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23 pages, 4580 KB  
Article
Bacillus velezensis 7-A as a Biocontrol Agent Against Fusarium verticillioides, the Causal Agent of Rice Sheath Rot Disease
by Boyu Liu, Qunying Qin, Jianchao Hu, Jiayi Wang, Juan Gan, Ye Zhuang, Zhengxiang Sun and Yi Zhou
Microorganisms 2025, 13(11), 2511; https://doi.org/10.3390/microorganisms13112511 (registering DOI) - 31 Oct 2025
Abstract
Rice sheath rot has progressively developed into a growing threat to global rice production, particularly in intensively managed systems conducive to disease development. Therefore, accurate identification of the causal pathogen and the development of sustainable management strategies represent urgent scientific requirements. In this [...] Read more.
Rice sheath rot has progressively developed into a growing threat to global rice production, particularly in intensively managed systems conducive to disease development. Therefore, accurate identification of the causal pathogen and the development of sustainable management strategies represent urgent scientific requirements. In this study, we isolated the causal organism of rice sheath rot from infected rice tissues and identified it as Fusarium verticillioides based on multi-locus sequence analysis. Eight endophytic bacterial strains were recovered from healthy rice root systems. Among the isolates, Bacillus velezensis isolate 7-A exhibited the strongest antifungal activity against F. verticillioides. This isolate demonstrated broad-spectrum antifungal activity, with inhibition rates ranging from 54.8% to 71.8%. Phylogenetic analysis based on 16S rRNA and gyrB gene sequences identified it as B. velezensis. Further characterization revealed that B. velezensis 7-A is capable of secreting proteases and synthesizing siderophores. The filtered liquid from sterile fermentation markedly inhibited the growth of mycelium in F. verticillioides and induced marked morphological abnormalities. Liquid LC-MS analysis identified multiple antifungal active substances, including camphor, ginkgolides B, salicin, cinnamic acid, hydroxygenkwanin, stearamide, β-carotene, and others. A pot experiment demonstrated that the fermentation broth of B. velezensis 7-A effectively suppressed the occurrence of rice sheath rot, achieving a relative control efficacy of 61.3%, which is comparable to that of a 10% carbendazim water-dispersible granule (WDG). Additionally, isolate 7-A enhances plant disease resistance by activating the activities of key defense enzymes. These findings provide preliminary insights into its potential application in integrated and sustainable disease management programs. Full article
(This article belongs to the Special Issue Beneficial Microorganisms for Sustainable Agriculture)
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18 pages, 998 KB  
Article
Production of Bio-Improved Butter with Lactic Acid Bacteria Isolated from Traditional Cheese Matrix and Eye Fluid
by Gokce Keser and Tulay Ozcan
Fermentation 2025, 11(11), 620; https://doi.org/10.3390/fermentation11110620 (registering DOI) - 31 Oct 2025
Abstract
This study aimed to investigate the effects of Levilactobacillus brevis, Lacticaseibacillus paracasei, and Lacticaseibacillus rhamnosus strains isolated from Mihalic cheese, also known as “weeping cheese”, on fermentation kinetics, microbial viability, and textural and aromatic properties of the butter matrix. The effects [...] Read more.
This study aimed to investigate the effects of Levilactobacillus brevis, Lacticaseibacillus paracasei, and Lacticaseibacillus rhamnosus strains isolated from Mihalic cheese, also known as “weeping cheese”, on fermentation kinetics, microbial viability, and textural and aromatic properties of the butter matrix. The effects of the isolates were determined on acidification kinetics (Vmax, Tvmax, pHvmax), viability proportion index (VPI), textural parameters (firmness, work of shear, stickiness, work of adhesion), and volatile aroma compounds (GC-MS) formation. This study found that the BLR sample containing Lacticaseibacillus rhamnosus maintained its limited viability under acidic stress conditions despite its high fermentation rate and low pHvmax values. The BLP sample containing Lacticaseibacillus paracasei exhibited high viability due to its low acidification rate and limited pH change. Determining the chemical classes to which the aroma compounds in the BLP sample belonged revealed a composition rich in fatty acids. The BLB sample containing Levilactobacillus brevis produced a high ΔpH value and an aroma profile rich in aldehyde compounds. Examination of the macro-structural properties of the butter samples revealed that the sample containing Lacticaseibacillus rhamnosus, similar to the control sample (BMC), was more compact and rigid during storage. In contrast, samples containing Lacticaseibacillus paracasei and Levilactobacillus brevis had a softer/spreadable texture. These findings demonstrate the potential of lactic acid bacteria isolates from the traditional Mihalic cheese microbiota as biological catalysts for the development/improvement of texture, aroma, and sensory quality in high-fat dairy products and for the industrial production of products modified to meet consumer preferences. Full article
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20 pages, 2802 KB  
Article
Development of a Mixed-Strain Fermentation Process for Sour Pomegranate: An Analysis of Metabolites and Flavor Compounds
by Yuting Yang, Ailikemu Mulati, Xinmeng Huang, Yuanpeng Li, Dilireba Shataer, Haipeng Liu and Jiayi Wang
Foods 2025, 14(21), 3733; https://doi.org/10.3390/foods14213733 - 30 Oct 2025
Abstract
Sour pomegranate, a distinctive product of Xinjiang, China, is characterized by its sour and astringent taste, which contributes to a low processing rate. This study utilized single-factor experiments to screen three strains: Lactobacillus fermentum, Lactobacillus plantarum, and Lactobacillus acidophilus. Through [...] Read more.
Sour pomegranate, a distinctive product of Xinjiang, China, is characterized by its sour and astringent taste, which contributes to a low processing rate. This study utilized single-factor experiments to screen three strains: Lactobacillus fermentum, Lactobacillus plantarum, and Lactobacillus acidophilus. Through uniform design experiments, the mixed-strain ratio of L. fermentum:L. plantarum:L. acidophilus = 45%:31%:28% was determined. In addition, the amount of mixed inoculum was 2%, and the fermentation time was 11 h. Additionally, a mixed inoculation amount of 2% and a fermentation duration of 11 h were established. Utilizing electronic nose, electronic tongue, gas chromatography–ion mobility spectrometry, and non-targeted metabolomics, comparative analyses were conducted on the flavors and metabolites pre- and post-fermentation. The findings indicated that post-fermentation, umami increased by 32%, richness was enhanced by 6%, and the positive aftertaste was significantly extended. Mixed-strain fermentation facilitated the enrichment of alkanes, alcohols, aldehydes, and terpene volatile compounds; notably, the content of hexanal (fresh fruity aroma) and limonene (citrus aroma) increased by 1.95 times and 1.45 times, respectively, thereby augmenting the complexity of the aroma. Furthermore, mixed-strain fermentation significantly upregulated terpenes, amino acids and their derivatives, steroids and their derivatives, and alkaloid metabolites. These results offer potential technical support for the high-value utilization of agricultural products. Full article
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18 pages, 1577 KB  
Article
Biopreservation of Hericium erinaceus By-Products via Lactic Acid Fermentation: Effects on Functional and Technological Properties
by Mafalda Silva, Manuela Vida, Ana Cristina Ramos, Luísa Cristina Roseiro, Nuno Alvarenga, Sandra Gomes, Fernando C. Lidon, Fernando H. Reboredo and Elsa M. Gonçalves
Foods 2025, 14(21), 3721; https://doi.org/10.3390/foods14213721 - 30 Oct 2025
Abstract
Lactic acid fermentation is an effective strategy for food preservation and functional enhancement. This study evaluated the fermentation of Hericium erinaceus by-products using Lactoplantibacillus plantarum and Lactocaseibacillus rhamnosus, assessing microbial stability, physicochemical parameters, phenolic content, antioxidant activity, rheology, and biogenic amine formation [...] Read more.
Lactic acid fermentation is an effective strategy for food preservation and functional enhancement. This study evaluated the fermentation of Hericium erinaceus by-products using Lactoplantibacillus plantarum and Lactocaseibacillus rhamnosus, assessing microbial stability, physicochemical parameters, phenolic content, antioxidant activity, rheology, and biogenic amine formation over 240 h. Lp. plantarum promoted rapid acidification, reducing the pH from 6.0 to 4.65 within 72 h, while Ls. rhamnosus reached its lowest value of 3.8 at 144 h. Both strains effectively inhibited spoilage organisms: Lp. plantarum suppressed yeasts, molds, and Pseudomonas spp. by 144 h, whereas the control reached >6.0 log CFU/g of Pseudomonas at 240 h. Fermentation altered the texture, with the storage modulus (G′) decreasing from ~17 kPa to <3 kPa. Functional enrichment was also observed, with total phenolic content increasing from 32 to 48 mg GAE/100 g and antioxidant activity (DPPH) reaching 2562 µmol TE/100 g compared with 1954 µmol TE/100 g in the control. Importantly, cadaverine accumulated to 70.3 mg/kg in the control but remained below 15 mg/kg in inoculated samples, while spermidine was consistently higher in Lp. plantarum-treated mushrooms (~45 mg/kg). These results demonstrate that lactic acid fermentation can transform perishable H. erinaceus by-products into safe, stable, and bioactive ingredients, supporting their application in functional foods, nutraceuticals, and clean-label products while contributing to circular bioeconomy goals. Full article
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16 pages, 641 KB  
Article
A Strategy for Sustainable Production of Isoflavones from Black Soybean Okara via Solid-State Co-Fermentation
by Yi-Chung Lai, Bang-Yuan Chen, Jung-Feng Hsieh, Chien-Cheng Yeh, Cheng Huang, Meng-I Kuo and Chun-Ping Lu
Fermentation 2025, 11(11), 619; https://doi.org/10.3390/fermentation11110619 - 29 Oct 2025
Viewed by 346
Abstract
Okara, the soybean residue generated during soymilk and tofu production, is nutrient-rich but underutilized due to its high moisture content and perishability. This study established a sustainable solid-state co-fermentation strategy using Rhizopus oligosporus (BCRC 31631) and Yarrowia lipolytica (BCRC 21252) to enhance the [...] Read more.
Okara, the soybean residue generated during soymilk and tofu production, is nutrient-rich but underutilized due to its high moisture content and perishability. This study established a sustainable solid-state co-fermentation strategy using Rhizopus oligosporus (BCRC 31631) and Yarrowia lipolytica (BCRC 21252) to enhance the bioactive value of black soybean okara from two Taiwanese cultivars—Tainan No. 3 (TN.3) and Tainan No. 5 (TN.5). Co-fermentation markedly enhanced β-glucosidase activity, reaching 0.75 U/g DW at 30 °C after 48 h in TN.3 and 0.68 U/g DW after 24 h in TN.5, approximately 3.5-fold higher than single-strain fermentation. Near-complete (97–100%) hydrolysis of daidzin, glycitin, and genistin occurred within 24–48 h, producing 672.9 µg/g DM of total aglycone-type isoflavones—an ~11-fold increase compared with unfermented okara (61.5 µg/g DM), where most isoflavones (~740 µg/g DM) remained glycosylated. Varietal structure affected conversion efficiency: TN.3 exhibited stronger enzymatic responsiveness and higher aglycone yield, whereas TN.5 showed faster but less extensive transformation. The process also revealed a bioactive–pigment trade-off, as enhanced isoflavone activation coincided with anthocyanin degradation. These findings demonstrate that ambient-temperature solid-state co-fermentation effectively boosts β-glucosidase activity and isoflavone bioconversion, transforming low-value okara into a high-value functional ingredient consistent with circular bioeconomy and sustainable food system goals. Full article
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21 pages, 862 KB  
Article
Polyphasic Characterisation of Microbiota Associated with Sant’Agostino Table Olives Flavoured with Foeniculum vulgare
by Antonio Alfonzo, Raimondo Gaglio, Davide Alongi, Elena Franciosi, Giulio Perricone, Giuliana Garofalo, Rosario Prestianni, Vincenzo Naselli, Antonino Pirrone, Nicola Francesca, Giancarlo Moschetti and Luca Settanni
Foods 2025, 14(21), 3689; https://doi.org/10.3390/foods14213689 - 29 Oct 2025
Viewed by 249
Abstract
Sant’Agostino green table olives, traditionally processed in Apulia and flavoured with Foeniculum vulgare, represent a niche product whose microbial ecology remains largely unexplored. This study aimed to characterise the microbiota of the final product (both brine and fruit) after six months of [...] Read more.
Sant’Agostino green table olives, traditionally processed in Apulia and flavoured with Foeniculum vulgare, represent a niche product whose microbial ecology remains largely unexplored. This study aimed to characterise the microbiota of the final product (both brine and fruit) after six months of storage with wild fennel. Four production batches were analysed using a combined culture-dependent and culture-independent approach. Microbiological counts revealed variable levels of aerobic mesophilic microorganisms, yeasts, lactic acid bacteria (LAB), and staphylococci, with yeasts and LAB being predominant. Ten LAB strains were identified, including Enterococcus faecium, Leuconostoc mesenteroides subsp. jonggajibkimchii, Leuconostoc mesenteroides subsp. cremoris, Leuconostoc pseudomesenteroides, Lactiplantibacillus plantarum, and Lactiplantibacillus pentosus. Yeast isolates belonged to Candida tropicalis, Torulaspora delbrueckii, and Saccharomyces cerevisiae. Amplicon sequencing (MiSeq Illumina) revealed distinct bacterial profiles between fruit and brine samples, with taxa from Actinobacteria, Bacteroidetes, Enterococcus, Lactobacillus, Leuconostoc, Alphaproteobacteria, Enterobacteriaceae, and other Gammaproteobacteria. Enterococcus and Leuconostoc were consistently detected, while Lactobacillus sensu lato appeared only in one fruit and one brine sample. These findings provide new insights into the microbial diversity of Sant’Agostino olives and contribute to the understanding of their fermentation ecology and potential for quality and safety enhancement. Full article
(This article belongs to the Special Issue Changes in Microbial Community Structure of Fermented Food)
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22 pages, 962 KB  
Article
Synergistic Evaluation of Lactobacilli on Probiotic Viability and Antioxidant Properties in Almond and Cow Milk
by Ashwag Jaman Al Zahrani, Amal Bakr Shori and Khadijah A. Altammar
Fermentation 2025, 11(11), 616; https://doi.org/10.3390/fermentation11110616 - 29 Oct 2025
Viewed by 260
Abstract
Lactobacilli are important probiotic groups recognized for their numerous health-promoting properties. This study investigated how four probiotic strains, Lacticaseibacillus rhamnosus (Lr), Lactobacillus acidophilus (La), Lactiplantibacillus plantarum (Lp), or Lacticaseibacillus casei (Lc), affected post-acidification, viable cell counts (VCCs), total phenolic and flavonoid contents (TPCs [...] Read more.
Lactobacilli are important probiotic groups recognized for their numerous health-promoting properties. This study investigated how four probiotic strains, Lacticaseibacillus rhamnosus (Lr), Lactobacillus acidophilus (La), Lactiplantibacillus plantarum (Lp), or Lacticaseibacillus casei (Lc), affected post-acidification, viable cell counts (VCCs), total phenolic and flavonoid contents (TPCs and TFCs, respectively), and antioxidant activity of fermented almond milk (FAM) and its combination with cow’s milk (CM) at different concentrations (75:25, 50:50, and 25:75) during 1, 7, 14, and 21 days of storage. All FAM and its mixture with CM showed significantly greater (p < 0.05) post-acidification than their respective controls throughout storage. Viable cell counts in all samples ranged from 5.9 to 6.8 log cfu/mL, which were higher than those of the controls (3–4 log cfu/mL; p < 0.05). Total phenolic contents in FAM/CM (75:25 and 50:50 and 25:75)-Lc increased more than twofold (95.82 ± 0.003 and 105.71 ± 0.008 and 101.02 ± 0.071 μg GAE/mL; p < 0.05) compared to the controls (19–40 μg GAE/mL) by the end of the third week. Lbs. rhamnosus enhanced (p < 0.05) TFCs in FAM/CM (25:75) after the first day of storage. All lactobacilli strains improved the antioxidant activity in all treated samples during storage. In conclusion, the combination of fermented almond milk with cow’s milk may serve as an excellent carrier for Lbs. rhamnosus, Lab. acidophilus, Lpb. plantarum, and Lbs. casei, which exhibit antioxidant activity. Full article
(This article belongs to the Section Probiotic Strains and Fermentation)
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18 pages, 2650 KB  
Article
Lychee Fermented by Mixed Probiotic Strains Alleviates D-Galactose-Induced Skeletal Muscle and Intestinal Aging in Mice
by Huixian Han, Jin Tao, Xiaoyue Bai, Yizhi Jing, Zhengyuan Zhai, Junjie Luo, Wanxiang Zhang, Dan Gan and Yanling Hao
Foods 2025, 14(21), 3684; https://doi.org/10.3390/foods14213684 - 29 Oct 2025
Viewed by 158
Abstract
Aging-associated skeletal muscle and intestinal dysfunction is largely driven by chronic inflammation, oxidative stress, and microbiota imbalance. This study investigated the protective effects of a lychee fermentate (LF) in a D-galactose-induced aging mouse model. LF was prepared using a mixed microbial fermentation approach [...] Read more.
Aging-associated skeletal muscle and intestinal dysfunction is largely driven by chronic inflammation, oxidative stress, and microbiota imbalance. This study investigated the protective effects of a lychee fermentate (LF) in a D-galactose-induced aging mouse model. LF was prepared using a mixed microbial fermentation approach with Lactiplantibacillus plantarum, Lacticaseibacillus casei, Saccharomyces cerevisiae, and Acetobacter pasteurianus SP021. LF administration significantly improved muscle strength and endurance and restored muscle fiber morphology. Meanwhile, LF alleviated colonic transit impairment and downregulated senescence markers p16 and p21. H&E and AB-PAS staining showed that intervention with LF ameliorated the colonic tissue damage, preserved goblet cell populations and promoted MUC2-mediated mucus secretion, which was further confirmed by the upregulation of intestinal barrier-related proteins MUC2, ZO-1, Claudin-1, and Occludin through immunofluorescence analysis. In addition, LF reduced colonic inflammation by suppressing IL-1β, IL-6, TNF-α, CXCL1, and MCP1 expression, and mitigated oxidative stress by lowering malondialdehyde levels to 24.65 ± 3.84 nmol/mL while enhancing glutathione peroxidase and superoxide dismutase activities. Moreover, the LF restored intestinal health by modulating microbiota homeostasis, such as adjusting the Firmicutes/Bacteroidetes ratio and increasing the abundance of beneficial bacteria like Clostridia_UCG-014 and Alistipes. Metabolomic profiling indicated that the enhanced bioactivity of the LF was primarily attributed to the enrichment of phenolic acids, flavonoids and their derivatives postfermentation, including ethyl caffeate, gallic acid, kaempferol and isorhamnetin. In summary, these findings provided new insights into the potential application of LF as a functional food for mitigating skeletal muscle and intestinal aging. Full article
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19 pages, 2453 KB  
Article
Safety Assessment and the Potential of a Postbiotic Powder Formulated from a Three-Strain Fermentation of Lactobacillus salivarius AP-32, Lactobacillus paracasei ET-66, and Lactobacillus plantarum LPL28
by Ching-Min Li, Yen-Yu Huang, Chi-Huei Lin, Jia-Hung Lin, Yi-Wei Kuo, Ko-Chiang Hsia, Shin-Yu Tsai, Yu-Fen Huang, Ching-Wei Chen and Hsieh-Hsun Ho
Fermentation 2025, 11(11), 615; https://doi.org/10.3390/fermentation11110615 - 28 Oct 2025
Viewed by 166
Abstract
This study evaluated the safety, antimicrobial activity, and upper gastrointestinal gastroprotection of a postbiotic powder derived from Lactobacillus salivarius AP-32, Lactobacillus paracasei ET-66, and Lactobacillus plantarum LPL28. Safety assessments were performed in rodent models through acute and subchronic oral toxicity tests, genotoxicity assays, [...] Read more.
This study evaluated the safety, antimicrobial activity, and upper gastrointestinal gastroprotection of a postbiotic powder derived from Lactobacillus salivarius AP-32, Lactobacillus paracasei ET-66, and Lactobacillus plantarum LPL28. Safety assessments were performed in rodent models through acute and subchronic oral toxicity tests, genotoxicity assays, and biogenic amine analysis. No signs of toxicity were observed in either the acute (20 g/kg body weight, BW) or subchronic (3 g/kg BW) toxicity tests. Genotoxicity evaluations indicated no mutagenic activity in the Ames test (≤5000 µg/plate) and no chromosomal or micronuclear abnormalities in the spermatocyte or the peripheral blood assays (≤10 g/kg BW). Biogenic amines were undetectable in the postbiotic powder, further reinforcing its safety. The postbiotic powder showed significant direct antimicrobial activity. Additionally, it enhanced the inhibitory effects of probiotics against key upper gastrointestinal pathobionts including Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum subsp. polymorphum, and Actinobacillus actinomycetemcomitans, Helicobacter pylori. Moreover, the postbiotic powder demonstrated gastroprotective effects by promoting recovery in a hydrogen peroxide-induced gastric injury model. Based on these findings, the postbiotic powder is safe, non-toxic, and suitable for oral consumption at the tested doses, with promising antimicrobial and gastroprotective potential. Future research should explore its potential applications in health promotion and food safety. Full article
(This article belongs to the Section Probiotic Strains and Fermentation)
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15 pages, 3966 KB  
Article
Preparation of Suaeda Tea Through Semi-Solid Fermentation Utilizing Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii: Physicochemical Characteristics, Process Optimization, and Antioxidant Activity
by Aoqi Dong, Xiaoying Dong, Xinying Dai, Yanru Gao, Yuewen Ning, Xiya Fan and Haiyan Liu
BioTech 2025, 14(4), 83; https://doi.org/10.3390/biotech14040083 - 28 Oct 2025
Viewed by 72
Abstract
Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic [...] Read more.
Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic blend of microbial agents: Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii. The resulting tea demonstrates a potent antioxidant capacity, with a hydroxyl radical scavenging rate of 64.2% and an exceptional 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity of 83.3%, along with increased ferric ion reduction/antioxidant power (FRAP) reducing power (1.82), indicating its superior antioxidant profile. Through the comparison of different microbial strain combinations under varying process parameters such as fermentation temperature and duration, the experiment revealed that fermentation at 37 °C for 24 h results in the highest concentrations of tea polyphenols (TPs) (≥10.87 mg/mL) and free amino acids (26.32 mg/100 mL). The quality of the fermented Suaeda tea meets the stringent GB/T 21733-2008 standards for tea beverages, exhibiting excellent physicochemical indices and sensory attributes. The antioxidant efficacy of the fermented Suaeda tea persists significantly throughout a 180-day duration. The optimization of the fermentation process for Suaeda tea not only provides a theoretical framework for large-scale production but also establishes a foundation for Suaeda salsa in the tea beverage sector. This innovation enriches the market with a diverse range of health-promoting teas, catering to the growing consumer demand for nutritious and beneficial beverages. Full article
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19 pages, 3786 KB  
Article
Metabolic Characterization of Two Flor Yeasts During Second Fermentation in the Bottle for Sparkling Wine Production
by Juan Carlos García-García, María Trinidad Alcalá-Jiménez, Juan Carlos Mauricio, Cristina Campos-Vázquez, Inés M. Santos-Dueñas, Juan Moreno and Teresa García-Martínez
Int. J. Mol. Sci. 2025, 26(21), 10457; https://doi.org/10.3390/ijms262110457 - 28 Oct 2025
Viewed by 222
Abstract
The global sparkling wine market continues to grow steadily, reaching approximately 24 million hectoliters in 2023, with an annual increase of around 4% despite a general decline in overall alcoholic beverage consumption. This growth highlights the importance of employing diverse yeast strains to [...] Read more.
The global sparkling wine market continues to grow steadily, reaching approximately 24 million hectoliters in 2023, with an annual increase of around 4% despite a general decline in overall alcoholic beverage consumption. This growth highlights the importance of employing diverse yeast strains to improve product variety and quality. Flor yeasts are specialized strains of Saccharomyces cerevisiae that develop a biofilm on the surface of certain wines during biological ageing. They possess unique physiological properties, including high ethanol tolerance and the capacity to adhere, which supports wine clarification. They also have the ability to contribute unique volatile compounds and aroma profiles, making them promising candidates for sparkling wine production. This study evaluated two flor yeast strains (G1 and N62), which were isolated from the Pérez Barquero winery during the second fermentation process using the traditional method. Sparkling wines were produced by inoculating base wine (BW) with each strain, and the wines were monitored at 3 bar CO2 pressure and after 9 months of ageing on lees. Comprehensive metabolomic analysis was performed using GC-MS for volatile compounds and HPLC for nitrogen compounds, with statistical analysis including PCA, ANOVA, Fisher’s LSD, and correction FDR tests. Strain N62 demonstrated faster fermentation kinetics and higher cellular concentration, reaching 3 bar pressure in 27 days compared to 52 days for strain G1. Both strains achieved similar final pressures, 5.1–5.4 bars. Metabolomic profiling revealed significant differences in the profiles of volatile and nitrogen compounds between the two strains. G1 produced higher concentrations of 3-methyl-1-butanol, 2-methyl-1-butanol, and acetaldehyde, while N62 generated elevated levels of glycerol, ethyl esters, and amino acids, including glutamic acid, aspartic acid, and alanine. These findings demonstrate that both flor yeast strains successfully complete sparkling wine fermentation while producing distinct metabolic signatures that could contribute to unique sensory characteristics. This supports their potential as alternatives to conventional sparkling wine yeasts for enhanced product diversification. Full article
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18 pages, 1729 KB  
Article
Sustainable 2-Phenylethanol Production: Co-Cultivation of Yarrowia lipolytica Strains in Mixed Agro-Industrial By-Products
by Sara Mitri, Nicolas Louka, Tristan Rossignol, Richard G. Maroun and Mohamed Koubaa
Fermentation 2025, 11(11), 611; https://doi.org/10.3390/fermentation11110611 - 28 Oct 2025
Viewed by 305
Abstract
The bioproduction of 2-phenylethanol (2-PE), a high-value aromatic compound widely used in the fragrance, cosmetic, food and beverage, and pharmaceutical industries, through yeast fermentation offers a sustainable alternative to chemical synthesis and rose extraction. This study explores the fermentation of Yarrowia lipolytica strains [...] Read more.
The bioproduction of 2-phenylethanol (2-PE), a high-value aromatic compound widely used in the fragrance, cosmetic, food and beverage, and pharmaceutical industries, through yeast fermentation offers a sustainable alternative to chemical synthesis and rose extraction. This study explores the fermentation of Yarrowia lipolytica strains using mixed agro-industrial by-products as substrates to produce 2-PE via de novo synthesis, without supplementation with the costly precursor L-phenylalanine. Y. lipolytica strains were genetically engineered to enhance flux through the shikimate pathway and enable the hydrolysis of a broader range of substrates. The culture media consisted solely of a mixture of agro-industrial by-products: sugar beet molasses (SBM), brewer’s spent grain (BSG) pressing extract, and chicory root (CR) pressing extract, serving as the primary carbon and nitrogen sources without the addition of nutrients, minerals, synthetic, complex ingredients, or costly additives. The co-culture approach enhanced substrate utilization, leading to an increase in 2-PE titers, reaching approximately 2.5 g/L 2-PE production after 240 h of fermentation. This study demonstrates the feasibility of integrating co-culture fermentation and agro-industrial waste valorization for sustainable 2-PE production, offering a scalable bioprocess for industrial applications. Full article
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18 pages, 1619 KB  
Article
Ferulic Acid Esterase-Producing Inoculant Improves Fiber Degradation and Modulates Microbial Diversity in Corn Bran Silage and Whole-Plant Corn Silage
by Yang Yu, Xiaojun Guo, Haoer Li, Chen Yu, Hao Liu and Wei Guo
Microorganisms 2025, 13(11), 2439; https://doi.org/10.3390/microorganisms13112439 - 24 Oct 2025
Viewed by 263
Abstract
Ferulic acid esterase (FAE) catalyzes the hydrolysis of the feruloyl ester bond in lignocellulose, exposing cellulose. The objective of this research was to examine the impacts of Bacillus amyloliquefaciens A30 producing FAE on the fermentation quality, fiber degradation, enzyme activity and microbial diversity [...] Read more.
Ferulic acid esterase (FAE) catalyzes the hydrolysis of the feruloyl ester bond in lignocellulose, exposing cellulose. The objective of this research was to examine the impacts of Bacillus amyloliquefaciens A30 producing FAE on the fermentation quality, fiber degradation, enzyme activity and microbial diversity of corn bran silage and whole-plant corn silage. The experimental treatments were as follows: control (CK), cellulase (CEL), strain A30 (A30) and CEL + A30. Corn bran and whole-plant corn were ensiled for 14 d and 60 d, respectively. The results showed that all additive treatments effectively reduced the pH, neutral detergent fiber, acid detergent fiber and cellulose contents of both corn bran silage and whole-plant corn silage in comparison with control, with CEL + A30 group performing the best effects. Meanwhile, higher FAE activity was detected in A30 and CEL + A30 groups during ensiling. Furthermore, the supplementation of A30 increased the degradation ratio of NDF, ADF, ADL, and cellulose of corn bran silage and whole-plant corn silage. Additionally, treatments with A30 and CEL + A30 increased the abundance of Lactobacillus, and reduced the proportion of pathogenic genera, including Acinetobacter, Enterobacter, and Sphingobacterium. In conclusion, the application of A30 may effectively promote fiber degradation and the stability of microecological system for corn silage. Full article
(This article belongs to the Section Microbial Biotechnology)
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26 pages, 663 KB  
Article
Probiotic Sheep Milk: Physicochemical Properties of Fermented Milk and Viability of Bacteria Under Simulated Gastrointestinal Conditions
by Małgorzata Pawlos, Katarzyna Szajnar and Agata Znamirowska-Piotrowska
Nutrients 2025, 17(21), 3340; https://doi.org/10.3390/nu17213340 - 24 Oct 2025
Viewed by 335
Abstract
Background/Objectives: Within the spectrum of lactic acid bacteria, Lacticaseibacillus casei and Lactobacillus johnsonii are of particular technological and nutritional significance. Protein fortification of fermented dairy systems offers dual benefits: it improves product quality while enhancing probiotic resilience. Supplementary proteins supply bioavailable nitrogen and [...] Read more.
Background/Objectives: Within the spectrum of lactic acid bacteria, Lacticaseibacillus casei and Lactobacillus johnsonii are of particular technological and nutritional significance. Protein fortification of fermented dairy systems offers dual benefits: it improves product quality while enhancing probiotic resilience. Supplementary proteins supply bioavailable nitrogen and peptides that stimulate bacterial metabolism and contribute to a viscoelastic gel matrix that buffers cells against gastric acidity and bile salts. The aim of this study was to clarify the functional potential of such formulations by assessing probiotic survival under in vitro digestion simulating oral, gastric, and intestinal phases. Methods: Sheep milk was fermented with L. casei 431 or L. johnsonii LJ in the presence of whey protein isolate (WPI), soy protein isolate (SPI), or pea protein isolate (PPI) at concentrations of 1.5% and 3.0%. Physicochemical parameters (pH, titratable acidity, color, syneresis), organoleptic properties, and microbiological counts were evaluated. The viability of L. casei and L. johnsonii was determined at each digestion stage, and probiotic survival rates were calculated. Results: Samples with L. johnsonii consistently exhibited lower pH values compared to L. casei. Across both bacterial strains, the addition of 1.5% protein isolate more effectively limited syneresis than 3.0%, regardless of protein type. Samples fortified with WPI at 1.5% (JW1.5) and 3.0% (JW3.0) were rated highest by the panel, demonstrating smooth, homogeneous textures without grittiness. The greatest bacterial survival (>70%) was observed in WPI-fortified samples (JW1.5, JW3.0) and in SPI-fortified JS3. Conclusions: Protein isolates of diverse origins are suitable for the enrichment of fermented sheep milk, with 1.5% supplementation proving optimal. Such formulations maintained desirable fermentation dynamics and, in most cases, significantly improved the survival of L. casei and L. johnsonii under simulated gastrointestinal conditions, underscoring their potential in the development of functional probiotic dairy products. Full article
(This article belongs to the Special Issue Probiotics, Postbiotics, Gut Microbiota and Gastrointestinal Health)
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Article
In Vitro Gastrointestinal Digestion of Grifola frondosa Polysaccharides and Their Enhancement of GABA Production via Gut Microbiota Modulation
by Qingchi Wang, Yuhang Luo, Huabo Zhu, Xiaoyang Liu, Mingyuan Xue, Guiling Yang, Yue Chen, Shiguo Chen and Zhengshun Wen
Nutrients 2025, 17(21), 3332; https://doi.org/10.3390/nu17213332 - 23 Oct 2025
Viewed by 202
Abstract
Background: The water-soluble Grifola frondosa polysaccharides (GFPs) are the primary bioactive component of the edible and medicinal fungus Grifola frondosa. However, the digestive behavior of GFPs in the human gastrointestinal (GI) tract and their subsequent interaction with gut microbiota (GM) to [...] Read more.
Background: The water-soluble Grifola frondosa polysaccharides (GFPs) are the primary bioactive component of the edible and medicinal fungus Grifola frondosa. However, the digestive behavior of GFPs in the human gastrointestinal (GI) tract and their subsequent interaction with gut microbiota (GM) to exert health effects remain unclear. Methods: In this study, GFPs were extracted based on a traditional hot water decoction. An in vitro simulated GI digestion model and a human fecal microbiota fermentation model were established to systematically investigate the digestive stability of GFPs, GM modulation, and metabolite changes. Results: Results showed that GFPs remained structurally stable during in vitro oral, gastric, and small intestinal digestion, allowing them to reach the colon intact for microbial fermentation. During colonic fermentation, GFPs were efficiently degraded by GM, and significantly increased the relative abundance of beneficial bacteria such as Akkermansia, Bacteroides, Parabacteroides, and Lactobacillus while reducing the abundance of pathogenic Escherichia-Shigella. Meanwhile, GFPs enriched metabolites beneficial for intestinal health, among which γ-aminobutyric acid (GABA) was the most significantly upregulated. Single-strain fermentation confirmed that Lactobacillus (L. plantarum) was the core GABA-producing genus. Conclusions: This study highlights the potential of GFPs as prebiotics for GM modulation, expands the understanding of the health-promoting effects of fungal polysaccharides, and provides a theoretical basis for the development of GFP-based functional foods. Full article
(This article belongs to the Section Prebiotics and Probiotics)
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