Search Results (225)

This study investigates the efficient biogenic production of hydrogen via the thermophilic bacterium Thermotoga neapolitana, focusing on optimising process configurations to maximise yield and productivity. To determine optimal conditions, a 1 L anaerobic bioreactor with online gas analytics was designed and tested for batch, fed-batch and continuous fermentation. A maximum hydrogen production rate of 96.1 ± 1.7 Nml·L⁻¹·h⁻¹ was observed in the continuous reactor. The optimal dilution rate was 0.07 h⁻¹. Each dilution rate was kept for ≥56 h fermentation time and resulted in yields of 2.7–3.0 mol_H2·mol_glucose⁻¹. A consistently high cell viability (97%) was also observed across various dilution rates. A detailed carbon balance indicates acetate as the main by-product, closely linked to the hydrogen production pathway. Compared to fed batch and batch, the hydrogen production rate could be increased and remain constant over a longer time. In this way the continuous reactor design showed an additional method to produce hydrogen to the established ones. Fermentative hydrogen production is particularly promising when using carbohydrate containing biomass and biowaste, as it can be considered carbon dioxide neutral. Full article

The study provides valuable insights into the sustainable utilization of edible tuber peels from the high mountains of the Argentinian Puna, which constitutes promising reserves of bioactive phenolic compounds with the potential to enhance the biofunctional properties of lactic acid bacteria. Thirty-two extracts derived from peels of different varieties of tubers, such as Oxalis tuberosa Mol., Ullucus tuberosus Caldas, and Solanum tuberosum L. were incorporated into lactobacilli cultures and individually evaluated. These selectively enhance the development of the probiotic strain Lactiplantibacillus plantarum ATCC 10241 and of Lacticaseibacillus paracasei CO1-LVP105 from ovine origin, without promoting the growth of a pathogenic bacteria set (Escherichia coli O157:H12 and ATCC 35218, Salmonella enterica serovar Typhimurium ATCC 14028, and S. corvalis SF2 and S. cerro SF16), in small amounts. To determine the main phenolic group concentrated in the phytoextracts, a bio-guided study was conducted. The most significant results were obtained by O. tuberosa phytochemicals added to the culture medium at 50 µg/mL, yielding promising increases in biofilm formation (78% for Lp. plantarum and 43% for L. paracasei) and biosurfactant activity (112% for CO1-LVP105 strain). These adaptive strategies developed by bacteria possess key biotechnological significance. Furthermore, the bio-detoxification capacity of phenol and o-phenyl phenol, particularly of the novel strain CO1-LVP105, along with its mode of action and genetic identification, is described for the first time to our knowledge. In conclusion, lactobacilli strains have potential as fermentation starters and natural products, recovered from O. tuberosa peels, and added into culture media contribute to multiple bacterial biotechnological applications in both health and the environment. Full article

Mushrooms are eukaryotic organisms with absorptive heterotrophic nutrition, capable of feeding on organic matter rich in cellulose and lignocellulose. Since ancient times, they have been considered allies and, in certain cultures, they were seen as magical beings or food of the gods. Of the great variety of edible mushrooms identified worldwide, less than 2% are traded on the market. Although mushrooms have been valued for their multiple nutritional and healing benefits, some cultures perceive them as toxic and do not accept them in their culinary practices. Despite the existing skepticism, several researchers are promoting the potential of edible mushrooms. There are two main methods of mushroom cultivation: solid-state fermentation and submerged fermentation. The former is the most widely used and simplest, since the fungus grows in its natural environment; in the latter, the fungus grows suspended without developing a fruiting body. In addition, submerged fermentation is easily monitored and scalable. Both systems are important and have their limitations. This article discusses the main methods used to increase the performance of submerged fermentation with emphasis on the modes of operation used, types of bioreactors and application of morphological bioengineering of filamentous fungi, and especially the use of intelligent automatic control technologies and the use of non-invasive monitoring in fermentation systems thanks to the development of machine learning (ML), neural networks, and the use of big data, which will allow more accurate decisions to be made in the fermentation of filamentous fungi in submerged environments with improvements in production yields. Full article

One of the world’s most sustainable solutions is to replace fossil-based polymers with biopolymers. The production of xanthan gum can be optimized using various renewable and cost-effective raw materials, which is a key focus in industrial biotechnology. Xanthan gum is a bioengineered thickening, stabilizing, and emulsifying agent. It has unique properties for use in many industries (food, biotechnology, petrochemicals, agricultural, cosmetics, wastewater treatment) and medical applications. It is tasteless, environmentally safe, non-toxic, and biodegradable. The biotechnological production of xanthan gum depends on several factors: bacterial strain development, culture medium preparation, carbon sources, fermentation parameters and modes, pH, temperature, recovery, purification, and quality control regulations. Bio-innovative strategies have been developed to optimize the production of xanthan gum. A variety of carbon and nitrogen sources, as well as alternative renewable sources, have been used in the production of xanthan gum. The aim of the present study was to optimize the xanthan gum yield using Xanthomonas campestris bacteria and different carbon (D-glucose, D-sorbitol, lactose, sucrose, D-mannitol, D-fructose, erythritol, coconut palm sugar, L-arabinose, unrefined cane sugar), various nitrogen (bacterial peptone, casein peptone, L-glutamic acid, L-arginine, L-methionine, L-tryptophan, malt extract, meat extract, L-phenylalanine, soy peptone) and alternative carbon (orange peels, tangerine peels, lemon peels, avocado peels, melon peels, apple peels, cellulose, xylose, xylitol) sources. The xanthan gum samples were analyzed using antioxidant methods. Our study showed that using L-glutamic acid as the carbon source for 72 h of bacterial fermentation of Xanthomonas campestris resulted in the highest xanthan gum yield: 32.34 g/L. However, using renewable resources, we achieved a very high concentration of xanthan gum in just 24 h of fermentation. According to the reducing power and DPPH methods, the highest antioxidant activities were measured for xanthan gum whose biosynthesis was based on renewable resources. Xanthan gum structures have been verified by FT-IR and ¹H NMR analysis. The sustainable biotechnology study has the advantage of increasing the sustainable production of xanthan gum by using renewable alternative resources compared to other production processes. Xanthan gum continues to be a valuable biopolymer with a wide range of industrial applications while promoting environmentally friendly production practices. Full article

Fermentation is one of the oldest food processing techniques and is widely utilized in dairy product processing, during which nutrient availability and bioactive compounds are altered. However, the complete mode of action by which fermented dairy exerts beneficial effects on the host remains unknown. The present study investigated the effect of milk and yogurt ingestion alone or combined with prebiotic inulin on the transcriptome of colonic mucosa, liver, and femur in healthy rats. Young growing male rats were fed one of four experimental diets containing (1) skimmed milk, (2) skimmed milk supplemented with inulin (5% w/w), (3) yogurt, or (4) yogurt supplemented with inulin (5% w/w) for 6 weeks. Microarray results revealed that yogurt consumption resulted in 2195 upregulated differential expressed genes (DEGs) and 1474 downregulated DEGs in colonic mucosa as compared with milk consumption. According to Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, tight junction-, immune system-related pathways in the colonic mucosa and metabolic pathways in the liver were enriched with yogurt consumption. No evident differences were identified in the bone transcriptome between the diet groups. In conclusion, the study found that the intake of fermented dairy exerts more pronounced effects on gene expression in the intestinal tissue than prebiotics supplementation. Full article

Constipation is a common problem which often causes negative impacts on the patient’s quality of life. Apart from the pharmacologic and diet approaches, the use of probiotics has gradually shown promising efficacy to alleviate constipation. However, an exact understanding of the underlying mechanisms of probiotic actions on alleviating constipation is still unclear and need to be explored. In this review, we propose a model, ‘probiotics in human colon’, from a bioprocess engineering perspective. This model can be interpreted as a new concept of bionic colonic bioreactor design of a human colon in vitro, in which the transport phenomena during the fermentation of chyme by probiotics can be detected. By reviewing the anatomy structure and peristalsis mode of the human colon, we have focused on the influence by probiotics on the physical properties of colonic contents during the fermentation process. We relate physical properties such as shape, water content, density, hardness, viscosity, and elasticity to constipation symptoms directly. The influences on the physical properties of colon contents triggered by probiotics can be a potential key to understand the mechanisms for alleviating constipation. Full article

The essential amino acid tryptophan yields a plethora of secondary metabolites with key roles in plants and animals. Its fate in different living organisms is crucial for their own health, and metabolic profiling is a valuable tool for investigating it. Among the various metabolites, those retaining the indole structure were examined for qualitative and quantitative profiling. Liquid chromatography coupled with a tandem mass spectrometry detector with an electrospray ionization source (HPLC-ESI-MS/MS), acquiring in multiple reaction monitoring (MRM) mode, was used to develop a selective and sensitive method for the simultaneous analysis of tryptophan and 10 indole structure-retaining metabolites of it. Satisfactory values were obtained for linearity (R² ≥ 0.99 for all compounds except two), sensitivity (LOD, within 6–31 ng/mL, and LOQ, within 17–94 ng/mL, where minimum and maximum values were relative to serotonin and 5-methoxytryptamin, respectively), reproducibility (interday and intraday precision and accuracy), and effect of the matrix (recovery and matrix effect). The method was then successfully applied to the analysis of different types of beverage, such as herbal products, like Eschscholzia californica and a sleep herbal tea marketed with added melatonin (consumed to reduce anxiety and improve sleep quality), and fermented beverages, like beer and kefir. High amounts of tryptophan (from 77 ng/mL in kefir to 26,974 ng/g in the sleep herbal tea) followed by lower contents of serotonin (from 29 ng/mL in kefir to 2207 ng/g in the sleep herbal tea), were found in all samples along with the serotonin pathway-related compounds 5-hydroxytryptophan and tryptamine. Melatonin was detected in the plant matrix Eschscholzia c. for the first time to our knowledge (446 ng/g) and in the fermented beverages (96 ng/mL in beer and 39 ng/mL in kefir), regardless of their vegetable or animal origin, along with the melatonin route metabolites 5-methoxytryptamine and tryptophan ethyl ester. The amount of melatonin in the sleep herbal tea (556,464 ng/g) was in strong agreement with the declared content. Suggested applications include the search for biomarkers in phytochemical characterization, mechanistic studies of tryptophan’s chemistry, valorization of foods, beverages, and tryptophan-rich agro-food by-products and waste for nutraceutical and pharmacological purposes. Full article

For efficient production of microbial lipids also known as single cell oil (SCO), selection of favorable growth conditions including the substrate for maximum conversion into storage lipids is imperative. Utilization of lignocellulosic biomass for microbial oil production is a promising approach as it is renewable, sustainable, and available in abundance, with a significant quantity of fermentable sugars. Because of their intricate structure and biomolecular composition, lignocellulosic substrates exhibit high recalcitrance and demand specific pretreatments to release the fermentable sugars. However, pretreating the lignocellulosic substrate not only produces assimilable sugars but also various fermentation inhibitors that can significantly impede microbial growth and/or lipogenesis. Therefore, in this review, we discuss different inhibitors present in the lignocellulosic hydrolysates, and the impact on oleaginous microbial growth and metabolic activity, particularly concerning lipid production. Furthermore, the mode of inhibition of the various inhibitors and potential strategies to detoxify these are discussed in this review. Full article

The primary aim of this work was to share the results from a Research Project supported by the Italian National Research Council, which led to the development of a versatile jacketed tower bioreactor. Designed to optimize oxygen transfer efficiency and process control, the reactor incorporated a reciprocating air compressor, centrifugal pumps, a draft tube with or without perforated plates, and a series of gas–liquid ejectors. Its flexible design enabled operation in both airlift and ejector-loop modes, making it suitable for a wide range of aerobic fermentation processes. By sharing the detailed engineering design, operational procedures of this pilot-scale bioreactor, as well as its performance data when cultivating yeasts on whey and potato wastewater, a detailed blueprint was given to researchers seeking to advance bioreactor technology, particularly in the context of emerging fields like cultured meat production, pharmaceutical manufacturing, and environmental bioremediation. Full article

Microbial foods include microbial biomass, naturally fermented foods, and heterologously synthesized food ingredients derived from microbial fermentation. Terpenoids, using isoprene as the basic structure, possess various skeletons and functional groups. They exhibit diverse physicochemical properties and physiological activities, such as unique flavor, anti-bacterial, anti-oxidant, anti-cancer, and hypolipemic, making them extensively used in the food industry, such as flavor, fragrance, preservatives, dietary supplements, and medicinal health food. Compared to traditional strategies like direct extraction from natural species and chemical synthesis, microbial cell factories for edible terpenoids have higher titers and yields. They can utilize low-cost raw materials and are easily scaling-up, representing a novel green and sustainable production mode. In this review, we briefly introduce the synthetic pathway of terpenoids and the applications of microbial cell factories producing edible terpenoids. Secondly, we highlight several typical and non-typical microbial chassis in edible terpenoid-producing cell factories. In addition, we reviewed the recent advances of representative terpenoid microbial cell factories with a gram-scale titer in food flavor, food preservation, nutritional enhancers, and medicinal health foods. Finally, we predict the future directions of microbial cell factories for edible terpenoids and their commercialization process. Full article

Direct-fed microbials (DFMs) have shown the potential to improve livestock performance and overall health. Extensive research has been conducted to identify new DFMs and understand their mechanisms of action in the gut. Bacillus species are multifunctional spore-forming bacteria that exhibit resilience to harsh conditions, making them ideal candidates for applications in the feed industry and livestock production. This study investigates the mode of action of B. licheniformis and B. subtilis in the rumen using diverse in vitro techniques. Our results revealed that both strains germinated and grew in sterile rumen and intestinal contents from dairy cows and bulls. Gas composition analysis of in vitro cultures in a medium containing 40% rumen fluid demonstrated that germination of B. licheniformis and B. subtilis strains reduced oxygen levels, promoting an anaerobic environment favorable to rumen microbes. Enzymatic activity assays showed that B. licheniformis released sugars from complex substrates and purified polysaccharides in filtered rumen content. Additionally, the combination of B. licheniformis and B. subtilis survived and grew in the presence of a commercial monensin dose in rumen fluid media. The effects of B. licheniformis and B. subtilis on rumen fermentation activity and microbiota were studied using an in vitro batch fermentation assay. In fermenters that received a combination of B. licheniformis and B. subtilis, less CO₂ was produced while dry matter degradation and CH₄ production was comparable to the control condition, indicating better efficiency of dry matter utilization by the microbiota. The investigation of microbiota composition between supplemented and control fermenters showed no significant effect on alpha and beta diversity. However, the differential analysis highlighted changes in several taxa between the two conditions. Altogether, our data suggests that the administration of these strains of Bacillus could have a beneficial impact on rumen function, and consequently, on health and performance of ruminants. Full article

Lactic acid bacteria (LAB) constitute a heterogeneous group of bacteria isolated from fermented foods, animals, plants, and mammalian guts, with many health-promoting properties. Probiotics with antagonistic properties against human pathogens and foodborne bacteria have garnered significant attention from the scientific fraternity. A dedicated review focusing on plant-derived probiotic bacteria and their antagonistic properties has not been comprehensively reviewed. Thus, this review aimed at providing an overview of LAB isolates derived from several unconventional sources such as fruits, seeds, fruit pulp, leaves, roots, vegetables, grasses, and flowers and with their antibacterial, antifungal, and antiviral properties. This paper reviewed the antimicrobial properties of different genera, Lactobacillus, Leuconostoc, Weissella, Enterococcus, Pediococcus, Bacillus, and Fructobacillus, their postbiotics, and paraprobiotics. Several important mechanisms, including the secretion of bacteriocins, bacteriocin-like substances, reuterin, organic acids (lactic and acetic), peptides, exopolysaccharides, and hydrogen peroxide, have been attributed to their antimicrobial actions against pathogens. However, their precise mode of action is poorly understood; hence, further research should be conducted to reveal detailed mechanisms. Finally, the review discusses the summary and future implications. Given the significance, LAB and derived antimicrobial compounds can potentially be exploited in food preservation and safety or for medicinal applications after evaluating their safety. Full article

Stingless bees are vital pollinators in tropical and neotropical regions, emphasizing the need to conserve these species. However, resource scarcity, particularly pollen, during certain periods negatively impacts bee health and pollination efficiency. To address this, we developed a fermented protein feed using microorganisms from pollen of Melipona quadrifasciata, a species commonly found in the Brazilian Atlantic Forest. The fermented feed consisted of a protein bran mixture, sugar syrup, and an inoculant derived from species’ fermented pollen. To assess the feed quality, nutritional evaluation and metabolomic analyses were conducted (UHPLC/DAD-HRMS/MS and GC-MS). The metabolomic analysis identified 85 compounds in positive and 102 in negative ionization modes. Shared metabolites, such as isoquercetin and palatinose, highlighted similarities between the feed and natural fermented pollen. Laboratory tests with caged bees showed comparable consumption between the feed and fermented pollen with no statistical difference (mean: 0.067 g vs. 0.060 g; p = 0.53). The feed exhibited a favorable pH transition (5.30 pre-fermentation to 4.41 after the fermentation process) and remained stable for six months at ±2 °C. These findings demonstrate a biotechnological advance in stingless bee nutrition, offering a viable solution during resource scarcity and promoting the health of M. quadrifasciata. Full article

Although the mitochondrial genome is an attribute of all eukaryotes, some yeast species (called petite-positive) can replicate without mitochondrial DNA (mtDNA). Strains without mtDNA (known as rho⁻ mutants or petite mutants) are respiration-deficient and require fermentable carbon sources (such as glucose) for their metabolism. However, they are compromised in many aspects of fitness and competitiveness. Nevertheless, a few research groups have reported that some petite mutants of Candida glabrata and Saccharomyces cerevisiae manifested higher levels of tolerance to the antifungal fluconazole than their wild-type (WT) counterparts. In this study, we show that elevated tolerance to two or three out of four tested antifungals is a generic feature of at least five petite-positive species of yeasts including C. glabrata (higher tolerance of petites to clotrimazole and miconazole), S. bayanus (tolerance to clotrimazole, fluconazole, and miconazole), S. cerevisiae (tolerance to clotrimazole and fluconazole), S. paradoxus (tolerance to clotrimazole, fluconazole, and miconazole), and S. pastorianus (tolerance to clotrimazole and fluconazole). Comparing the levels of tolerance to the antifungals in WT and petite mutants was based on measuring the diameters of the zones of inhibition (ZOIs) using disc diffusion assays. The mode of inhibition in the majority of WT strains by all antifungals was fungicidal; most of the rho⁻ mutants manifested fungistatic inhibition. We observed partial (not complete) inhibition in WT, with four different types of ZOI patterns that were species- and antifungal-specific. The partial inhibition was characterised by the presence of antifungal-tolerant colonies within ZOI areas. The inability of these colonies selected from ZOIs to grow on glycerol, as a single source of carbon, proved that they were rho⁻ mutants spontaneously generated in the WT populations. The results on the elevated tolerance of petite strains to antifungals are discussed in terms of the prospective positive selection of respiratory-deficient mutants and the various implications of such selection. Full article

Protocatechuate acid (PCA) is a phenolic acid naturally synthesized by various organisms. Protocatechuic acid is synthesized by plants for physiological, metabolic functions, and self-defense, but extraction from plants is less efficient compared to the microbial culture process. The microbial synthesis of protocatechuic acid is sustainable and, due to its high yield, can save energy consumption when producing the same amount. To enhance PCA production using Corynebacterium glutamicum, a statistical optimization of the production medium was performed using full factorial design, the steepest ascent method, and the response surface method. The optimized production medium enabled a PCA production of over 5 g/L in a 72 h batch culture. However, PCA cytotoxicity affected the strain growth and PCA production rate, with an inhibitory concentration of approximately 5 g/L in the fermentation broth. Finally, continuous fermentation was operated for 150 h in the steady-state mode, maintaining the concentration of PCA below 5 g/L. The optimization method established in this study successfully increased PCA production levels, and the findings presented herein are anticipated to contribute to the industrialization of PCA production using C. glutamicum. Full article