Fermentation

18 pages, 1973 KiB

Open AccessArticle

Potential of Whey Protein-Fortified Blackberry Juice in Transporting and Protecting Lactic Acid Bacteria: A Proteolytic Profile Analysis and Antioxidant Activity

by Liliana Lugo-Zarate, Angélica Saraí Jiménez-Osorio, Luis Guillermo González-Olivares, Emmanuel Pérez-Escalante, Araceli Castañeda-Ovando, José Pedraza-Chaverri, Omar Noel Medina-Campos, María Guadalupe Herrera-Hernández and Luis Delgado-Olivares

Fermentation 2025, 11(5), 252; https://doi.org/10.3390/fermentation11050252 - 2 May 2025

Abstract

This study investigates the potential of blackberry juice fortified with whey as a carrier for transporting and protecting lactic acid bacteria (LAB). The interactions between whey proteins and the juice were examined to assess their impact on probiotic stability and protection during storage [...] Read more.

This study investigates the potential of blackberry juice fortified with whey as a carrier for transporting and protecting lactic acid bacteria (LAB). The interactions between whey proteins and the juice were examined to assess their impact on probiotic stability and protection during storage and passage through the gastrointestinal tract. Additionally, the study explored how this combination influences the antioxidant properties of the product. The results indicated that the blackberry juice and whey protein mixture provided moderate protection to Lacticaseibacillus rhamnosus GG compared to the positive control (inulin), suggesting that whey proteins may enhance probiotic viability. Proteolytic analysis revealed progressive protein hydrolysis during fermentation, leading to the release of bioactive peptides, indicating the formation of compounds with potential functional benefits. Moreover, samples inoculated with LAB exhibited higher antioxidant activity than those without inoculum. This research demonstrates the promise of fermented blackberry juice fortified with whey proteins as an effective probiotic delivery system. It opens new possibilities for developing functional foods to promote intestinal health and overall well-being. Full article

(This article belongs to the Special Issue Lactic Acid Bacteria Metabolism)

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17 pages, 2461 KiB

Open AccessArticle

Optimization of Palm Kernel Cake Bioconversion with P. ostreatus: An Efficient Lignocellulosic Biomass Value-Adding Process for Ruminant Feed

by Aldo Ibarra-Rondón, Dinary Eloisa Durán-Sequeda, Andrea Carolina Castro-Pacheco, Pedro Fragoso-Castilla, Rolando Barahona-Rosales and José Edwin Mojica-Rodríguez

Fermentation 2025, 11(5), 251; https://doi.org/10.3390/fermentation11050251 - 1 May 2025

Abstract

This study aims to optimize the bioconversion of palm kernel cake (PKC) by Pleurotus ostreatus to improve fungal biomass production, lignocellulolytic enzyme expression, and the nutritional value of the substrate as ruminant feed. Three inorganic nitrogen sources (ammonium sulfate, ammonium nitrate, and urea) [...] Read more.

This study aims to optimize the bioconversion of palm kernel cake (PKC) by Pleurotus ostreatus to improve fungal biomass production, lignocellulolytic enzyme expression, and the nutritional value of the substrate as ruminant feed. Three inorganic nitrogen sources (ammonium sulfate, ammonium nitrate, and urea) were evaluated for fungal biomass production using a central composite design (CCD) in liquid fermentations. The formulated culture medium (18.72 g/L glucose and 0.39 g/L urea) effectively yielded better fungal biomass production (8 g/L). Based on these results, an extreme vertex design, mixtures with oil palm by-products (PK, hull, and fiber) supplemented with urea, were formulated, finding that PKC stimulated the highest biomass production and laccase enzyme activity in P. ostreatus. The transcriptome of P. ostreatus was obtained, and the chemical composition of the fermented PKC was determined. Transcriptomic analysis revealed the frequency of five key domains with carbohydrate-activated enzyme (CAZy) function: GH3, GH18, CBM1, AA1, and AA5, with activities on lignocellulose. In the fermented PKC, lignin was reduced by 46.9%, and protein was increased by 69.8%. In conclusion, these results show that urea is efficient in the bioconversion of PKC with P. ostreatus as a supplement for ruminants. Full article

(This article belongs to the Special Issue Fermentation Strategies to Enhance Feed Nutritional Value and Optimize Industry Resources)

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22 pages, 2120 KiB

Open AccessArticle

The Effect of Skin Contact, β-Lyase and Fermentation Gradient Temperature on Fermentation Esters and Free Volatile Thiols in Oregon Chardonnay Wine

by Angelica Iobbi, James Osborne, Yanming Di and Elizabeth Tomasino

Fermentation 2025, 11(5), 250; https://doi.org/10.3390/fermentation11050250 - 1 May 2025

Abstract

This study investigated specific winemaking procedures that could increase fermentation esters and volatile thiols in Chardonnay wine during fermentation. These compounds together are known to cause tropical fruit aromas. Two levels of pre-fermentative skin contact (10 °C for 18 h) (yes/no), two levels [...] Read more.

This study investigated specific winemaking procedures that could increase fermentation esters and volatile thiols in Chardonnay wine during fermentation. These compounds together are known to cause tropical fruit aromas. Two levels of pre-fermentative skin contact (10 °C for 18 h) (yes/no), two levels of β-lyase addition (40 μL/L) (yes/no), and three levels of fermentation gradient temperature, FG0 (constant 13 °C), FG1 (started at 20 °C and after 96 h dropped to 13 °C), and FG2 (started at 20 °C and after ~11.5 °Brix dropped to 13 °C), were evaluated using laboratory-scale ferments in a full factorial design. Esters and the volatile thiols, 3-sulfanylhexan-1-ol (3SH), 3-sulfanylhexyl acetate (3SHA), and 4-methyl-4-sulfanylpentan-2-one (4MSP), were quantified using gas and liquid chromatography methods, respectively. The combination of skin contact and FG1 or FG2 resulted in the greatest levels of esters and thiols in Chardonnay wine. The fermentation gradient was shown to be efficient in reducing volatile compounds normally lost due to evaporation during fermentation. With these different processing techniques, it will be possible for winemakers to achieve different wine qualities depending on their chosen wine style. Full article

(This article belongs to the Special Issue Alcoholic Fermentation)

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13 pages, 1633 KiB

Open AccessArticle

Metabolic Engineering for the Biosynthesis of Pentalenene in the Rapidly Growing Bacterium Vibrio natriegens

by Lujun Hu, Rui Lin, Hui Jiang, Ge Yao, Jiajia Liu, Penggang Han, Xiukun Wan, Chang Chen, Yunfei Zhang, Shaoheng Bao and Fuli Wang

Fermentation 2025, 11(5), 249; https://doi.org/10.3390/fermentation11050249 - 1 May 2025

Abstract

Vibrio natriegens (V. natriegens) is an emerging synthetic biology chassis characterized by rapid growth, and its potential for the synthesis of sesquiterpenes (such as pentalenene) has not been developed. In this study, heterologous pentalenene biosynthesis was successfully established in V. natriegens via metabolic [...] Read more.

Vibrio natriegens (V. natriegens) is an emerging synthetic biology chassis characterized by rapid growth, and its potential for the synthesis of sesquiterpenes (such as pentalenene) has not been developed. In this study, heterologous pentalenene biosynthesis was successfully established in V. natriegens via metabolic engineering. The optimization of gene dosage and culture conditions led to an increase in pentalenene yield from 0.75 mg/L to 39.4 mg/L, representing the highest titer reported in V. natriegens to date, though still markedly lower than yields achieved in conventional microbial hosts. Transcriptome analysis demonstrated that the exogenous mevalonate (MVA) pathway effectively activated terpenoid precursor synthesis, as evidenced by the up-regulation of key pathway genes. However, the endogenous methylerythritol 4-phosphate (MEP) pathway remained inactive, and genes involved in oxidative phosphorylation, the pentose phosphate pathway, and thiamine biosynthesis were down-regulated, leading to limited availability of ATP, NADPH, and acetyl-CoA. Competition for cofactors, particularly NADPH, further constrained precursor supply and pathway efficiency. This study confirmed the potential of V. natriegens as a pentalenene production platform and revealed its metabolic bottleneck, providing a theoretical basis for subsequent engineering optimization. Full article

(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)

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22 pages, 2478 KiB

Open AccessArticle

Optimized Spirulina Fermentation with Lacticaseibacillus rhamnosus: Bioactive Properties and Pilot-Scale Validation

by Akif Emre Kavak, Didem Balkanlı, Osman Sagdıc, Akın Özdemir and Enes Dertli

Fermentation 2025, 11(5), 248; https://doi.org/10.3390/fermentation11050248 - 1 May 2025

Abstract

Sustainable bio-based products derived from fermentation are gaining increasing interest. The present study was designed to determine the interaction of Lacticaseibacillus rhamnosus 23.2 bacteria with spirulina in a 3 L glass bioreactor and the effect of aeration and agitation speed on the final [...] Read more.

Sustainable bio-based products derived from fermentation are gaining increasing interest. The present study was designed to determine the interaction of Lacticaseibacillus rhamnosus 23.2 bacteria with spirulina in a 3 L glass bioreactor and the effect of aeration and agitation speed on the final product biomass and antioxidant capacity. The fermentation medium contained only glucose, an inorganic salt mixture, and spirulina powder. The estimated biomass and antioxidant activity were found to be 3.74 g/L and 84.72%, respectively, from the results of the optimization model. Scale-up was performed with the obtained optimization data, and three pilot-scale fermentations were carried out in a 30 L stainless steel bioreactor. As a result of pilot production, the obtained bioactive products were freeze-dried, and their antibacterial, antioxidant, total phenolic properties, and cytotoxic activity were investigated. The pilot production results showed that the increase in bacterial cell number was around 3–4 log after 24 h of fermentation. An inhibitory effect against pathogenic bacteria was observed. A strong radical scavenging effect was found in antioxidant analyses. Total phenolic substance content was 26.5 mg gallic acid equivalent (GAE) g⁻¹, which was the highest level in this study. Cytotoxic activity showed that bioactive products had a cytotoxic effect against Caco-2 adenocarcinoma cells. This study emphasizes the potential of Arthrospira platensis biomass as a substrate for the production of lactic acid bacteria (LAB)-based bioproducts. It is thought that the results obtained from this study may position potential innovative strategies in the food, pharmaceutical, agriculture, and cosmetic industries. Full article

(This article belongs to the Special Issue 10th Anniversary of Fermentation: Feature Papers in Section "Industrial Fermentation")

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11 pages, 1850 KiB

Open AccessArticle

Effects of Different Levels of Thiamine Diphosphate on In Vitro Methane Reduction and Fermentation Characteristics of Korean Native Cow (Hanwoo)

by Seong-Shin Lee, Seong-Uk Jo, Heoyn-Sang Kim, Ji-Soo Wi, Yoo-Kyung Lee and Seong-Dae Lee

Fermentation 2025, 11(5), 247; https://doi.org/10.3390/fermentation11050247 - 1 May 2025

Abstract

This study investigated the effects of different doses of thiamine diphosphate (ThDP) on methane reduction and fermentation characteristics of Korean native cow (Hanwoo). In vitro trials used ThDP powder at 240, 360, 480, 600, and 720 ppm of substrate, with each sample incubated [...] Read more.

This study investigated the effects of different doses of thiamine diphosphate (ThDP) on methane reduction and fermentation characteristics of Korean native cow (Hanwoo). In vitro trials used ThDP powder at 240, 360, 480, 600, and 720 ppm of substrate, with each sample incubated at 39 °C for 24 and 48 h. After incubation, each sample was analyzed for total gas, methane production, dry matter digestibility, and rumen fermentation characteristics. Mean comparisons were performed using Tukey’s test, with significant differences declared at p < 0.05. Total gas production, methane ratio, and methane production per digested dry matter had a quadratic pattern (p < 0.001), and the 480 ppm treatment had the lowest (p < 0.05) at 24 and 48 h of incubation. Total volatile fatty acid concentration showed no significant difference at 24 h but differed significantly at 48 h (p > 0.05). The concentration of propionate had a quadratic pattern (p < 0.001), and the 480 ppm treatment showed the highest levels compared to the other treatments (p < 0.05) after 24 and 48 h of incubation. In conclusion, ThDP supplementation had a methane inhibition effect. In particular, the methane inhibition effect was most pronounced when ThDP was supplemented at 480 ppm. Full article

(This article belongs to the Special Issue Ruminal Fermentation)

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15 pages, 3914 KiB

Open AccessArticle

Efficient Synthesis of High-Active Myoglobin and Hemoglobin by Reconstructing the Mitochondrial Heme Synthetic Pathway in Engineered Saccharomyces cerevisiae

by Xiaoyan Sun, Yunpeng Wang, Yijie Wang, Jingwen Zhou, Jianghua Li, Jian Chen, Guocheng Du and Xinrui Zhao

Fermentation 2025, 11(5), 246; https://doi.org/10.3390/fermentation11050246 - 1 May 2025

Abstract

Currently, various types of myoglobins and hemoglobins are widely used in the fields of food additives and biocatalytic applications. However, the limited availability of heme constrains the biosynthesis of these high-activity hemoproteins in microbial chassis cells. In this work, a new heme synthetic [...] Read more.

Currently, various types of myoglobins and hemoglobins are widely used in the fields of food additives and biocatalytic applications. However, the limited availability of heme constrains the biosynthesis of these high-activity hemoproteins in microbial chassis cells. In this work, a new heme synthetic pathway was reconstructed in the mitochondria by eliminating the spatial barrier during heme synthesis in Saccharomyces cerevisiae, resulting in a significant enhancement in intracellular heme supply. To further enhance the supply of the essential precursor for heme synthesis (5-aminolevulinate, ALA), the special ALA exporter in the mitochondrial membrane (Ort1p) was identified and knocked out. Moreover, the mitochondrial heme exporter (Ygr127wp) was overexpressed to promote the transport of heme to the cytoplasm to participate in the synthesis of various myoglobins and hemoglobins. Based on these strategies in the engineered strain, the binding ratios of heme in porcine myoglobin (52.4 ± 4.9%) and soybean hemoglobin (75.5 ± 2.8%) were, respectively, increased by 2.4-fold and 3.6-fold, and the titers of porcine myoglobin (130.5 ± 2.8 mg·L⁻¹) and soybean hemoglobin (152.8 ± 2.6 mg·L⁻¹), respectively, increased by 31.1% and 42.1%. Furthermore, the engineered strain presents great potential in the efficient synthesis of other heme-binding proteins and enzymes in S. cerevisiae. Full article

(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)

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14 pages, 4818 KiB

Open AccessArticle

Utilizing Agrobyproducts: Potential Alternative Substrates for Cultivation of Lentinula edodes

by Zhiguo Zhou, Guohui Cheng, Wenjie Chen, Chunyan Xie, Guisen Hou and Xiaoya An

Fermentation 2025, 11(5), 245; https://doi.org/10.3390/fermentation11050245 - 30 Apr 2025

Abstract

This study evaluated six agrobyproducts (oak, jujube, apple, pear, peach, black locust) as alternative substrates for Lentinula edodes cultivation to mitigate oak dependency. Twelve substrate formulations were tested, including individual and mixed sawdust combinations. Results demonstrated successful mycelial colonization across all treatments, with [...] Read more.

This study evaluated six agrobyproducts (oak, jujube, apple, pear, peach, black locust) as alternative substrates for Lentinula edodes cultivation to mitigate oak dependency. Twelve substrate formulations were tested, including individual and mixed sawdust combinations. Results demonstrated successful mycelial colonization across all treatments, with treatment PAS (78% pear sawdust, 20% wheat bran, 1.5% gypsum, and 0.5% lime) exhibiting the fastest mycelial growth (4.70 mm/day) and full colonization in 105 days. Treatment BLS (78% black locust sawdust, 20% wheat bran, 1.5% gypsum, and 0.5% lime) achieved the highest biological efficiency (97.26%) and productivity (0.85 kg/bag). Nutrient analysis revealed substrate-specific enhancements: PAS maximized vitamin C (4.88 mg/100 g) and iron, while PAS + OS (39% peach sawdust, 39% oak sawdust, 20% wheat bran, 1.5% gypsum, and 0.5% lime) elevated protein (3.88%), phosphorus, and zinc. PCA highlighted distinct nutritional profiles for BLS- and jujube-based mushrooms. Correlation analyses identified the third (r = 0.838) and fourth flushes (r = 0.922) as critical for total yield, with selenium and zinc significantly linked to growth rates. Black locust and peach substrates outperformed or complemented oak, offering sustainable alternatives. These findings underscore the potential of agrowaste utilization to reduce ecological strain while maintaining high yields and nutritional quality, aligning with global agricultural sustainability goals. Full article

(This article belongs to the Section Industrial Fermentation)

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16 pages, 2167 KiB

Open AccessArticle

Growth Performance and Rumen Microbiota of Sheep Respond to Cotton Straw Fermented with Compound Probiotics

by Peiling Wei, Mingxuan Guan, Xuhui Liang, Kaixin Yuan, Ning Chen, Yuxin Yang and Ping Gong

Fermentation 2025, 11(5), 244; https://doi.org/10.3390/fermentation11050244 - 29 Apr 2025

Abstract

To develop cotton straw as a feed resource through biological fermentation, it was fermented using compound probiotics (Bacillus subtilis, Saccharomyces cerevisiae, and Lactobacillus plantarum) and subsequently fed to sheep after the nutrients and hygienic indices of the fermented cotton straw [...] Read more.

To develop cotton straw as a feed resource through biological fermentation, it was fermented using compound probiotics (Bacillus subtilis, Saccharomyces cerevisiae, and Lactobacillus plantarum) and subsequently fed to sheep after the nutrients and hygienic indices of the fermented cotton straw (FCS) were analyzed. Sixty sheep were randomly assigned to five groups: a control group (CON); a low-proportion fermented cotton straw group (LFC, with FCS comprising 14.5% of the diet); a high-proportion fermented cotton straw group (HFC, with FCS comprising 29.0% of the diet); a compound microbial group (MIC, containing Bacillus licheniformis, Bacillus subtilis, and yeast); and a microbial-enzymatic preparation group (MEY, containing compound probiotics and enzymes such as cellulase, xylanase, β-glucanase, amylase, and protease). The trial lasted seven weeks and was divided into two stages: stage 1 (weeks 1–4, days 1–28) and stage 2 (weeks 5–7, days 29–49). Body weight and daily feed intake were registered, and blood and rumen fluid samples were obtained at day 28 and day 49 of the feeding trial. Fermentation significantly increased the crude protein content of cotton straw while reducing neutral detergent fiber (NDF) and acid detergent fiber (ADF) (p < 0.05). Additionally, fermentation reduced the residues of aflatoxin B1, vomitoxin, zearalenone, and free gossypol in the treatment groups (p < 0.05). LFC possessed the lowest value of feed-to-gain ratio (F/G) among all groups. Serum indices related to antioxidant capacity and utilization of fat and protein increased in the treatment group (p < 0.05). Rumen microbiota were separated between different groups (p < 0.05). LFC and HFC enhanced the abundance of Prevotella. These findings could provide conclusions that fermented cotton straw has the tendency to enhance the growth performance of sheep by increasing the abundance of bacteria related to utilization of protein, carbohydrate, and other nutrients such as Prevotella, in which the LFC group has the best fast-fattening (about 50 d) effect. Full article

(This article belongs to the Special Issue Application of Fermentation Technology in Animal Nutrition: 2nd Edition)

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17 pages, 1990 KiB

Open AccessArticle

Hydrotalcites as a Promising Adsorbent for Hemicellulose Hydrolysate Detoxification in Xylitol Production

by Débora D. V. da Silva, Kelly J. Dussán, Isabela A. L. Costa, Marcus B. S. Forte and Maria G. A. Felipe

Fermentation 2025, 11(5), 243; https://doi.org/10.3390/fermentation11050243 - 27 Apr 2025

Abstract

The worldwide demand for sustainable bioprocesses is undeniable, as well as for research aimed at the biotechnological exploitation of lignocellulosic materials, especially their hemicellulosic fractions rich in xylose. Various bioproducts can be obtained from these fractions, although some bottlenecks still exist, such as [...] Read more.

The worldwide demand for sustainable bioprocesses is undeniable, as well as for research aimed at the biotechnological exploitation of lignocellulosic materials, especially their hemicellulosic fractions rich in xylose. Various bioproducts can be obtained from these fractions, although some bottlenecks still exist, such as the presence in hemicellulosic hydrolysates of compounds that are toxic for microorganisms, which requires a previous step of detoxification to reduce them to non-inhibitory levels. The present investigation proposes the use of hydrotalcites as a new detoxifying agent for the hemicellulosic hydrolysate of sugarcane straw to produce xylitol by Candida tropicalis, aiming at a greater removal of phenolics and less loss of sugars. The design of these experiments was used for factorial effect analysis in a simultaneous way; the influences of pH and temperature were evaluated, considering the detoxification process at different times for both uncalcined and calcined hydrotalcite adsorbents. While for the calcined hydrotalcite, the temperature was the significant factor, for the uncalcined, there was also an influence of pH and little effect on the factors of yield and productivity. The effectiveness of hydrotalcites as demonstrated in this research, mainly regarding the ability to reduce the content of phenolic compounds in hydrolysates with a low loss of sugar content, followed by fermentability to produce xylitol, is a strong requirement for the proposition of these new adsorbents in investigations of the development of sustainable technologies for obtaining bioproducts in a biorefinery context. Full article

(This article belongs to the Special Issue Bioprocesses for Biomass Valorization in Biorefineries)

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17 pages, 4633 KiB

Open AccessArticle

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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Graphical abstract

17 pages, 306 KiB

Open AccessReview

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

Open AccessArticle

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

Open AccessArticle

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 µ_x,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/g_cell. 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

Open AccessArticle

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

Open AccessArticle

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

Open AccessReview

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

Open AccessArticle

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

Open AccessReview

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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