those in the single fermentation process (p < 0.05), while 8.46 mg/L of indoleacetic acid was produced through fermentation. Hydrolysates promoted the growth of lateral roots in soybean seedlings (p < 0.05) via the use of a 2500-fold dilution of the biostimulant, increasing the root area and stem length and reducing the sugar content of soybean seedlings by 1.59-, 1.44- and 1.69-fold compared to those in Hoagland’s nutrient solution. These results lay a foundation for the biological preparation of biostimulants for hydroponic vegetables through the utilization of fish waste resources, aligning with green development goals.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"11 9","pages":"525-525"},"PeriodicalIF":0.0,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/11/9/525/pdf?version=1757332225","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147333289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-28DOI: 10.3390/fermentation11090505
Di Chen, Yanping Xu, Han Li, Xuemin Zhu
Monascus pigments (MPs) are the most valuable secondary metabolites of Monascus. To improve the production of MPs is of great importance to food processing. Currently, studies using rice bran as the substrate to produce MPs are rare. In this study, rice bran with different carbon sources and cellulase hydrolysis conditions were explored in Monascus purpureus M9 in this study. Through single-factor experiments and Box–Behnken response surface optimization, we demonstrated that mannitol supplementation combined with cellulase treatment of substrate significantly enhanced the yields of MPs. The optimal conditions (4.00% mannitol, cellulase hydrolysis at 60 °C for 2 h) achieved a maximum color value of 3538 U/g. Furthermore, comparative evaluation under different culture conditions, including only rice bran (RB), cellulase hydrolysis of rice bran (Cel), rice bran supplemented with mannitol (Man), mannitol supplementation combined with cellulase pretreatment of substrate (Opti), and only rice (Rice), confirmed the effectiveness of the optimized treatment. The color value of the Opti group was 27.95 times more than that of the RB group and reached 80.96% of the counterpart of the Rice group. The Opti group also significantly enhanced the yields of two orange pigments (Monascorubrin and Rubropunctatin), induced more sexual spore formation, and exhibited the maximum biomass and colony diameter among different groups. The hyphae of the Man and Opti groups were full, intact, and tubular. The citrinin content in the Opti group was under the limit standard of China. The data provides a theoretical basis reference for improving the yields of MPs with RB as the substrate.
{"title":"Optimization of Monascus purpureus Culture Conditions in Rice Bran for Enhanced Monascus Pigment Biosynthesis","authors":"Di Chen, Yanping Xu, Han Li, Xuemin Zhu","doi":"10.3390/fermentation11090505","DOIUrl":"https://doi.org/10.3390/fermentation11090505","url":null,"abstract":"Monascus pigments (MPs) are the most valuable secondary metabolites of Monascus. To improve the production of MPs is of great importance to food processing. Currently, studies using rice bran as the substrate to produce MPs are rare. In this study, rice bran with different carbon sources and cellulase hydrolysis conditions were explored in Monascus purpureus M9 in this study. Through single-factor experiments and Box–Behnken response surface optimization, we demonstrated that mannitol supplementation combined with cellulase treatment of substrate significantly enhanced the yields of MPs. The optimal conditions (4.00% mannitol, cellulase hydrolysis at 60 °C for 2 h) achieved a maximum color value of 3538 U/g. Furthermore, comparative evaluation under different culture conditions, including only rice bran (RB), cellulase hydrolysis of rice bran (Cel), rice bran supplemented with mannitol (Man), mannitol supplementation combined with cellulase pretreatment of substrate (Opti), and only rice (Rice), confirmed the effectiveness of the optimized treatment. The color value of the Opti group was 27.95 times more than that of the RB group and reached 80.96% of the counterpart of the Rice group. The Opti group also significantly enhanced the yields of two orange pigments (Monascorubrin and Rubropunctatin), induced more sexual spore formation, and exhibited the maximum biomass and colony diameter among different groups. The hyphae of the Man and Opti groups were full, intact, and tubular. The citrinin content in the Opti group was under the limit standard of China. The data provides a theoretical basis reference for improving the yields of MPs with RB as the substrate.","PeriodicalId":507249,"journal":{"name":"Fermentation","volume":"11 9","pages":"505-505"},"PeriodicalIF":0.0,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.mdpi.com/2311-5637/11/9/505/pdf?version=1756374150","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147330898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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