Lactic Acid Production Research Articles

Genomic Insights Into Enterococcus mundtii 203: A Promising Probiotic Candidate Isolated From Camel Feces.

Enterococcus, a common commensal organism in the human gut, exhibits a dual nature with certain strains offering probiotic benefits, while others are associated with nosocomial infections. In this study, we conducted a comprehensive examination of the genome of Enterococcus mundtii strain 203 to assess its probiotic potential and safety profile. The complete genome sequencing, assembly, and annotation were performed, followed by bioinformatics analysis. Our investigation reveals a detailed characterization of the Enterococcus mundtii 203 genome, originally isolated from camel feces, with a size of 3,053,234 bases and a GC content of 38.4%. Importantly, our analysis suggests that this strain poses no risk as a human pathogen due to the absence of antibiotic resistance determinants and virulence factors. The genome harbors a multitude of genes responsible for lactic acid production, bioactive peptide synthesis, adhesion molecule expression, resistance to harsh gut conditions, and enhancement of host metabolism. These findings underline the potential probiotic functionality of Enterococcus mundtii 203, positioning it as a promising candidate. Notably, our study did not identify any sequences related to insertion elements or CRISPR-Cas fragments.

Association between organic nitrogen substrates and the optical purity of d-lactic acid during the fermentation by Sporolactobacillus terrae SBT-1

The development of biotechnological lactic acid production has attracted attention to the potential production of an optically pure isomer of lactic acid, although the relationship between fermentation and the biosynthesis of highly optically pure d-lactic acid remains poorly understood. Sporolactobacillus terrae SBT-1 is an excellent d-lactic acid producer that depends on cultivation conditions. Herein, three enzymes responsible for synthesizing optically pure d-lactic acid, including d-lactate dehydrogenase (D-LDH; encoded by ldhDs), l-lactate dehydrogenase (L-LDH; encoded by ldhLs), and lactate racemase (Lar; encoded by larA), were quantified under different organic nitrogen sources and concentration to study the relationship between fermentation conditions and synthesis pathway of optically pure lactic acid. Different organic nitrogen sources and concentrations significantly affected the quantity and quality of d-lactic acid produced by strain SBT-1 as well as the synthetic optically pure lactic acid pathway. Yeast extract is a preferred organic nitrogen source for achieving high catalytic efficiency of d-lactate dehydrogenase and increasing the transcription level of ldhA2, indicating that this enzyme plays a major role in d-lactic acid formation in S. terrae SBT-1. Furthermore, lactate racemization activity could be regulated by the presence of d-lactic acid. The results of this study suggest that specific nutrient requirements are necessary to achieve a stable and highly productive fermentation process for the d-lactic acid of an individual strain.

Enhanced upgrading of lignocellulosic substrates by coculture of Saccharomyces cerevisiae and Acinetobacter baylyi ADP1

BackgroundLignocellulosic biomass as feedstock has a huge potential for biochemical production. Still, efficient utilization of hydrolysates derived from lignocellulose is challenged by their complex and heterogeneous composition and the presence of inhibitory compounds, such as furan aldehydes. Using microbial consortia where two specialized microbes complement each other could serve as a potential approach to improve the efficiency of lignocellulosic biomass upgrading.ResultsThis study describes the simultaneous inhibitor detoxification and production of lactic acid and wax esters from a synthetic lignocellulosic hydrolysate by a defined coculture of engineered Saccharomyces cerevisiae and Acinetobacter baylyi ADP1. A. baylyi ADP1 showed efficient bioconversion of furan aldehydes present in the hydrolysate, namely furfural and 5-hydroxymethylfurfural, and did not compete for substrates with S. cerevisiae, highlighting its potential as a coculture partner. Furthermore, the remaining carbon sources and byproducts of S. cerevisiae were directed to wax ester production by A. baylyi ADP1. The lactic acid productivity of S. cerevisiae was improved approximately 1.5-fold (to 0.41 ± 0.08 g/L/h) in the coculture with A. baylyi ADP1, compared to a monoculture of S. cerevisiae.ConclusionThe coculture of yeast and bacterium was shown to improve the consumption of lignocellulosic substrates and the productivity of lactic acid from a synthetic lignocellulosic hydrolysate. The high detoxification capacity and the ability to produce high-value products by A. baylyi ADP1 demonstrates the strain to be a potential candidate for coculture to increase production efficiency and economics of S. cerevisiae fermentations.

Endoplasmic reticulum stress is attenuated by glycolysis in lymphatic malformations.

This study aims to investigate the role of endoplasmic reticulum stress (ER stress) in human dermal lymphatic endothelial cells (HDLECs) and lymphatic malformations (LMs) and its relationship with aerobic glycolysis and inflammation. The proliferation and apoptosis of HDLECs were examined with lipopolysaccharide (LPS) treatment. ER stress-associated proteins and glycolysis-related markers were detected by western blot. Glycolysis indexes were detected by seahorse analysis and lactic acid production assay kits. Immunohistochemistry was used to reveal the ER stress state of lymphatic endothelial cells (LECs) in LMs. LPS induced ER stress in HDLECs but did not trigger detectable apoptosis. Intriguingly, LPS-treated HDLECs also showed increased glycolysis flux. Knockdown of Hexokinase 2, a key enzyme for aerobic glycolysis, significantly inhibited the ability of HDLECs to resist ER stress-induced apoptosis. Moreover, compared to normal skin, glucose-regulated protein 78 (GRP78/BIP), and phosphorylation protein kinase R-like kinase (p-PERK), two key ER stress-associated markers, were upregulated in LECs of LMs, which was correlated with the inflected state. In addition, excessively activated ER stress inhibited the progression of LMs in rat models. These data indicate that glycolysis could rescue activated ER stress in HDLECs, which is required for the accelerated development of LMs. Inflammation enhances both ER stress and glycolysis in LECs while glycolysis is required to attenuate the pro-apoptotic effect of ER stress. Endoplasmic reticulum (ER) stress is activated in lymphatic endothelial cells (LECs) of LMs, especially in inflammatory condition. The expression of ER stress-related proteins is increased in LMs and correlated with Hexokinase 2 expression. Pharmacological activation of ER stress suppresses the formation of LM lesions in the rat model. ER stress may be a promising and effective therapeutic target for the treatment of LMs.

Kinetic analysis of lactic acid fermentation by using an extended logistic model

This study aimed to analyze the dynamics of biomass growth, lactic acid production, and glucose consumption for three microorganisms; Weissella cibaria IBUN 090-03684, Lactobacillus pantarum IBUN 090-03774, and Lactobacillus pantarum IBUN 04139 by using an extended logistic model. The microorganisms were cultivated in a 1 L bioreactor with pH control by using MRS medium at 37 ∘C for 24 h at 100 rpm. Weissella cibaria IBUN 090-03684 exhibited the highest product yield (1.26 gg−1), followed by Lactobacillus plantarum IBUN 04139 (0.96 gg−1), and Lactobacillus plantarum IBUN 090-03774 (0.90 gg−1), respectively. The results indicate that Weissella cibaria IBUN 090-03684 holds significant potential for large-scale lactic acid production. Furthermore, the reported modeling strategy fits well with experimental data for the three microorganisms evaluated.

Structural characterization and prebiotic activity of rhamnogalacturonan-I rich pumpkin pectic polysaccharide extracted by alkaline solution

The present study aimed to investigate the structural and physicochemical characteristics of alkali-extracted pectic polysaccharide (AkPP) and to evaluate its prebiotic effects. AkPP was obtained from pumpkin pulp using an alkaline extraction method. AkPP, which had a molecular weight (Mw) of mainly 13.67 kDa and an esterification degree of 9.60 %, was composed mainly of galacturonic acid (GalA), rhamnose (Rha), galactose, and arabinose. The ratio of the homogalacturonan (HG) region to the rhamnogalacturonan-I (RG-I) region in AkPP was 48.74:43.62. In the nuclear magnetic resonance spectrum, the signals indicating α-1,4-linked D-GalA, α-1,2-linked L-Rha, α-1,2,4-linked L-Rha residues were well resolved, demonstrating the presence of the HG and RG-I regions in its molecular structure. Collectively, AkPP was low methoxyl pectin rich in the RG-I region with short side chains and had a low Mw. Thermal analysis revealed that AkPP had good thermal stability. Compared to inulin, AkPP more effectively promoted the proliferation of Lactobacillus acidophilus, Lacticaseibacillus rhamnosus GG, Lacticaseibacillus casei, and Lacticaseibacillus paracasei and the production of lactic, acetic, and propionic acids. This study presents the unique structural features of AkPP and provides a scientific basis for further investigation of the potential of AkPP as a promising prebiotic.

Fusobacterium periodonticum BCT protein targeting glucose metabolism to promote the epithelial-mesenchymal transition of esophageal cancer cells by lactic acid

BackgroundThe cancer microbiota was considered the main risk factor for cancer progression. We had proved that Fusobacterium periodonticum (F.p) was higher abundance in Esophageal cancer(EC)tissues. Bioinformation analysis found that BCT was a key virulence protein of F.p. However, little is known about the role and mechanism of BCT in EC. This study aimed to recognize the key virulence protein of F.p and explore the mechanism of BCT in promoting EC.MethodsWe constructed a eukaryotic expression vector and purified the recombinant protein BCT. CCK8 used to analyzed the activity of EC after treated by different concentration of BCT. UPLC-MS/MS and ELISA used to detect the metabonomics and metabolites. The ability of migration and invasion was completed by transwell assay. RT-QPCR, WB used to analyze the expression of relevant genes.ResultsOur data showed that BCT was higher expression in EC tumor tissues (p < 0.05) and BCT in 20 µg/mL promoted the survival, invasion and migration of EC cells (EC109) (p < 0.05). Meanwhile, UPLC-MS/MS results suggested that BCT resulted in an augmentation of hypotaurine metabolism, arachidonic acid metabolism, glycolysis/gluconeogenesis, tryptophan metabolism, citrate cycle activity in EC109. The metabolic changes resulted in decreasing in glucose and pyruvate levels but increase in lactate dehydrogenase (LDH) activity and lactic acid (LA) as well as the expression of glucose transporter 1, Hexokinase 2, LDH which regulated the glycolysis were all changed (p < 0.05). The BCT treatment upregulated the expression of TLR4, Akt, HIF-1α (p < 0.05) which regulated the production of LA. Furthermore, LA stimulation promoted the expression of GPR81, Wnt, and β-catenin (p < 0.05), thereby inducing EMT and metastasis in EC109 cells.ConclusionAltogether, these findings identified that impact of BCT in regulation of glycolysis in EC109 and its involves the TLR4/Akt/HIF-1α pathway. Meanwhile, glycolysis increasing the release of LA and promote the EMT of EC109 by GPR81/Wnt/β-catenin signaling pathway. In summary, our findings underscore the potential of targeting BCT as an innovative strategy to mitigate the development of EC.

Comparative Selective Conversion of Biomass-Derived Mono- and Polysaccharides into Lactic Acid with Lanthanide Lewis Acid Catalysts

Comparative Selective Conversion of Biomass-Derived Mono- and Polysaccharides into Lactic Acid with Lanthanide Lewis Acid Catalysts

Nutritional and Molecular Studies of Alkaliphilic Tolerant Potential Amylase Producing Lactic Acid Bacteria Isolated from Indigenous Foods and Dumpsites

Alkaliphilic organisms are important in industries but most of these organisms are obtained from Bacillus species and the like that  require special scrutiny before use in fermentation of food related products. Lactic acid bacteria are generally regarded as safe (GRAS)  and are therefore readily applied in industrial fermentation with confidence. Due to high demand for the products of alkaliphilic organisms and industrial acceptability of lactic acid bacteria, there is a need to source for GRAS microorganisms that can withstand pH  ranges. The aim of this study was to isolate alkaliphilic tolerant lactic acid bacteria from fermented cassava mash, `ogi` and dump site  samples with a view to determining their nutritional requirement as well as molecular identity. Lactic acid bacteria were isolated from the  samples using standard methods. The isolates were subsequently characterized morphologically, biochemically and molecularly. Effects  of different carbon, nitrogen sources, initial temperature and pH as well as metal ions (Na+, Ca2+, Mg2+, Zn2+ and Cu2+) on the growth of  the isolates were examined. Data obtained were analyzed using one-way analysis of variance (ANOVA). Results obtained indicate that  extracted DNA sequence of A3, A7, A8 and A10 were related to those of Lactobacillus brevis FJ476121.1, Pediococcus acidilactici  CP096031.1, Pediococcus pentasaceus AB362605.1 and Pediococcus acidilactici CP053421.1 respectively with the sequences within the GenBank. 1% fructose elicited growth for all the bacteria detected except Lactobacillus brevis and ammonium nitrate was most preferred  by all the bacteria detected apart from Pediococcus pentasaceus strain. Lactobacillus brevis FJ476121.1 and Pediococcus acidilactici  CP053421.1 were the most alkaliphilic tolerant bacteria detected with OD of 0.285±0.04 and 0.198±0.04 respectively. Different metal ions  significantly influenced the growth of isolates in this study. The results in this study indicate potential applicability of the detected  bacteria for industrial processes.    

Effects of Lactobacillus plantarum (L) and molasses (M) on nutrient composition, aerobic stability, and microflora of alfalfa silage in sandy grasslands.

The objective of this experiment was to investigate the effects of Lactobacillus plantarum and molasses on the nutrient composition, fermentation quality, bacterial count, aerobic stability, and microflora of alfalfa silage in sandy grasslands. The experimental treatments included control (CK), 106 CFU/g Lactobacillus plantarum (L), 5% molasses (M), and 106 CFU/g Lactobacillus plantarum + 5% molasses (LM). The nutrient composition, fermentation quality, bacterial count, aerobic stability, and microflora were determined after 14 days and 56 days of ensiling, respectively. The results showed that the addition of L, M, and LM reduced dry matter loss (DM), neutral detergent fiber (NDF), and acid detergent fiber (ADF) content, and increased water-soluble carbohydrates (WSC) and ether extract (EE) content, compared to the CK group. Meanwhile, more lactic acid (LA) and accelerated fermentation were observed, causing the pH value to drop below 4.5 in the L, M, and LM groups after 56 days of ensiling. The addition of L, M, and LM promoted lactic acid bacteria (LAB), and inhibited yeast. The addition of L significantly increased the content of acetic acid (AA). In terms of microflora, the addition of L, M, and LM made Firmicutes become the dominant bacterial phylum earlier, while Lactobacillus, Weissella, and Pediococcus had a higher abundance. According to the result of Pearson's correlation, there is a very significant negative correlation between pH value and Lactobacillus (P < 0.01) and a very significant positive correlation between pH value and Lactococcus, Enterobacter, Enterococcus, and Leuconostoc (P < 0.01), which may be inhibited by Lactobacillus under the decreased pH value. The results of the prediction of microbial genes indicated that the addition of M could enhance the carbohydrate metabolism and membrane transport metabolism, which may contribute to LA production by LAB metabolism. In general, L, M and LM all improved the fermentation quality and reduced the loss of nutrients to varying degrees, but considering the fermentation quality, the overall effects of M and LM were better than L. M and LM are recommended to be used as silage additives in the process of alfalfa silage in sandy grasslands to improve the quality.

Evaluation of Streptococcus mutans biofilm formation and acidogenicity of infant milk formulas for treating cow milk allergy: An in vitro study.

When infants cannot consume breast milk, the most commonly available alternative milk formula is cow milk-based. Due to a rise in the prevalence of cow milk protein allergy (CMPA) among children, this study aimed to assess the biofilm formation and acidogenicity of cow milk-based formulas as well as milk formulas suggested for children with CMPA. Cow milk-based formulas with 0%, 10%, or 18% sucrose added, partially hydrolyzed formula (pHF), extensively hydrolyzed formula (eHF), amino acid-based formula (AAF), and soy-based formulas with 0%, or 11% sucrose added were evaluated. Streptococcus mutans was used as a representative microorganism associated with caries. The acidogenicity after 24-h incubation was assessed by the pH of the formed biofilm and lactic acid formation. Biofilm formation was quantified using crystal violet staining. Additionally, the biofilm characteristics were determined using confocal laser scanning microscopy (CLSM). Comparisons were made among formulas without added sucrose to observe protein-based differences. Furthermore, formulas with different sucrose percentages were compared to explore the impact of sucrose content. When comparing the formulas without added sucrose, the biofilm formation in the cow milk-based formula and pHF were significantly greater than the soy-based formula, eHF, and AAF. In the presence of S. mutans, all formulas reduced the biofilm pH below the critical enamel pH. The cow milk-based formula and AAF showed a significantly lower biofilm pH than the pHF, soy-based, and eHF groups, while the lactic acid production was markedly higher in the cow milk-based formula, pHF and AAF, compared with the eHF and soy-based formula. Adding sucrose into the cow milk-based and soy-based formulas substantially increased biofilm mass. The biofilm pH of the cow milk-based formulas, with or without sucrose, was significantly lower than that of the soy-based formulas. The CLSM indicated distinct biofilm characteristics among the different protein-based formulas, with sucrose supplementation promoting S. mutans aggregation in cow milk-based formula biofilm and increased density and intact biofilm in the soy-based formula. All assessed milk formulas had caries-inducing factors, including those without supplemental sucrose. Among them, the eHF demonstrated the least caries-inducing factors, attributed to its minimal biofilm formation and the highest biofilm pH.

Prebiotic effects of extracts from pineapple and Curcuma xanthorrhiza rhizomes towards the growth of Lacticaseibacillus paracasei

For the past few decades, plant derivatives have much been patronized in multiethnic Malaysia as traditional and complimentary medicines. Plant extracts and their phytochemical compounds have shown remarkable positive impacts on probiotics in gut. Therefore, this study was conducted to investigate the effects of several plant extracts and their phytochemical compounds on the growth of the probiotic Lacticaseibacillus paracasei (L. paracasei). Pineapple juice, pineapple waste extract, Curcuma xanthorrhiza extract, mixture of pineapple juice and Curcuma xanthorrhiza extract, inulin and mixture of inulin and bromelain were supplemented to liquid media and tested on L. paracasei at the shake flask scale for cell density, pH, postbiotic metabolites and bioactivities. Among the four extracts tested, Curcuma xanthorrhiza extract exhibited the highest cell density (1.7532) and proteolytic (12.012 U/mL), along with the lowest pH. Pineapple juice also showed a lowest pH value (5.685). The mixture extract demonstrated the highest lactic acid production (538.812% v/v), phenolics (222.494 ppm) and antioxidative capacity (IC50 2.3347). Saccharide utilization was observed for all supplemented media after an 8 h fermentation process, exceeding 85%. The findings demonstrated that all plant extracts stimulated L. paracasei growth and exhibited prebiotic properties, suggesting their potential commercialization as new functional foods and beverages.

Isolation and Identification of Chicken-Derived Lactic Acid Bacteria: In Vitro Probiotic Properties and Antagonistic Effects against Salmonella pullorum, Staphylococcus aureus, and Escherichia coli.

The development of probiotics capable of quickly colonizing the intestines of animals is important in promoting the healthy growth of livestock. The aim of this study was to screen lactic acid bacteria (LAB) from the intestinal microbiota of chickens with potential applications, and to evaluate their probiotic properties and antagonistic abilities against Salmonella pullorum, Staphylococcus aureus, and Escherichia coli. The results showed that a total of 79 strains with the characteristics of LAB were isolated from the chicken cecum microbiota, of which 7 strains exhibited strong inhibitory activity against S. pullorum, S. aureus, and E. coli. Performing 16s rDNA sequencing revealed that these seven strains were Lactiplantibacillus pentosus (n = 1), Lactiplantibacillus plantarum (n = 3), Lactiplantibacillus paraplantarum (n = 1), Lactiplantibacillus argentoratensis (n = 1), and Lactiplantibacillus fabifermentans (n = 1). Among them, L. pentosus R26 and L. plantarum R32 exhibited superior antibacterial activity. These two strains demonstrated high lactic acid production ability, with survival rates of 86.29% and 87.99% after 3 h of treatment at pH 1.5, 86.66% and 85.52% after 3 h of treatment with 0.5% bile salts, 90.03% and 88.16% after 2 h of treatment with simulated gastric fluid, and 98.92% and 98.22% after 2 h of treatment with simulated intestinal fluid, respectively. Co-cultivation with L. pentosus R26 for 24 h resulted in 50% of the pathogens being antagonized, while almost complete inhibition was observed following 72 h of co-cultivation. In conclusion, L. pentosus R26 and L. plantarum R32 exhibited high antibacterial activity and acid production capability, while also demonstrating satisfactory tolerance to low pH values and high concentrations of bile salts and digestive fluid. The probiotic characteristics and stress resistance of L. pentosus R26 were slightly superior to those of L. plantarum R32, indicating its potential for development as a probiotic.

Lactic acid production from different sources of organic solid waste: evaluation of the inoculum type and operational optimization

AbstractThis work aimed to evaluate the effect of the sources of organic solid waste on lactic acid (HLa) production. Organic fractions from cafeteria (CW), market (MW), and recycling plant (RW) wastes were used as substrates. HLa production during the self-fermentation of CW, MW, and RW, as well as during the fermentation of each substrate with anaerobic sludge (CW + sludge, MW + sludge, and RW + sludge), was evaluated in batch tests at 37 °C, initial pH of 6.5, and 150 rpm. Subsequently, the initial pH and substrate concentration of self-fermentation of CW were optimized using a central composite design. The highest HLa production was observed in the self-fermentation of CW, where several Lactobacillus species predominated in the microbial community. The higher value of HLa production using CW (20.7 g/L) was obtained at a pH of 7.29 and 115.5 gVS/L. In the optimization tests, Lactobacillus and Weissella spp. were identified as the dominant lactic acid bacteria; however, only Lactobacillus species were associated with the highest HLa production.

Metabolic Regulation-Mediated Reversion of the Tumor Immunosuppressive Microenvironment for Potentiating Cooperative Metabolic Therapy and Immunotherapy.

Tumor cells exhibit heightened glucose (Glu) consumption and increased lactic acid (LA) production, resulting in the formation of an immunosuppressive tumor microenvironment (TME) that facilitates malignant proliferation and metastasis. In this study, we meticulously engineer an antitumor nanoplatform, denoted as ZLGCR, by incorporating glucose oxidase, LA oxidase, and CpG oligodeoxynucleotide into zeolitic imidazolate framework-8 that is camouflaged with a red blood cell membrane. Significantly, ZLGCR-mediated consumption of Glu and LA not only amplifies the effectiveness of metabolic therapy but also reverses the immunosuppressive TME, thereby enhancing the therapeutic outcomes of CpG-mediated antitumor immunotherapy. It is particularly important that the synergistic effect of metabolic therapy and immunotherapy is further augmented when combined with immune checkpoint blockade therapy. Consequently, this engineered antitumor nanoplatform will achieve a cooperative tumor-suppressive outcome through the modulation of metabolism and immune responses within the TME.

NPAS2, transcriptionally activated by ARRB1, promotes the malignant behaviours of lung adenocarcinoma cells and regulates the reprogramming of glucose metabolism.

Lung adenocarcinoma (LUAD) is a serious threat to public health and is accompanied by increased morbidity and mortality worldwide. Neuronal PAS domain protein2 (NPAS2) has been confirmed as an oncogene in LUAD; however, little is known about its molecular mechanism. Here, the expression level of NPAS2 was detected in LUAD cell lines and 16HBE cells. Gain- and loss-of-function experiments were performed. Cell Counting Kit-8, colony formation, flow cytometry, wound-healing and Transwell assays were conducted to assess cell proliferation, apoptosis, migration and invasion, respectively. Reprogramming of glucose metabolism was evaluated via oxygen consumption rate (OCR), complexes activities, lactic production and glucose consumption. The expression of critical proteins was examined by western blot. We demonstrated aberrant upregulation of NPAS2 and β-arrestin-1 (ARRB1) in LUAD cell lines. ARRB1 was found to be a critical transcription factor of NPAS2 with binding sites within the promoter region of NPAS2, thereby causing its transcriptional activation. Functional experiments revealed that NPAS2 depletion significantly inhibited the malignant behaviours of A549 cells by suppressing cell proliferation, migration, invasion and epithelial-mesenchymal transition and promoting cell apoptosis. Meanwhile, NPAS2 depletion increased OCR and activities of complexes (I, II, III and V), and reduced lactic acid production and glucose uptake in A549 cells, indicating that NPAS2 depletion inhibited aerobic glycolysis, accompanied by reduced expression of glycolytic enzymes. However, the changes caused by NPAS2 knockdown were partly restored by ARRB1 overexpression. In conclusion, our study suggests that ARRB1 could transcriptionally activate NPAS2, facilitating malignant activities and glycolysis, and ultimately promoting the progression of LUAD, proving a novel therapeutic strategy for the treatment of LUAD.

Lactic Acid Production by Lactiplantibacillus plantarum AC 11S-Kinetics and Modeling.

Lactic acid is a versatile chemical with wide application in many industries. It can be produced by the fermentation of different sugars by various lactobacilli and investigations on lactic acid production from different substrates and by different strains are still in progress. The present study aimed to study lactic acid production from lactose by Lactiplantibacillus plantarum AC 11S and to choose a mathematical model describing in the best way the experimental data obtained. The influence of initial substrate concentration was investigated, and optimal pH and temperature were determined. An unstructured mathematical model was developed comprising equations for bacterial growth, substrate consumption, and product formation. The model was solved with different terms for specific growth rates considering substrate and/or product inhibition. The best bacterial growth and lactic acid production were achieved at pH = 6.5 and 30 °C. Production of lactic acid was mainly growth-associated, and at initial substrate concentration over 15 g/L, a considerable product inhibition was observed. The parameters of different models were determined and compared. The modified Gompertz equation gave the best fit when solving only the equation for biomass growth at different initial substrate concentrations. Solving the entire set of differential equations for bacterial growth, substrate consumption, and product formation, the best results were obtained when using a variant of the logistic equation for biomass growth. This variant included a term for product inhibition and described in the best way all experimental data. Solving the model for different biomass concentrations showed that an increase in biomass led to a shorter lag phase and the stationary phase was reached faster. The results obtained, optimum conditions and the kinetic model, are good bases for studying pH-controlled fermentation, as well as a continuous process.

Quality improvement of soybean meal by simultaneous microbial fermentation and enzymolysis and untargeted metabolomic analysis of its metabolites

Twelve lactobacillus strains, four yeast strains, and eight proteases were screened, and their effects on oligosaccharide degradation, lactic acid production, and proteolytic capacity were investigated in soybean meal. The simultaneous microbial fermentation and enzymolysis (SMFE) conditions were optimized as feed/water ratio of 1:0.8 (w/v), yeast inoculum of 2.1×105 CFU/g, lactobacillus inoculum of 2.1×107 CFU/g, protease addition of 200 U/g, and fermentation at 37 °C for 48 h. The results showed that SMFE soybean meal exhibited superior performance over unprocessed soybean meal, showing increases in crude protein content from 49.8% to 56.3%, acid-soluble protein content from 3.7% to 10.4%, lactic acid production from 0 to 71.5 g/kg, and a decrease in oligosaccharide content from 6.4% to 0%. The comprehensive performance of SMFE soybean meal is higher than that from enzymolysis or fermentation alone Non-targeted metabolomic analysis revealed a diversity of metabolites in fermented soybean meal produced by different processes, and 8366 detected metabolites were mainly amino acids and their metabolites. Furthermore, among the six experimental groups, the metabolites of the SMFE group, and the yeast and lactic acid bacteria fermentation groups were the most similar. This study facilitates the assessment of overall quality of soybean meal and reveals the superiority of SMFE over traditional methods in enhancing nutritional profile of soybean meal.

Microbiome-derived acidity protects against microbial invasion in Drosophila

Microbial invasions underlie host-microbe interactions resulting in pathogenesis and probiotic colonization. In this study, we explore the effects of the microbiome on microbial invasion in Drosophila melanogaster. We demonstrate that gut microbes Lactiplantibacillus plantarum and Acetobacter tropicalis improve survival and lead to a reduction in microbial burden during infection. Using a microbial interaction assay, we report that L.plantarum inhibits the growth of invasive bacteria, while A.tropicalis reduces this inhibition. We further show that inhibition by L.plantarum is linked to its ability to acidify its environment via lactic acid production by lactate dehydrogenase, while A.tropicalis diminishes the inhibition by quenching acids. We propose that acid from the microbiome is a gatekeeper to microbial invasions, as only microbes capable of tolerating acidic environments can colonize the host. The methods and findings described herein will add to the growing breadth of tools to study microbe-microbe interactions in broad contexts.

Herbal Composite Preparation and Investigating its Efficiency to Inhibit Biofilm Formation and Virulence Factors of Prevotella Intermedia and Porphyromonas Gingivalis – Formulation of Mouthwash Using a Herbal Composite and Evaluating its Anti-microbial Activity

ABSTRACT Herbal composite preparation was studied with the aim of inhibiting the virulence factors of two dental pathogens: Prevotella intermedia and Porphyromonas gingivalis. A novel herbal composite was developed using the herbal extracts of Wrightia tinctoria and Bauhinia variegata. During the study, the following observations were noted. The minimal inhibitory concentration of Wrightia tinctoria and Bauhinia variegata composites (WBc) was obtained for the test concentration of 20 μg/ml (16 ± 0.57 mm and 15 ± 0.75 mm of inhibitory zones against Prevotella intermedia and Porphyromonas gingivalis, respectively). Biofilm inhibition assay results revealed about 0.51 ± 1.25 mg/ml and 0.53 ± 0.57 mg/ml of minimal biofilm eradication concentration (MBEC) against Prevotella intermedia and Porphyromonas gingivalis, respectively. The effect of WBc on lactic acid production showed that 200 μg/ml and 400 μg/ml concentrates reduced up to 80% and 70% in Prevotella intermedia and Porphyromonas gingivalis, respectively. Formulated herbal mouthwash showed good stability under all three different test conditions (5°C, 25°C, and 40°C) as the color, odor, phase separation, and homogeneity were not changed for the period of 3 months. The anti-bacterial activity of formulated mouthwash (30 μg/ml) exhibited maximum inhibitory zones of about 18 ± 0.75 mm and 19 ± 1.05 mm against the respective test bacteria – Prevotella intermedia and Porphyromonas gingivalis. Amplification of mfa1 and clpB genes showed 246 bp and 294 bp fragments of P. gingivalis and 238 bp and 280 bp fragments of P. intermedia during agarose electrophoretic analysis. The docking report revealed -5.84 Kcal/Mol binding energy and found three hydrogen bonding between the quercetin and target protein, mfa1 of Porphyromonas gingivalis. The target protein, clpB of Prevotella intermedia, and quercetin had -6.72 Kcal/Mol binding energy and found four hydrogen bonds between them. The developed composite could be optimized in future to develop a novel and biocompatible herbal mouthwash for the prevention of different dental caries and gingival inflammation associated with dental biofilm formation.