Fermentation

Artisanal cheeses are known as the source of beneficial lactic acid bacteria (LAB). Therefore, this study aimed to isolate and characterize LAB with different proteolytic activities from Iranian artisanal white cheeses. The isolates were classified into low, medium, and high proteolytic activity clusters via K-means clustering and identified as Lactiplantibacillus (Lpb.) pentosus L11, Lpb. plantarum L33, and Enterococcus faecium L13, respectively. Some safety tests (such as resistance to antibiotics, hemolytic activity, and biogenic amine production), probiotic properties (including cell surface hydrophobicity, auto/co-aggregation, and antibacterial activity), and production of volatile compounds were evaluated. These were non-hemolytic and non-biogenic amine producers, and showed no irregular antibiotic resistance. Lpb. plantarum L33 had the highest hydrophobicity (30.55%) and auto-aggregation (49.56%), and the highest co-aggregation was observed for Lpb. pentosus L11 with Staphylococcus aureus (61.51%). The isolates also showed a remarkable antibacterial effect against pathogenic bacteria. Moreover, Lpb. pentosus L11 and Lpb. plantarum L33 with low and medium proteolytic activity produced a wider range of volatile compounds in milk compared to the strain with a high proteolytic effect. The results showed that a probiotic strain with low or medium proteolytic activity could improve the flavor characteristics of fermented milk. Full article

Changes in cultivation conditions, in particular salinity and temperature, affect the production of secondary fungal metabolites. In this work, the extracts of fungus previously described as Penicillium dimorphosporum cultivated in various salinity and temperature conditions were investigated using HPLC UV/MS techniques, and their DPPH radical scavenging and cytotoxicity activities against human prostate cancer PC-3 cells and rat cardiomyocytes H9c2 were tested. In total, 25 compounds, including 13 desoxyisoaustamide-related alkaloids and eight anthraquinones, were identified in the studied extracts and their relative amounts were estimated. The production of known neuroprotective alkaloids 5, 6 and other brevianamide alkaloids was increased in hypersaline and high-temperature conditions, and this may be an adaptation to extreme conditions. On the other hand, hyposalinity stress may induce the synthesis of unidentified antioxidants with low cytotoxicity that could be very interesting for future investigation. The study of secondary metabolites of the strain KMM 4689 showed that although brevianamide-related alkaloids and anthraquinone pigments are widely distributed in various fungi, these metabolites have not been described for P. dimorphosporum and related species. For this reason, the strain KMM 4689 was re-sequenced using the β-tubulin gene and ITS regions as molecular markers and further identified as P. hispanicum. Full article

FODMAPs are fermentable oligo-, di-, monosaccharides, and polyols. The application of homofermentative lactic acid bacteria (LAB) has been investigated as a promising approach for producing low-FODMAP whole-wheat bread. The low-FODMAP diet is recommended to treat irritable bowel syndrome (IBS). Wheat flour is staple to many diets and is a significant source of fructans, which are considered FODMAPs. The reduction of fructans via sourdough fermentation, generally associated with heterofermentative lactic acid bacteria (LAB), often leads to the accumulation of other FODMAPs. A collection of 244 wild-type LAB strains was isolated from different environments and their specific FODMAP utilisation profiles established. Three homofermentative strains were selected for production of whole-wheat sourdough bread. These were Lactiplantibacillus plantarum FST1.7 (FST1.7), Lacticaseibacillus paracasei R3 (R3), and Pediococcus pentosaceus RYE106 (RYE106). Carbohydrate levels in flour, sourdoughs (before and after 48 h fermentation), and resulting breads were analysed via HPAEC-PAD and compared with whole-wheat bread leavened with baker’s yeast. While strain R3 was the most efficient in FODMAP reduction, breads produced with all three test strains had FODMAP content below cut-off levels that would trigger IBS symptoms. Results of this study highlighted the potential of homofermentative LAB in producing low-FODMAP whole-wheat bread. Full article

Blueberries have received great attention due to the health effects of their bioactive compounds, such as antioxidant, antitumor, and anti-obesity properties. Probiotics also have these health-promoting benefits. However, these biological activities may be affected by the processs of gastrointestinal digestion, which decreases their functionality. This study aimed to use a more convenient method to improve the blueberries’ antioxidant activity and protective effects on probiotic cells by fermentation with whey protein, and to explore the possible mechanisms underlying these effects. This result showed that the total phenolic content, anthocyanin content, reducing power, DPPH radical scavenging capacity, and probiotic cells’ survival in a blueberry juice and whey protein fermentation system were enhanced in a model of in vitro gastrointestinal digestion. The bioactive compounds in blueberry juice interacted with whey protein, as shown through FTIR. The stability of phenolic compounds was enhanced, and the release of functional compounds in the mixture fermentation system was delayed through CLSM. Interactions between bioactive compounds in blueberries, whey protein, and bacterial surface proteins, glycoproteins or polysaccharides during fermentation were studied by SDS-PAGE. Thus, the stability of bioactive activities in the mixed system after fermentation was strengthened by the interaction. The mixed fermentation system has promising potential for improving antioxidant activity and protecting probiotic cells. Full article

This study aimed to obtain novel bioactive compounds derived from actinomycetes associated with marine biota from the coast of Indonesia. Actinomycetes have been identified as a potential source of bioactive compounds through enzymatic fermentation. In order to obtain bioactive compounds from the results of the biorefinery process, cultivation was performed by solid state fermentation (SSF) on shrimp shell waste medium. The inhibitory activity against pathogenic microorganisms was measured based on the optical density of samples from Gorontalo and Buleleng, Bali, Indonesia. Six isolates had a clear zone as an indicator of the chitinase activity of chitinase enzymes. The SSF extract, obtained after 14 days of incubation, was assayed for its antimicrobial activity by the 96-well plate microtiter method. Among the six isolates, isolate 19B19A1 had antibacterial and antifungal activity against Staphylococcus aureus and Malassezia globosa, respectively. DNA analysis indicated that the 19B19A1 isolate was Streptomyces tritolerans 19B19A1 and that its extract contained an alkaloid component that played a role in antimicrobial activity. These results indicate that shrimp shells can be used as a nutrient-rich alternative culture medium for actinomycetes. This study is expected to become a source of information related to biorefinery, especially in the exploration of bioactive compounds produced by actinomycetes. Full article

Lentinan has important applications in the food and medicine fields. Fermenting Lentinula edodes with Bacillus subtilis natto increased the lentinan extraction yield by 87.13% and greatly altered the molecular structure and antioxidant activity of lentinan. The uronic acid content in the lentinan molecular structure increased from 2.08% to 4.33%. The fermentation process did not affect the monosaccharide composition of lentinan, comprised of more than 90% glucose residues. Fermentation significantly reduced the molecular weight of lentinan and altered its apparent structure. The water solubility of fermented lentinan was increased by 165.07%, and the antioxidant activity was significantly improved. Fermentation using soybean as a substrate may be beneficial for enhancing the activity of Bacillus subtilis natto and producing lentinan with different molecular weights. Full article

Green methods of synthesizing nanoparticles are safer than chemical and physical methods, as well as being eco-friendly and cost-efficient. In this study, we use copper oxide nanoparticles (CuO NPs) fabricated with Sesbania grandiflora (Sg) (Hummingbird tree) leaves to test the effectiveness of green synthesizing methods. The attained Sg-CuO NPs physical and optical nature is characterized by UV-Vis spectroscopy Differential Reflectance Spectroscopy (UV-Vis DRS), Fourier Transform Infra-Red spectroscopy (FTIR), X-ray Diffraction spectroscopy (XRD), Scanning Electron Microscope (SEM), and Energy Dispersive X-ray Analysis (EDAX). UV-Vis spectrum for Sg-CuO NPs revealed a peak at 410 nm. SEM images showed the aggregation of needle-shaped particles, at a size of 33 nm. The amylase and glucosidase enzymes were inhibited by the Sg-CuO NPs up to 76.7% and 72.1%, respectively, indicating a possible antihyperglycemic effect. Fabricated Sg-CuO NPs disclosed the excellent inhibition of DPPH-free radicle formation (89.7%) and repressed protein degradation (81.3%). The results showed that Sg-CuO NPs display good anti-bacterial activity against the gram-negative (Escherichia coli and Pseudomonas aeruginosa) and gram-positive (Staphylococcus aureus). Cytotoxicity of the Sg-CuO NPs was determined using anIC₅₀ of 37 μg/mL. Sg-CuO NPs have shown promising anti-diabetic, anti-oxidant, protein degradation-inhibiting, and anti-microbial properties. Our findings have shown that synthesized Sg-CuO NPs have biological activities that may be utilized to treat bacterial infections linked to hyperglycemia. Full article

Corn stover (CS) is an abundant lignocellulosic by-product of the grain industry. Ferulic acid esterase producing (FAE+)-lactobacilli can potentially improve ensiled forages’ nutritive value through the hydrolysis of ferulic acid ester bonds present in cell walls during the fermentation process, but this has not been addressed in CS silage. In this study, we characterized 8 FAE+ lactobacilli regarding their FAE activity and inoculant aptitude: Lactobacillus (L.) johnsonii (CRL2237, CRL2238, CRL2240), L. plantarum (ETC182, CRL046, CRL2241), L. fermentum CRL1446 and L. brevis CRL2239. Next, 25% dry matter (DM) CS mini silos were prepared and either not inoculated (UN) or inoculated with each strain (10⁵ CFU g fresh matter⁻¹). Compared to UN, DM loss was significantly reduced in CRL046 and CRL2239, and organic matter increased in CRL2241-inoculated silages. Although the rest of the digestibility measures were not improved, in situ acid detergent fiber degradability (ADFD) was increased by the CRL2238 strain when compared to UN. Results in inoculated silages were not correlated with FAE activity quantification or growth/acidification studies in a CS-derived culture broth. This study demonstrates the potential of several FAE+ lactobacilli strains as CS inoculants and encourages further research. Full article

Pickles are a type of traditional fermented vegetables in China. To ascertain the effect of different lactic acid bacteria on pickles, the chemical composition characteristics, flavor substances, and bacterial diversity of the pickles fermented by natural bacteria, Lactobacillus plantarum R5, Lactobacillus pentosus R8, and L. plantarum R5 plus L. pentosus R8 were investigated in this study. The results showed that Lactobacillus enhanced the decrease in pH, increase in total acid content, degradation of nitrite, and production of organic acid (lactic acid and malic acid) of fermented pickles. A total of 80 flavors were detected in the pickles fermented for 14 days, and esters in pickles fermented by Lactobacillus were more plentiful. Firmicutes emerged as the predominant microbial phyla. Amongst these, the commonly encountered microorganisms were Lactobacillus, unclassified Enterobacteriaceae, Pantoea, and Weissella. The multivariate statistical analysis further showed that Lactobacillus had a strong negative correlation with pH and a strong positive correlation with malic acid and lactic acid, and the microorganisms in pickles could acclimate to the changing fermentation environment. The insights gained from this study may be of assistance to us in obtaining new insights into the microbiota succession and chemical compounds involved in the pickles fermented by Lactobacillus. Full article

DNA markers help detect the intraspecific genetic diversity of yeast strains. Eight ISSR (Inter Simple Sequence Repeats) primers were used to assess the intraspecific diversity of Saccharomyces cerevisiae (n = 96) from different populations (n = 3), evaluate the technological characteristics, and investigate trait-loci associations. The primers amplified 154 reproducible and scorable bands, of which 79.87% were polymorphic. The UPGMA (unweighted pair group method with arithmetic mean) dendrogram clustered 96 isolates into two main clusters, supported by STRUCTURE HARVESTER results (ΔK = 2). Analysis of molecular variance (AMOVA) indicated significant genetic differences between (15%) and within the populations (85%) (p < 0.001). Twenty-nine genetically distinct strains were selected for the technological characterization. Principal component analysis (PCA) revealed that five strains with high fermentation capacity, leavening activity, high growth index at 37 °C, and harsh growth conditions were technologically relevant. Trait-loci association analyses indicated that the highest correlation (r = 0.60) was recorded for the fermentation capacity on the 8th and 113th loci, amplified by ISSR-1 and ISSR-6 primers, respectively (p < 0.05). The strains yielding high performances and the associated loci amplified by ISSR markers possess a high potential to generate locus-specific primers to target the strains with high fermentation capacity. Full article

Food acceptability and appeal are significantly influenced by colour. Harmful effects associated with synthetic colorants are well established, and research is currently focused on developing natural, synthetic chemical-free substitutes from fungal sources, with broad applications in food, medicine, textiles and agriculture. Additionally, the market’s dearth of natural red colour substitutes requires the creation of novel red pigment alternatives from secure and scalable sources. The goal of the current research was to establish new endophytic marine fungi that are naturally occurring bio-sources of the red pigment. Based on its profuse extracellular red pigment-producing capacity, the fungus CPEF02 was selected and identified as Monascus purpureus CPEF02 via internal transcribed spacer (ITS) sequences and phylogenetic analysis. The chemical moieties of the pigmented extracts were identified by liquid chromatography-high resolution mass spectrometry (LC-HRMS). The optimal culture conditions for maximum pigment production were investigated by surveying various media compositions. The methanolic fungal colourant extract was shown to have substantial antibacterial and antifungal activities against anthropogenic pathogens, Staphylococcus aureus (MTCC 1430), methicillin-resistant Staphylococcus aureus (ATCCBAA811), Salmonella typhimurium (MTCC 3241) and Vibrio cholerae (N16961) at a 100 µg/mL concentration and at a 1 mg/mL concentration for Alternaria solani (ITCC 4632) and Rhizoctonia solani (AG1-IA). This extract also exhibited antioxidant activity against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical with an IC₅₀ of 14.42 µg/mL and a Trolox equivalent antioxidant capacity of 0.571 µM Trolox/µg of the methanolic colourant extract. The findings suggested that M. purpureus’s pigment could be a source of an industrially useful natural red colourant. Full article

The biodiversity of some Saccharomyces (S.) strains for fermentative activity and metabolic capacities is an important research area in brewing technology. Yeast metabolism can render simple beers very elaborate. In this review, we examine much research addressed to the study of how different yeast strains can influence aroma by chemically interacting with specific aromatic compounds (mainly terpenes) from the hop. These reactions are commonly referred to as biotransformations. Exploiting biotransformations to increase the product’s aroma and use less hop goes exactly in the direction of higher sustainability of the brewing process, as the hop generally represents the highest part of the raw materials cost, and its reduction allows to diminish its environmental impact. Full article

The use of alternative ingredients in the production of kombucha has seen a recent increase. Our research aimed to characterize the chemical, nutritional, microbial, and aromatic profiles of kombucha beverages prepared with strawberry tree (Arbutus unedo) fruits fermented with three different SCOBYs for 21 days. The analyses showed similar levels of microbiological groups (aerobic mesophilic microorganisms, lactic acid bacteria, acetic acid bacteria, and yeasts)among the SCOBYs used. The beverages studied displayed a decrease in pH value and carbohydrate content, and protein degradation was also observed as fermentation progressed. However, the increase in total phenolic compounds during the first week proved to be a point of interest. A total of 20 volatile organic compounds were detected, giving different sensory qualities to the beverages: higher ethanol, benzaldehyde-4-ethyl, or acetic acid depending on the SCOBY used. The results obtained indicated that strawberry tree kombucha might be an alternative beverage with notable nutritional and aromatic properties, with fermentation time and SCOBY composition being identified as crucial factors. Full article

Nowadays, finding out new natural scaffolds of microbial origin increases at a higher rate than in the past decades and represents an auspicious route for reinvigorating the pool of compounds entering pharmaceutical industries. Fungi serve as a depository of fascinating, structurally unique metabolites with considerable therapeutic significance. Aspergillus genus represents one of the most prolific genera of filamentous fungi. Aspergillus nidulans Winter G. is a well-known and plentiful source of bioactive metabolites with abundant structural diversity, including terpenoids, benzophenones, sterols, alkaloids, xanthones, and polyketides, many of which display various bioactivities, such as cytotoxicity, antioxidant, anti-inflammatory, antiviral, and antimicrobial activities. The current work is targeted to survey the reported literature on A. nidulans, particularly its metabolites, biosynthesis, and bioactivities, in addition to recent reports on its biotechnological potential. From 1953 till November 2022, relying on the stated data, 206 metabolites were listed, with more than 100 references. Full article

In this study, the effects of the fermentation kinetics, determination of the number of lactic acid bacteria, texture, water holding capacity, and color of fermented soy beverages with acai powder (3 and 6% w/v) were investigated. The addition of acai powder significantly influenced the fermentation kinetics based on changes in pH, accelerating fermentation in the initial period. The results showed that the acai additive did not affect the enumeration of Lactobacillus acidophilus and Bifidobacterium animalis subsp. lactis. The presence of acai inhibited the proliferation of Streptococcus thermophilus compared to the soy beverage without acai powder added. However, the higher the acai additive, the more Streptococcus thermophilus bacteria were detected: 4.39 CFU/g for 6% acai powder sample and 3.40 CFU/g for 3% acai powder sample. The addition of acai to the soy beverage reduced its firmness, consistency, cohesiveness, and viscosity index after fermentation. A slight difference was observed in the lightness and whiteness of fermented soy beverages with 3% and 6% acai powder. Full article

During aerobic exposure of silage, the fatty acid and amino acid composition may alter the quality and palatability, resulting in economic losses in livestock production. The objective of this study was to evaluate the effects of Lactiplantibacillus plantarum (LP), Lenti Lentilactobacillus buchneri (LB), and a mixture of LP and LB (PB) on the fatty acids, amino acids, and antioxidant capacity of Leymus chinensis silage during aerobic exposure. The lactic acid bacteria were added at 1 × 10⁶ CFU/g. The silage treatments were opened after 60 days of fermentation, and sampled on days 0, 4, and 8 of aerobic exposure. The LB group had higher total fatty acid and polyunsaturated fatty acid content, and less decrease in amino acid content and antioxidant capacity, while the LP group had a higher monounsaturated fatty acid content but a larger decrease in all indicators after exposure. Correlation analysis showed that Lactobacillus, Cryptococcus, Penicillium, and Thermoascus were more correlated with fatty acid changes, and that Lactobacillus, Actinomyces, Clostridium, and Penicillium were more correlated with amino acid changes. In conclusion, Lentilactobacillus buchneri could effectively improve the antioxidant capacity and fatty acid and amino acid contents of Leymus chinensis silage during aerobic exposure, while Lactiplantibacillus plantarum could effectively improve the content of each index of Leymus chinensis silage at opening, but deterioration was faster during aerobic exposure. Full article

Soybean meal is a class of by-products obtained from the processing of soybean products. Despite its high nutritional value, the presence of glycoside isoflavones limits human use of soybean meal. This study evaluated the effect of solid-state fermentation (SSF) with different edible mushroom mycelia (Pleurotus ostreatus, Hericium erinaceus, and Flammulina velutipes) on the proximate composition, antioxidant properties, and physicochemical properties of fermented soybean meal powder (SP). The results revealed that fermented SP had a higher nutritional value when compared to SP. P. ostreatus was the most pronounced among the three species. Crude protein content was found to have increased by 9.49%, while the concentration of glutamate and aspartic acid increased by 23.39% and 23.16%, respectively. SSF process significantly increased the total polyphenol content (TPC) and aglycone isoflavone content by 235.9% and 324.12%, respectively, resulting in increased antioxidant activity (evaluated by the DPPH, •OH, ABTS⁺ assays). Microstructural changes in fermented SP and nutrient degradation and utilization were observed. Thus, fermented SP can be used as a raw material with enhanced nutritional properties to develop new functional foods, such as plant-based foods represented by plant meat. It provides a promising approach for increasing the added value of soybean meal. Full article

As an anaerobic butyrate-producing bacterium, Clostridium cellulovorans can secrete a variety of extracellular enzymes to degrade plant-based cellulose. However, with glucose as the carbon source, it still secretes a large amount of protein in the broth. The metabolism and regulation are obscure and need to be further studied. Hence, in this study, C. cellulovorans was used to conduct fed-batch fermentation of glucose and microcrystalline at pH 7.0 to produce a higher level of butyrate in the bioreactor. It produced 16.8 mM lactate, 22.3 mM acetate, and 132.7 mM butyrate in 72 h during glucose fermentation. In contrast, it produced only 11.5 mM acetate and 93.9 mM butyrate and took 192 h to complete the fermentation with cellulose as the carbon source. Furthermore, there was no lactate detected in the broth. The analysis of carbon source balance and redox balance showed that 57% of the glucose was consumed to form acids in glucose fermentation, while only 47% of the cellulose was used for acid generation in the cellulose fermentation. Meanwhile, a large amount of protein was detected in the fermentation broth in both glucose (0.9 ± 0.1 g/L) and cellulose (1.1 ± 0.2 g/L) fermentation. These results showed that protein was also a main product. C. cellulovorans metabolized glucose to generate intermediate metabolites and reducing powers (NADH and Fd_red), then protein and acid synthesis consumed this reducing power to maintain the carbon source balance and redox balance in the cell metabolism. The results of comparative transcriptomics and comparative proteomics also supported the above conclusion. The method of studying the protein during Clostridium species fermentation provides a new perspective for further study on metabolic regulation. Full article

The aim of this study was to distinguish effects due to diet composition from those triggered by ruminal pH on fermentation patterns and microbial profiles in a continuous culture system (RUSITEC). The study followed a 2 × 2 factorial design, with two diets varying in the proportions of forage and concentrate and two pH levels in the culture medium. RUSITEC fermenters were used to simulate rumen fermentation and feed digestibility, fermentation end-products, microbial protein synthesis, microbial community, and long-chain fatty acid profiles in the digesta were determined. Multivariate analyses were applied to summarize the overall results. High concentrate (34% cereal grain, 32% hay) diets were more digestible (p < 0.05) than high forage (10% cereal grain, 78% hay) diets, resulting in a greater (p < 0.05) formation of most fermentation end-products and microbial protein in the rumen. However, there were no significant (p > 0.05) differences between diets in methane production. Ciliate protozoa, anaerobic fungi, some fibrolytic bacteria, hydrogenation of oleic acid, and relative proportion of conjugated linoleic acid were increased (p < 0.05) with high forage diets. A decline in rumen pH from 6.8 to 6.4 decreased (p < 0.05) feed digestibility, protein degradability, and the daily outputs of some fermentation end-products (gas, VFA, acetate, ammonia) but had no effect (p > 0.05) on the synthesis of microbial protein, and on the output of methane, propionate, butyrate or lactate. Minor changes in microbial community profile or the fatty acid relative proportions were observed within this pH range. The overall multivariate analysis revealed a clear discrimination between high-concentrate and high-forage diets, with subtler and less-defined pH effects on ruminal fermentation and microbial communities. Full article