Improvement of the freezing resistance characteristics of yeast in dough starter.

Improving the freezing resistance of yeast in dough starters is one of the most effective methods to promote the healthy development of frozen dough technology. When the dough starter was composed of yeast, lactic acid bacteria and acetic acid bacteria, the microbial proportion was 10:1:5, and the ratio of wheat flour to corn flour was 1:1. The proline contents of the starters and the survival rates and fermentation capacity of yeast significantly increased compared with those of the starter composed of yeast and wheat flour only (P < 0.05). Laser confocal microscopy observation showed that the cell membrane damage of yeast obviously decreased. Low-field nuclear magnetic resonance method revealed that the water distribution state of starters changed. Adding corn flour and acetic acid bacteria to dough starter in appropriate proportions improves yeast freezing resistance.

Metabolic Engineering of Corynebacterium glutamicum for the High-Level Production of l-Valine under Aerobic Conditions.

l-Valine, an essential amino acid, serves as a valuable compound in various industries. However, engineering strains with both high yield and purity are yet to be delivered for microbial l-valine production. We engineered a Corynebacterium glutamicum strain capable of highly efficient production of l-valine. We initially introduced an acetohydroxy acid synthase mutant from an industrial l-valine producer and optimized a cofactor-balanced pathway, followed by the activation of the nonphosphoenolpyruvate-dependent carbohydrate phosphotransferase system and the introduction of an exogenous Entner-Doudoroff pathway. Subsequently, we weakened anaplerotic pathways, and attenuated the tricarboxylic acid cycle via start codon substitution in icd, encoding isocitrate dehydrogenase. Finally, to balance bacterial growth and l-valine production, an l-valine biosensor-dependent genetic circuit was established to dynamically repress citrate synthase expression. The engineered strain Val19 produced 103 g/L of l-valine with a high yield of 0.35 g/g glucose and a productivity of 2.67 g/L/h. This represents the highest reported l-valine production in C. glutamicum via direct fermentation and exhibits potential for its industrial-scale production, leveraging the advantages of C. glutamicum over other microbes.

Supplementation of date palm (Phoenix dactylifera L.) residue for growth and lactic acid production of probiotic bacterial Lactobacillus spp.

Date palm is an age-old cultivated plant that thrives in tropical and subtropical regions. The date palm is a bountiful source of carbohydrates, encompassing sucrose, glucose, and fructose and proteins. The date industry generates a significant volume of unused by-products. Dates offer a diverse range of by-products beyond the agri-food sector. LAB have garnered extensive utilisation across diverse food sectors, spanning meat, vegetables, beverages, dairy products, and other fermented foods. In the quest for establishing a new large-scale fermentation process for lactic acid there has been a concerted effort to utilise more cost-effective medium components. In the present work, date palm residue (DPR) derived from date palm fruit, after sugar extraction, was incorporated into MRS. The fermentation process was executed through two distinct fermentation systems. Initially, experiments were conducted in flasks. Afterward, the optimal conditions for bacterial growth were determined, and the experiment was carried out using a bioreactor. DPR supported the probiotic Lactobacillus spp. growth especially after 48 h incubation. The prebiotic effect of DPR on Lactobacillus spp. was reported. An increase in the total number of bacterial populations was observed in response to the addition the DPR until 48 h. Specifically, the supplementing DPR at a concentration of 1.5% in batch fermentation enhanced the growth and lactic acid production of Lactobacillus casei. This study suggests that DPR could potentially function as an economical prebiotic source and could be seamlessly incorporated as a functional food ingredient, thereby transforming a waste product into an economically sustainable food substrate.

An efficient cre-based workflow for genomic integration and expression of large biosynthetic pathways in Eubacterium limosum.

Acetogenic Clostridia are obligate anaerobes that have emerged as promising microbes for the renewable production of biochemicals owing to their ability to efficiently metabolize sustainable single-carbon feedstocks. Additionally, Clostridia are increasingly recognized for their biosynthetic potential, with recent discoveries of diverse secondary metabolites ranging from antibiotics to pigments to modulators of the human gut microbiota. Lack of efficient methods for genomic integration and expression of large heterologous DNA constructs remains a major challenge in studying biosynthesis in Clostridia and using them for metabolic engineering applications. To overcome this problem, we harnessed chassis-independent recombinase-assisted genome engineering (CRAGE) to develop a workflow for facile integration of large gene clusters (>10 kb) into the human gut acetogen Eubacterium limosum. We then integrated a non-ribosomal peptide synthetase gene cluster from the gut anaerobe Clostridium leptum, which previously produced no detectable product in traditional heterologous hosts. Chromosomal expression in E. limosum without further optimization led to production of phevalin at 2.4 mg/L. These results further expand the molecular toolkit for a highly tractable member of the Clostridia, paving the way for sophisticated pathway engineering efforts, and highlighting the potential of E. limosum as a Clostridial chassis for exploration of anaerobic natural product biosynthesis.

A concise review of the nutritional profiles, microbial dynamics, and health impacts of fermented mushrooms.

Mushrooms have garnered significant attention for their nutritional composition and potential health benefits, including antioxidant, antihypertensive, and cholesterol-lowering properties. This review explores the nutritional composition of edible mushrooms, including their high protein content, essential amino acids, low fat, cholesterol levels, and bioactive compounds with medicinal value. Moreover, the study analyzes the microbiology of mushroom fermentation, focusing on the diverse microbial ecosystem involved in the transformation of raw mushrooms and the preservation methods employed to extend their shelf life. Special emphasis is placed on lactic acid fermentation as a cost-effective and efficient preservation technique. It involves controlling the growth of lactic acid bacteria to enhance the microbial stability and nutritional quality of mushrooms. Furthermore, the bioactivities of fermented mushrooms are elucidated, which are antioxidant, antimicrobial, anticancer, anti-glycemic, immune modulatory, and other biological activities. The mechanisms underlying these bioactivities are explored, emphasizing the role of fermented mushrooms in suppressing free radicals, enhancing antioxidant defenses, and modulating immune responses. Overall, this review provides comprehensive insights into the nutritional composition, microbiology, bioactivities, and underlying mechanisms of fermented mushrooms, highlighting their potential as functional foods with significant health-promoting properties.

Sensoproteomic Characterization of Lactobacillus Johnsonii-Fermented Pea Protein-Based Beverage: A Promising Strategy for Enhancing Umami and Kokumi Sensations while Mitigating Bitterness.

This study investigated the mechanism underlying the flavor improvement observed during fermentation of a pea protein-based beverage using Lactobacillus johnsonii NCC533. A combination of sensomics and sensoproteomics approach revealed that the fermentation process enriched or generated well-known basic taste ingredients, such as amino acids, nucleotides, organic acids, and dipeptides, besides six new taste-active peptide sequences that enhance kokumi and umami notes. The six new umami and kokumi enhancing peptides, with human recognition thresholds ranging from 0.046 to 0.555 mM, are produced through the degradation of Pisum sativum's storage protein. Our findings suggest that compounds derived from fermentation enhance umami and kokumi sensations and reduce bitterness, thus improving the overall flavor perception of pea proteins. In addition, the analysis of intraspecific variations in the proteolytic activity of L. johnsonii and the genome-peptidome correlation analysis performed in this study point at cell-wall-bound proteinases such as PrtP and PrtM as the key genes necessary to initiate the flavor improving proteolytic cascade. This study provides valuable insights into the molecular mechanisms underlying the flavor improvement of pea protein during fermentation and identifies potential future research directions. The results highlight the importance of combining fermentation and senso(proteo)mics techniques in developing tastier and more palatable plant-based protein products.

Sustainable diets with plant-based proteins require considerations for prevention of proteolytic fermentation.

The human diet requires a more plant-based approach due to the exhaustive effects animal-based foods have on the environment. However, plant-based proteins generally miss a few or have a lower variety in essential amino acids and are more difficult to digest. Subsequently they might be prone to fermentation by the microbiome in the proximal colon. Proteolytic fermentation can induce microbial-metabolites with beneficial and negative health effects. We review current insight into how balances in saccharolytic and proteolytic fermentation can be maintained when the diet consists predominantly of plant-based proteins. Some proteolytic fermentation metabolites may negatively impact balances in gut microbiota composition in the large intestine and influence immunity. However, proteolytic fermentation can potentially be prevented in the proximal colon toward more saccharolytic fermentation through the addition of non-digestible carbohydrates in the diet. Knowledge on this combination of plant-based proteins and non-digestible carbohydrates on colonic- and general health is limited. Current data suggest that transitioning toward a more plant-based protein diet should be accompanied with a consumption of increased quantities and more complex structures of carbohydrates or by application of technological strategies to enhances digestibility. This can reduce or prevent proteolytic fermentation which might consequently improve human health.

Fermentation biotechnology applied to wheat bran for the degradation of cell wall fiber and its potential health benefits: A review.

Consumption of wheat bran is associated with health benefits. However, the insoluble cell layer fiber and considerable levels of anti-nutritional factors limit bioavailability of wheat bran, which can be effectively improved through fermentation. To comprehensively elucidate the precise biotransformation and health benefits mechanisms underlying wheat bran fermentation. This review investigates current fermentation biotechnology for wheat bran, nutritional effects of fermented wheat bran, mechanisms by which fermented wheat bran induces health benefits, and the application of fermented wheat bran in food systems. The potential strategies to improve fermented wheat bran and existing limitations on its application are also covered. Current findings support that microorganisms produce enzymes that degrade the cell wall fiber of wheat bran during the fermentation, releasing nutrients and producing new active substances while degrading anti-nutrient factors in order to effectively improve nutrient bioavailability, enhance antioxidant activity, and regulate gut microbes for health effects. Fermentation has been an effective way to degrade cell wall fiber, thereby improving nutrition and quality of whole grain or bran-rich food products. Currently, there is a lack of standardization in fermentation and human intervention studies. In conclusion, understanding effects of fermentation on wheat bran should guide the development and application of bran-rich products.

Efficient synthesis of limonene production in Yarrowia lipolytica by combinatorial engineering strategies.

BACKGROUND: Limonene has a variety of applications in the foods, cosmetics, pharmaceuticals, biomaterials, and biofuels industries. In order to meet the growing demand for sustainable production of limonene at industry scale, it is essential to find an alternative production system to traditional plant extraction. A promising and eco-friendly alternative is the use of microbes as cell factories for the synthesis of limonene. RESULTS: In this study, the oleaginous yeast Yarrowia lipolytica has been engineered to produce D- and L-limonene. Four target genes, l- or d-LS (limonene synthase), HMG (HMG-CoA reductase), ERG20 (geranyl diphosphate synthase), and NDPS1 (neryl diphosphate) were expressed individually or fused together to find the optimal combination for higher limonene production. The strain expressing HMGR and the fusion protein ERG20-LS was the best limonene producer and, therefore, selected for further improvement. By increasing the expression of target genes and optimizing initial OD, 29.4 mg/L of L-limonene and 24.8 mg/L of D-limonene were obtained. We also studied whether peroxisomal compartmentalization of the synthesis pathway was beneficial for limonene production. The introduction of D-LS and ERG20 within the peroxisome improved limonene titers over cytosolic expression. Then, the entire MVA pathway was targeted to the peroxisome to improve precursor supply, which increased D-limonene production to 47.8 mg/L. Finally, through the optimization of fermentation conditions, D-limonene production titer reached 69.3 mg/L. CONCLUSIONS: In this work, Y. lipolytica was successfully engineered to produce limonene. Our results showed that higher production of limonene was achieved when the synthesis pathway was targeted to the peroxisome, which indicates that this organelle can favor the bioproduction of terpenes in yeasts. This study opens new avenues for the efficient synthesis of valuable monoterpenes in Y. lipolytica.

Effects of probiotic-supplemented milk replacer on growth, blood biochemistry, fermentation, digestibility, and carcass traits in lambs.

The objective of this study was to investigate the effects of feeding cow's milk replacer and probiotic supplementation on growth performance, blood metabolites, ruminal fermentation parameters, and nutrient digestibility in male lambs, with ewe's milk serving as the control treatment. Eighteen male lambs with an average initial body weight of 5.0 ± 1.2 kg and age of 15 ± 4 days were randomly assigned to three experimental groups: ewe's milk, cow's milk replacer, and cow's milk replacer with probiotic supplementation. The experiment was conducted using a completely randomized design. Lambs fed ewe's milk showed significantly higher average daily gain (218.4 g/day) compared to those fed cow's milk replacer (183.7 g/day) or cow's milk replacer with probiotic (209.1 g/day). Similarly, dry matter intake was highest in the ewe's milk group (585.6 g/day) compared to the cow's milk replacer (435.9 g/day) and cow's milk replacer with probiotic (510.5 g/day) groups. Blood glucose levels were higher in the ewe's milk group (75.3 mg/dL) compared to the cow's milk replacer (70.3 mg/dL) and cow's milk replacer with probiotic (72.1 mg/dL) groups. Probiotic supplementation resulted in increased blood urea nitrogen (15.6 mg/dL) and total protein (7.3 g/dL) levels compared to the other groups. Furthermore, the ewe's milk group showed higher apparent dry matter (76.1%) and crude protein (68.5%) digestibility compared to the other treatments. The fecal score on day 30 was higher in the cow's milk replacer group (2.34) compared to the ewe's milk (1.24) and cow's milk replacer with probiotic (1.45) groups. There were no significant differences in the carcass traits of the experimental lambs. In conclusion, based on the results of this study, it seems that feeding cow's milk + probiotic as a replacement for ewe's milk did not significantly affect the performance, carcass traits, and blood and fermentation parameters in infant lambs.

Process optimization of pearl millet-based fermented beverage components using response surface methodology and evaluation of its physicochemical parameters.

The majority of millets are produced in India, particularly pearl millet, which is more nutritious than both wheat and rice. Native to India, the "north-western semi-arid region" produces rabadi, a natural dairy beverage made from cereal and fermented by lactic acid bacteria. The three components of rabadi viz. pearl millet, buttermilk and deionized water were optimized by using Design Expert software trial version13.0.12.0. Rabadi was processed by using the traditional technique i.e., the three components were mixed in sterile conditions and fermented for 4 h at 37 °C and then cooked for 7-8 min at high flame and finally boiled. Parameters such as pH, viscosity, ash, moisture, total solids, antioxidants, total phenols, tannins, suspension stability, titratable acidity, total sugars, and reducing sugars were analysed for all 16 runs predicted by the software. 6.83 g of pearl millet, 42.44 ml of buttermilk, and 50.72 ml of deionized water were predicted to be the best formulation of rabadi, when using the set goal of maximizing the antioxidants, total phenols and minimizing the tannins. FTIR analysis was also carried out, after the final concentration optimization, to confirm the presence of phenolic compounds, antioxidants, carbohydrates, proteins and fatty acids.

The flavour of wheat gluten hydrolysate after Corynebacterium Glutamicum fermentation: Effect of degrees of hydrolysis and fermentation time.

Corynebacterium glutamicum was used to ferment wheat gluten hydrolysates (WGHs) to prepare flavour base. This study investigated the effect of hydrolysis degrees (DHs) and fermentation time on flavour of WGHs. During fermentation, the contents of amino nitrogen, total acid and small peptides increased, while the protein and pH value decreased. Succinic acid, GMP, and Glu were the prominent umami substances in fermented WGHs. The aromas of WGHs with different DHs could be distinguished by electronic nose and GC-IMS. Based on OAV of GC-MS, hexanal was the main compound in WGHs, while phenylethyl alcohol and acetoin were dominant after fermentation. WGHs with high DHs accumulated more flavour metabolites. Correlation analysis showed that small peptides (<1 kDa) could promote the formation of flavour substances, and Asp was potentially relevant flavour precursor. This study indicated that fermented WGHs with different DHs can potentially be used in different food applications based on flavour profiles.

Genomic Modifications of Lactic Acid Bacteria and Their Applications in Dairy Fermentation.

Lactic Acid Bacteria (LAB) have a long history of safe use in milk fermentation and are generally recognized as health-promoting microorganisms when present in fermented foods. LAB are also important components of the human intestinal microbiota and are widely used as probiotics. Considering their safe and health-beneficial properties, LAB are considered appropriate vehicles that can be genetically modified for food, industrial and pharmaceutical applications. Here, this review describes (1) the potential opportunities for application of genetically modified LAB strains in dairy fermentation and (2) the various genomic modification tools for LAB strains, such as random mutagenesis, adaptive laboratory evolution, conjugation, homologous recombination, recombineering, and CRISPR (clustered regularly interspaced short palindromic repeat)- Cas (CRISPR-associated protein) based genome engineering. Lastly, this review also discusses the potential future developments of these genomic modification technologies and their applications in dairy fermentations.

Engineering Escherichia coli for efficient glutathione production.

Glutathione is a tripeptide of excellent value in the pharmaceutical, food, and cosmetic industries that is currently produced during yeast fermentation. In this case, glutathione accumulates intracellularly, which hinders high production. Here, we engineered Escherichia coli for the efficient production of glutathione. A total of 4.3 g/L glutathione was produced by overexpressing gshA and gshB, which encode cysteine glutamate ligase and glutathione synthetase, respectively, and most of the glutathione was excreted into the culture medium. Further improvements were achieved by inhibiting degradation (Δggt and ΔpepT); deleting gor (Δgor), which encodes glutathione oxide reductase; attenuating glutathione uptake (ΔyliABCD); and enhancing cysteine production (PompF-cysE). The engineered strain KG06 produced 19.6 g/L glutathione after 48 h of fed-batch fermentation with continuous addition of ammonium sulfate as the sulfur source. We also found that continuous feeding of glycine had a crucial role for effective glutathione production. The results of metabolic flux and metabolomic analyses suggested that the conversion of O-acetylserine to cysteine is the rate-limiting step in glutathione production by KG06. The use of sodium thiosulfate largely overcame this limitation, increasing the glutathione titer to 22.0 g/L, which is, to our knowledge, the highest titer reported to date in the literature. This study is the first report of glutathione fermentation without adding cysteine in E. coli. Our findings provide a great potential of E. coli fermentation process for the industrial production of glutathione.

Flavor assessment of a lactic fermented vinegar described in Japanese books from the Edo period (1603-1867).

Aims: Rice vinegar is a traditional fermented seasoning in Japan, and its production remained unchanged for over 800 years until the Edo period. However, based on the available information regarding rice vinegar production methods from this period and the results of reproduction experiments, we speculated that unlike the modern-day acetic fermented vinegar, rice vinegar produced during the Edo period was lactic fermented. Main methods: To verify this assumption, we analyzed the flavor components of Honcho, a lactic fermented product prepared using a method described in books, including "Honchoshokkan" from the Edo period, by capillary electrophoresis/time-of-flight mass spectrometry, high-performance liquid chromatography, gas chromatography mass spectrometry, and taste sensor analysis. Sensory evaluation was also conducted to assess validation as a seasoning. Results: Honcho contains 2 % lactic acid, which gives it its acidity, and small amounts of other nonvolatile acids, but significantly lower levels of acetic acid (0.188 ± 0.015 g/100 mL, p < 0.01). It contains more than double the free amino acids of Kurozu, a modern rice vinegar, and more glutamic acid. Boiling to remove ethanol from yeast fermentation concentrated the free amino acids 1.5 times. Sensor taste analysis showed Honcho had weaker acidity but stronger umami taste than commercial rice vinegar. The volatile compounds related to acetic acid fermentation were significantly different between Honcho and Kurozu. Boiling increased Honcho's acidity, mainly through non-volatile acids. Significance: These findings provide evidence to indicate that Honcho was an acidic seasoning for heat-cooking, which is uncommon in Japanese cuisine today and is mentioned in Edo period books. This seasoning contains many amino acids, implying that it adds umami flavor, not only the sourness of modern vinegar.

Rosa roxburghii tratt residue as an alternative feed for improving growth, blood metabolites, rumen fermentation, and slaughter performance in Hu sheep.

The utilization of agro-industrial by-products, such as fruit residues, presents a promising strategy for providing alternative feed to ruminants amidst rising prices and limited availability of traditional roughage. In this study, we investigated the effects of Rosa roxburghii tratt residue, a local fruit residue in Guizhou province of China, on the growth, blood metabolites, rumen fermentation, and slaughter performance of Hu sheep. Ninety-six sheep were randomly divided into four groups, namely control, treatment 1, treatment 2, and treatment 3, and fed diets containing 0, 10, 20, and 30% Rosa roxburghii Tratt residue, respectively. Feeding varying levels of Rosa roxburghii Tratt residue showed no significant differences in dry matter intake, average daily gain, or the ratio of dry matter intake to average daily gain. However, sheep in the group fed with 30% Rosa roxburghii Tratt residue showed the highest gross profit. Plasma albumin content was lower in groups fed with Rosa roxburghii Tratt residue-containing diets compared to the control group (p < 0.05). Additionally, diet treatment 3 decreased plasma creatinine levels compared to control and treatment 1 (p < 0.05). Sheep in treatment 2 and treatment 3 exhibited higher plasma high-density lipoprotein level than control and treatment 1 (p < 0.05), as well as increased total cholesterol levels compared to control (p < 0.05). There were no significant differences in other plasma metabolites. Rumen pH, N-NH3, volatile fatty acids, and methane levels did not differ significantly among the four groups. However, feeding diets treatment 2 and treatment 3 resulted in decreased water holding capacity and increased shear force compared to control and treatment 1 (p < 0.05). Furthermore, pH, red chromaticity (a*), yellowness index (b*), and luminance (L*) were unaffected among the four groups of sheep. In conclusion, the inclusion of up to 30% Rosa roxburghii Tratt residue had no adverse effects on growth performance, allowing for feed cost savings without impacting rumen fermentation parameters. Rosa roxburghii tratt residue also showed benefits in improving plasma protein efficiency and enhancing lipid metabolism, albeit with limited effects on meat quality. Considering its affordability, Rosa roxburghii Tratt residue presents a practical choice for low-cost diets, ensuring economic returns.

Sargassum horneri extract fermented by Lactiplantibacillus pentosus SH803 mediates adipocyte metabolism in 3T3-L1 preadipocytes by regulating oxidative damage and inflammation.

Sargassum horneri (S. horneri), a brown seaweed excessively proliferating along Asian coastlines, are damaging marine ecosystems. Thus, this study aimed to enhance nutritional value of S. horneri through lactic acid bacteria fermentation to increase S. horneri utilization as a functional food supplement, and consequently resolve coastal S. horneri accumulation. S. horneri supplemented fermentation was most effective with Lactiplantibacillus pentosus SH803, thus this product (F-SHWE) was used for further in vitro studies. F-SHWE normalized expressions of oxidative stress related genes NF-κB, p53, BAX, cytochrome C, caspase 9, and caspase 3, while non-fermented S. horneri (SHWE) did not, in a H2O2-induced HT-29 cell model. Moreover, in an LPS-induced HT-29 cell model, F-SHWE repaired expressions of inflammation marker genes ZO1, IL1ß, IFNγ more effectively than SHWE. For further functional assessment, F-SHWE was also treated in 3T3-L1 adipocytes. As a result, F-SHWE decreased lipid accumulation, along with gene expression of adipogenesis markers PPARγ, C/EBPα, C/EBPß, aP2, and Lpl; lipogenesis markers Lep, Akt, SREBP1, Acc, Fas; inflammation markers IFN-γ and NF-κB. Notably, gene expression of C/EBPß, IFN-γ and NF-κB were suppressed only by F-SHWE, suggesting the enhancing effect of fermentation on obesity-related properties. Compositional analysis attributed the protective effects of F-SHWE to acetate, an organic acid significantly higher in F-SHWE than SHWE. Therefore, F-SHWE is a novel potential anti-obesity agent, providing a strategy to reduce excess S. horneri populations along marine ecosystems.

Modulating metabolism through synthetic biology: Opportunities for two-stage fermentation.

Bio-based production of fuels, chemicals and materials is needed to replace current fossil fuel based production. However, bio-based production processes are very costly, so the process needs to be as efficient as possible. Developments in synthetic biology tools has made it possible to dynamically modulate cellular metabolism during a fermentation. This can be used towards two-stage fermentations, where the process is separated into a growth and a production phase, leading to more efficient feedstock utilization and thus potentially lower costs. This article reviews the current status and some recent results in application of synthetic biology tools towards two-stage fermentations, and compares this approach to pre-existing ones, such as nutrient limitation and addition of toxins/inhibitors.

Recent advancements in wastewater treatment via anaerobic fermentation process: A systematic review.

This manuscript delves into the realm of wastewater treatment, with a particular emphasis on anaerobic fermentation processes, especially dark, photo, and dark-photo fermentation processes, which have not been covered and overviewed previously in the literature regarding the treatment of wastewater. Moreover, the study conducts a bibliometric analysis for the first time to elucidate the research landscape of anaerobic fermentation utilization in wastewater purification. Furthermore, microorganisms, ranging from microalgae to bacteria and fungi, emphasizing the integration of these agents for enhanced efficiency, are all discussed and compared. Various bioreactors, such as dark and photo fermentation bioreactors, including tubular photo bioreactors, are scrutinized for their design and operational intricacies. The results illustrated that using clostridium pasteurianum CH4 and Rhodopseudomonas palustris WP3-5 in a combined dark-photo fermentation process can treat wastewater to a pH of nearly 7 with over 90% COD removal. Also, integrating Chlorella sp and Activated sludge can potentially treat synthetic wastewater to COD, P, and N percentage removal rates of 99%,86%, and 79%, respectively. Finally, the paper extends to discuss the limitations and future prospects of dark-photo fermentation processes, offering insights into the road ahead for researchers and scientists.

Mass production and characterization of an endoglucanase from Coleoptera insect (Monochamus saltuarius) in yeast Kluyveromyces lactis.

To harness the diverse industrial applications of cellulase, including its use in the food, pulp, textile, agriculture, and biofuel sectors, this study focused on the high-yield production of a bioactive insect-derived endoglucanase, Monochamus saltuarius GH Family 5 (MsGHF5). MsGHF5 was introduced into the genome of Kluyveromyces lactis to maintain expression stability, and mass production of the enzyme was induced using fed-batch fermentation. After 40 h of cultivation, recombinant MsGHF5 was successfully produced in the culture broth, with a yield of 29,000 U/L, upon galactose induction. The optimal conditions for the activity of purified MsGHF5 were determined to be a pH of 5 and a temperature of 35 °C, with the presence of ferrous ions enhancing the enzymatic activity by up to 1.5-fold. Notably, the activity of MsGHF5 produced in K. lactis was significantly higher than that produced in Escherichia coli, suggesting that glycosylation is crucial for the functional performance of the enzyme. This study highlights the potential use of K. lactis as a host for the production of bioactive MsGHF5, thus paving the way for its application in various industrial sectors.