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Fermentation
Volume 11
Issue 5
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Fermentation, Volume 11, Issue 5 (May 2025) – 9 articles

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17 pages, 4633 KiB  
Article
High-Level Extracellular Expression of Collagenase ColH in Bacillus subtilis for Adipose-Derived Cells Extraction
by Ling-Feng Xu, Dai Xue, Nuo Chen, Chang Su, Jin-Song Gong, Jian-Ying Qian, Zhen-Zhen Wang, Xu-Dong Ma, Nan Xie, Zheng-Hong Xu and Jin-Song Shi
Fermentation 2025, 11(5), 242; https://doi.org/10.3390/fermentation11050242 - 24 Apr 2025
Abstract
Collagenase has a wide range of applications in the medicine, cosmetic, and food industries. Inefficient expression of collagenase impedes its industrial production and commercial applications. In this study, a secretory expression system for collagenase ColH from Clostridium histolyticum was constructed in Bacillus subtilis [...] Read more.
Collagenase has a wide range of applications in the medicine, cosmetic, and food industries. Inefficient expression of collagenase impedes its industrial production and commercial applications. In this study, a secretory expression system for collagenase ColH from Clostridium histolyticum was constructed in Bacillus subtilis. Signal peptide optimization effectively solved the secretion problem of large collagenase with a molecular weight of about 116 kDa, doubling the extracellular enzyme activity. Then, promoter optimization further improved the enzyme activity to 264 U/mL. By the co-optimization of the nitrogen sources and carbon sources, and employing a fed-batch fermentation strategy, the enzyme activity could reach 669 U/mL, which is, currently, the highest level reported in the industry. The recombinant collagenase ColH was purified through a purification process suitable for industrial production with a specific activity of 565.25 U/mg. Based on the purified collagenase, cells were successfully prepared from adipose tissue, indicating its potential use in cell therapy. This study provides a promising candidate for the industrial production of collagenase and highlights its potential application to extract cells from tissues. Full article
(This article belongs to the Special Issue Applied Microorganisms and Industrial/Food Enzymes, 2nd Edition)
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17 pages, 306 KiB  
Review
Comprehensive Review of Strategies for Lactic Acid Bacteria Production and Metabolite Enhancement in Probiotic Cultures: Multifunctional Applications in Functional Foods
by Jiun Shen Loo, Siti Nur Hazwani Oslan, Nur Anis Safiah Mokshin, Rafidah Othman, Zarina Amin, Wipawee Dejtisakdi, Asep Awaludin Prihanto and Joo Shun Tan
Fermentation 2025, 11(5), 241; https://doi.org/10.3390/fermentation11050241 - 24 Apr 2025
Abstract
Lactic acid bacteria (LAB) play a crucial role in probiotics, functional foods, and sustainable biotechnologies due to their ability to produce bioactive metabolites such as short-chain fatty acids, bacteriocins, vitamins, and exopolysaccharides. These metabolites aid in gut health, pathogen inhibition, and enhanced productivity [...] Read more.
Lactic acid bacteria (LAB) play a crucial role in probiotics, functional foods, and sustainable biotechnologies due to their ability to produce bioactive metabolites such as short-chain fatty acids, bacteriocins, vitamins, and exopolysaccharides. These metabolites aid in gut health, pathogen inhibition, and enhanced productivity in the food, pharmaceutical, and aquaculture industries. However, the high production cost remains a major challenge, necessitating cost-effective media formulations and bioprocess optimization. This review explores strategies for maximizing LAB yields and functionality through the precision control of key cultivation parameters, including temperature, pH, and agitation speed, ensuring probiotic viability in compliance with regulatory standards (≥106 CFU/g or mL). Furthermore, advances in metabolic engineering, synthetic biology, and the utilization of agro-industrial by-products are driving cost-efficient and eco-friendly LAB production. By integrating scalable fermentation technologies with sustainable resource management, LAB have the potential to bridge the gap between food security, environmental sustainability, and biotechnological innovation. This review provides a comprehensive overview of recent advances in LAB cultivation and bioprocess optimization, ensuring high-quality probiotic production for diverse industrial applications. Full article
(This article belongs to the Section Fermentation for Food and Beverages)
16 pages, 2604 KiB  
Article
Exploring the Nutritional Profiling and Therapeutic Effect of Fermented Garlic on Alcohol-Induced Liver Injury in Animal Model
by Mavra Javed, Waqas Ahmed, Azmat Ullah and Imtiaz Rabbani
Fermentation 2025, 11(5), 240; https://doi.org/10.3390/fermentation11050240 - 24 Apr 2025
Abstract
Black garlic, a fermented product of fresh garlic, has shown promising potential as a culinary ingredient and a medicinal remedy. This study examined the microbiological makeup, nutritional profile, and health advantages of black garlic to better understand its health-endorsing properties. Thermus, Corynebacterium [...] Read more.
Black garlic, a fermented product of fresh garlic, has shown promising potential as a culinary ingredient and a medicinal remedy. This study examined the microbiological makeup, nutritional profile, and health advantages of black garlic to better understand its health-endorsing properties. Thermus, Corynebacterium, Streptococcus, and Brevundimonas were among the prominent taxa found when the microbial diversity in black garlic samples was investigated using Illumina MiSeq sequencing. This provided insight into the complex interactions between microorganisms during the fermentation process and clarified the distinctive qualities of black garlic. This study expanded its scope to include black garlic’s therapeutic potential, specifically in relation to liver function and hangovers caused by alcohol, in addition to its microbial complexity. Significant liver damage was revealed in alcohol-treated rats by serum biochemical indicators and histological stains; this damage was lessened by the administration of black garlic, particularly at higher dosages. Furthermore, black garlic showed hepatoprotective effects attributed to its high phenolic and flavonoid contents. These results offer a novel understanding of the medicinal qualities of black garlic as they lay out possibilities for the creation of functional drugs to treat alcohol-induced liver damage. Conclusively, black garlic’s diverse microbial composition also advances our knowledge of its nutritional makeup and health advantages. In summary, this research highlights the potential of black garlic as a flexible medical tool, having implications for both gastronomic and therapeutic uses. Full article
(This article belongs to the Special Issue Health and Bioactive Compounds of Fermented Foods and By-Products, 2.0 Version)
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27 pages, 4772 KiB  
Article
Optimized Amino Acid-Enhanced Medium for Efficient L-Asparaginase II Production in E. coli: From Shake Flask to Bioreactor
by Nicolás Lefin, Javiera Miranda, Iris Munhoz Costa, Alejandro Pedroso Reynaldo, Gisele Monteiro, Mauricio Zamorano, Adalberto Pessoa, Jr. and Jorge G. Farias
Fermentation 2025, 11(5), 239; https://doi.org/10.3390/fermentation11050239 - 23 Apr 2025
Abstract
L-asparaginase (L-ASNase) is a key enzyme in the treatment of leukemia and lymphoma, with high demand in cancer therapies. Advances in recombinant protein production have improved yields and reduced costs, enabling large-scale production. However, optimizing culture conditions remains crucial for maximizing production. This [...] Read more.
L-asparaginase (L-ASNase) is a key enzyme in the treatment of leukemia and lymphoma, with high demand in cancer therapies. Advances in recombinant protein production have improved yields and reduced costs, enabling large-scale production. However, optimizing culture conditions remains crucial for maximizing production. This study focused on optimizing the production of double mutant L-ASNase expressed in Escherichia coli BL21 (DE3) by supplementing media with amino acids. Five amino acids were evaluated at a shake flask scale using the design of experiments, with arginine and aspartate showing the most positive effects. Under optimized conditions (14.5 mM arginine, 12.7 mM aspartate, and 0 mM cysteine), the activity model reached 12,513 U L−1, experimentally validated at 10,089 U L−1. The maximum specific cell growth rate was µx,max = 0.74 h−1, with substrate–biomass conversion factor Yx/s = 1.16 g/g and cell–product conversion factor Yp/x = 13,891 U/gcell. Amino acid supplementation resulted in a ten-fold increase in L-ASNase activity. Finally, at the bioreactor scale, adding amino acids and the inducer at the end of the exponential phase increased activity by 135% compared to conventional MD, demonstrating its potential for industrial-scale production. Full article
(This article belongs to the Special Issue Research on Microbial Protein Synthesis: 2nd Edition)
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17 pages, 2382 KiB  
Article
Effects of Co-Fermentation with Lactic Acid Bacteria and Yeast on Gliadin Degradation in Whole-Wheat Sourdough
by Daiva Zadeike, Kamile Cipkute and Dalia Cizeikiene
Fermentation 2025, 11(5), 238; https://doi.org/10.3390/fermentation11050238 - 23 Apr 2025
Abstract
This study investigates the potential of utilising the proteolytic activity of two different strains, Levilactobacillus brevis FST140 and Pediococcus pentosaceus FST22, to assess their impact on wheat gluten proteins. A high-power ultrasound (US) treatment (850 kHz; 500 W/cm2; 35 °C) was [...] Read more.
This study investigates the potential of utilising the proteolytic activity of two different strains, Levilactobacillus brevis FST140 and Pediococcus pentosaceus FST22, to assess their impact on wheat gluten proteins. A high-power ultrasound (US) treatment (850 kHz; 500 W/cm2; 35 °C) was used to activate the proteolytic system of LAB to promote gliadin-like protein degradation in wheat wholemeal-based sourdough. The proteolytic activity of L. brevis and P. pentosaceus increased two-fold with 10 and 20 min US stimulation, respectively, compared to fermentation without ultrasonication. Regarding the impact of proteolysis and sonication on gliadin proteins, fermentation with both strains reduced gliadin content in commercial gluten by an average of 77.4% compared to the untreated sample, and additional US treatment further enhanced gliadin degradation efficiency to an average of 83.5%. The combined application of US and lactic acid fermentation initiated a seven-fold decrease in wheat wholemeal flour (WF) gliadin levels compared to the untreated sample (47.2 mg/g). Furthermore, the synergistic application of US, LAB, and yeast fermentation allowed us to reduce gliadin content up to 1.6 mg/g, as well as to reduce gluten content in the sourdough up to 3 mg/g. Despite complete hydrolysis of the gliadin fraction under the combined effects of US and fermentation, glutenins were less affected by the applied treatments in all cases. The technology presented in this study offers a promising approach for producing gluten-free or low-gluten fermented products in the bread-making industry. Full article
(This article belongs to the Special Issue Bioactive Compounds in Grain Fermentation: 2nd Edition)
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18 pages, 3307 KiB  
Article
Olive Pruning: Waste or Growth Media? Expanding the Metabolic Potential of Phyllospheric Rhodococcus sp. 24CO
by Natalia E. Sandoval, Margarita Gomila, Nadia S. Arias, Héctor M. Alvarez and Mariana P. Lanfranconi
Fermentation 2025, 11(5), 237; https://doi.org/10.3390/fermentation11050237 - 23 Apr 2025
Abstract
Rhodococcus sp. 24CO, isolated from the olive phyllosphere, can accumulate significant amounts of neutral lipids, making it a promising candidate for biomass production from olive pruning waste. The strain efficiently converts this residue to neutral lipids, achieving a yield of over 20% of [...] Read more.
Rhodococcus sp. 24CO, isolated from the olive phyllosphere, can accumulate significant amounts of neutral lipids, making it a promising candidate for biomass production from olive pruning waste. The strain efficiently converts this residue to neutral lipids, achieving a yield of over 20% of the cellular dry weight (CDW). This indicates that olive leaves, a by-product of the olive oil industry, could become a valuable resource for both the economy and the environment. Genome analysis revealed various metabolic pathways for converting carbon sources to neutral lipids, while phenotypic studies showed that the strain is selective about its carbon sources, thriving on specific monosaccharides and polyols found in olive leaves. Notably, fructose and mannitol were rapidly metabolized, leading to a content of stored triacylglycerides of up to 47% and 28% of the CDW, respectively. The strain also exhibited oleagenicity under high nitrogen availability when grown on mannitol. Finally, potential oleagenicity determinants were explored through an omics comparison. Full article
(This article belongs to the Special Issue Producing Lipids and Lipid Derivatives by Fermentation Using Agro-Industrial By-Products as Substrates)
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21 pages, 1128 KiB  
Review
Applications of Yeasts in Heavy Metal Remediation
by Qi Shao, Shihui Yan, Xin Sun, Hongming Chen, Yixiao Lu, Siqi Li, Yunjie Huang, Shimei Wang, Min Zhang and Zhen Li
Fermentation 2025, 11(5), 236; https://doi.org/10.3390/fermentation11050236 - 23 Apr 2025
Abstract
Yeasts have been extensively recognized as a type of model microorganism due to their facile cultivation, short growth cycle, and genetic stability. Different yeast strains, such as Saccharomyces cerevisiae and Rhodotorula mucilaginosa, have exhibited notable sorption capacities for heavy metals and metalloids. [...] Read more.
Yeasts have been extensively recognized as a type of model microorganism due to their facile cultivation, short growth cycle, and genetic stability. Different yeast strains, such as Saccharomyces cerevisiae and Rhodotorula mucilaginosa, have exhibited notable sorption capacities for heavy metals and metalloids. Yeast employs diverse pathways for detoxifying heavy metals via its cell walls, intracellular organelles, and extracellular polymeric substances (EPSs). The cell wall has many functional groups to adsorb metals, decreasing their concentrations in the environment. In intracellular regions, some proteins are capable of transporting metals into biological metabolic processes for detoxification. In extracellular regions, electrostatic as well as complexation mechanisms between protein in EPSs and heavy metals is well accepted. Meanwhile, mannose and glucose within EPSs are target sugars for complexation with metals. Many yeasts can hence work as excellent biomaterials for the bioremediation of metal pollution. Meanwhile, they can be combined with other materials to enhance remediation efficiency. This study reviews underlying mechanisms and cases of yeast-mediated metal detoxification, alongside highlighting yeasts’ industrial applications as bioremediation materials. Full article
(This article belongs to the Special Issue 10th Anniversary of Fermentation: Feature Papers in Section “Microbial Metabolism, Physiology & Genetics”)
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18 pages, 7962 KiB  
Article
Effects of Soybean Meal Fermented by Lactobacillus plantarum NX69 on Growth Performance and Intestinal Health of Nursery Pigs
by Mengshi Zhao, Fengqiang Lin, Song Peng, Yaxiong Ma, Huini Wu and Zhaolong Li
Fermentation 2025, 11(5), 235; https://doi.org/10.3390/fermentation11050235 - 22 Apr 2025
Abstract
The Intestinal system of nursery pigs is not fully matured and is easily disturbed by dietary composition and nutritional content. Probiotic-fermented feed has emerged as a beneficial dietary form for nursery pigs and is widely utilized in the livestock and poultry industries. However, [...] Read more.
The Intestinal system of nursery pigs is not fully matured and is easily disturbed by dietary composition and nutritional content. Probiotic-fermented feed has emerged as a beneficial dietary form for nursery pigs and is widely utilized in the livestock and poultry industries. However, there are limited reports regarding the effects of Lactobacillus plantarum-fermented feed on intestinal health and growth performance in nursery pigs. In this study, we investigated the effect of Lactobacillus plantarum NX69-fermented soybean by multi-omics approaches on intestinal health and growth performance in nursery pigs. The results demonstrated that NX69-fermented soybean meal increased small intestinal villus height, the villus height-to-crypt depth ratio (V/C), and the number of goblet cells per unit length. Additionally, it enhanced the mRNA of intestinal mucosal barrier factors ZO-1, Occludin, and Claudin in nursery pigs. Further research revealed that NX69-fermented soybean meal increased the diversity of the intestinal microbiota structure, elevated the abundance of core microbiota such as Alloprevotella, Prevotellaceae, and Megasphaera in the cecum, and increased the abundance of genera such as Megasphaera, Faecalibacterium, and Ruminococcus, which are known to produce short-chain fatty acids (SCFAs) in the cecum. Correlation analysis indicated that the core microbiota were positively correlated with intestinal physical barriers, including villus length and the V/C ratio, as well as with the mRNA level of intestinal mucosal barrier factors ZO-1, Occludin, and Claudin. Furthermore, they were positively correlated with differential metabolites such as Ginkgetin, Formiminoglutamic acid, Naringenin, and Hydroxyisocaproic acid. These findings suggest that NX69-fermented soybean meal can enhance the intestinal mucosal barrier in nursery pigs by increasing the abundance of core microbiota that produce SCFAs and then promoting intestinal health and improving growth performance, indicating promising application prospects. Full article
(This article belongs to the Special Issue 10th Anniversary of Fermentation: Feature Papers in Section "Probiotic Strains and Fermentation")
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50 pages, 1219 KiB  
Review
Bioactive Properties of Fermented Beverages: Wine and Beer
by Vanesa Postigo, Margarita García, Julia Crespo, Laura Canonico, Francesca Comitini and Maurizio Ciani
Fermentation 2025, 11(5), 234; https://doi.org/10.3390/fermentation11050234 - 22 Apr 2025
Abstract
In recent years, consumer demand has been increasingly oriented to fermented foods and/or beverages with functional properties. The functional beverage industry focused on producing a product that combines a peculiar aromatic taste with healthy properties. Today’s consumers are trying to reduce alcohol, gluten, [...] Read more.
In recent years, consumer demand has been increasingly oriented to fermented foods and/or beverages with functional properties. The functional beverage industry focused on producing a product that combines a peculiar aromatic taste with healthy properties. Today’s consumers are trying to reduce alcohol, gluten, sugar, and carbohydrates in beer and wine without reducing their native taste. Wine and beer are among the world’s most consumed beverages, and several studies confirm that fermented beverages could be associated with beneficial properties for human health. All beneficial properties derive both from the fermentation process and also from the characteristics of the raw materials used in the two beverages. This review was conducted to highlight the importance of the fermentative microorganisms in wine and beer and their relationship with functional foods, underlining their involvement in human health. Full article
(This article belongs to the Special Issue Biotechnological and Functional/Probiotic Characteristics of Non-Conventional Yeasts in Fermented Beverages)
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