Optimization Of Fermentation Process Research Articles

Exploring Calcium Alginate-Based Gels for Encapsulation of Lacticaseibacillus paracasei to Enhance Stability in Functional Breadmaking

This study focuses on evaluating the efficiency of acid-tolerant Lacticaseibacillus paracasei bacteria encapsulated in an alginate-based gel matrix during repeated sourdough fermentation cycles, as well as their preservation during storage and throughout baking at high temperature. A double-coating procedure was applied, involving the encapsulation of bacterial cells in calcium alginate, which was further coated with chitosan. The encapsulation efficiency (EE) did not show significant difference between alginate and alginate–chitosan (97.97 and 96.71%, respectively). The higher number of L. paracasei bacteria was preserved in double-coated microbeads, with survivability rates of 89.51% and 96.90% in wet and dried microbeads, respectively. Encapsulated bacteria demonstrated effective fermentation ability, while double gel-coated cells exhibited slower acidification during sourdough fermentation, maintaining higher efficiency in the second fermentation cycle. The addition of freeze-dried, alginate-based gel-encapsulated bacteria (2–4%, w/w flour) significantly (p < 0.05) improved bread quality and extended its shelf life. A double-layer coating (alginate–chitosan) can be introduced as an innovative strategy for regulating the release of lactic acid bacteria and optimizing fermentation processes. Powdered alginate or alginate–chitosan gel-based L. paracasei microcapsules, at appropriate concentrations, can be used in the production of baked goods with acceptable quality and sensory properties, achieving a lactic acid bacteria count of approximately 106 CFU/g in the crumb, thereby meeting the standard criteria for probiotic bakery products.

Waste to wealth: Polyhydroxyalkanoates (PHA) production from food waste for a sustainable packaging paradigm

The growing demand for sustainable food packaging and the increasing concerns regarding environmental pollution have driven interest in biodegradable materials. This paper presents an in-depth review of the production of Polyhydroxyalkanoates (PHA), a biodegradable polymer, from food waste. PHA-based bioplastics, particularly when derived from low-cost carbon sources such as volatile fatty acids (VFAs) and waste oils, offer a promising solution for reducing plastic waste and enhancing food packaging sustainability. Through optimization of microbial fermentation processes, PHA production can achieve significant efficiency improvements, with yields reaching up to 87 % PHA content under ideal conditions. This review highlights the technical advancements in using PHA for food packaging, emphasizing its biodegradability, biocompatibility, and potential to serve as a biodegradable alternative to petroleum-based plastics. However, challenges such as high production costs, mechanical limitations, and the need for scalability remain barriers to industrial adoption. The future of PHA in food packaging hinges on overcoming these challenges through further research and innovation in production techniques, material properties, and cost reduction strategies, along with necessary legislative support to promote widespread use.

Xylitol production from passion fruit peel hydrolysate: Optimization of hydrolysis and fermentation processes

The passion fruit peel (PFP) has a high cellulose and hemicellulose content, which can be used to produce fermentable sugars. In this context, this study aims to optimize the release of xylose and the production of xylitol from PFP. The optimized conditions were 0.71 M dilute sulfuric acid and a 21.84-minute treatment, yielding 19.03 g/L of xylose (PFP-1). Different PFP hydrolysates were evaluated to improve xylitol production by the yeast Kluyveromyces marxianus ATCC 36907: PFP-2 (PFP1 treated with Ca(OH)2), PFP-3 (PFP-1 treated with Ca(OH)2 and activated carbon), PFP-4 (PFP-3 with biological elimination of glucose with S. cerevisiae, and concentrated at different xylose concentrations). The applied methods resulted in higher xylitol production (14.97 g/L), when PFP hydrolysate was detoxified with Ca(OH)2, treated with activated charcoal for 1 h, biotreated for glucose removal, and concentrated to 40 g/L of xylose.

Production of Amylase by Solid State Fermentation Using Agricultural Waste

This study presents a comprehensive investigation into the production of amylase, a crucial enzyme with wide-ranging industrial applications, using locally sourced substrates from Kachchh, Gujarat. The research employed the Bacillus licheniformis strain and substrates such as coconut, rice husk, wheat bran, paddy straw, and maize straw. The study found paddy straw to be the most promising substrate for amylase production. The research also systematically optimized various process parameters for amylase production in Solid-State Fermentation (SSF) using the One Variable at a Time (OVAT) method. These parameters included incubation period, temperature, inoculum level, additional carbon sources, starch concentrations, additional nitrogen sources, initial pH, different mineral salt ions, initial moisture level, and surfactants. The results showed that the optimal conditions for maximum amylase yield were an incubation period of 48 hours, an incubation temperature of 35°C, an inoculum level of 10%, starch as the additional carbon source, a starch concentration of 2.5%, yeast extract as the additional nitrogen source, an initial pH of 7, NaCl as the mineral salt, an initial moisture level of 75%, and Tween 80 as the surfactant. This research provides a reliable and sustainable approach to enzyme production, offering valuable insights for the optimization of the solid-state fermentation process for maximum amylase production.

Effects of tomato straw fermentation on nutrients and bacterial community structure

Unsustainable straw treatment methods detrimentally affect the environment and ecology. Aerobic fermentation (AE) and anaerobic fermentation (AN) are environmentally friendly treatments that better utilise straw resources. In this study, high-throughput sequencing was used to investigate the effects of AE and AN on nutrient content and microbial community structure during tomato straw fermentation. Nitrate nitrogen, available phosphorus, available potassium, and fulvic acid contents following AE were 1250.04 mg/kg, 80.34 %, 161.39 %, and 49.31 %, respectively, which were higher than those following AN. Ammonium nitrogen, humic acid, and humic substance levels following AN were 309.07 %, 31.18 %, and 17.38 %, respectively, which were higher than those following AE. Firmicutes (24.76 %) and Actinobacteria (12.93 %) were more abundant following AE, whereas Proteobacteria (33.82 %) and Bacteroidetes (33.82 %) exhibited higher abundance following AN. AE more effectively eliminated pathogenic bacteria (22.01%–0.26 %) and encouraged stronger interactions between dominant bacterial genera. Redundancy and Mantel test analyses revealed that electrical conductivity and temperature were the most important environmental factors affecting bacterial communities in AE and AN, respectively. AE had a stronger effect on effective nutrient release from tomato straw, implying its greater application potential as a fertiliser. Overall, our study provides a theoretical basis for the optimisation of fermentation methods and processes.

Utilization of citrus, date, and jujube substrates for anaerobic digestion processes

AbstractThis research explores the potential for generating biogas and clean energy by processing organic waste, a process that can become a sustainable solution to Syria's energy needs. Focusing on agricultural residues generated from citrus fruit orange, date, and jujube cultivation in Syria, this study evaluates the potential for anaerobic digestion of these residues for biogas production. It highlights the influence of substrate composition and the optimization of fermentation processes on biogas and methane production. The study focuses on evaluating the anaerobic digestion process by examining various dosages ranging from 20% to 50% dry matter for citrus orange waste, and different types of substrate with a fixed ratio of 20% substrate dry matter. It specifically discusses the factors influencing the inhibitory effect of anaerobic digestion, giving particular consideration to orange waste, a significant byproduct of the citrus industry. The biogas produced maintained a stable methane content when a citrus‐to‐inoculum ratio of 30:70 was used. Jujube waste, characterized by a composition rich in cellulose and hemicellulose, exhibited a higher potential for biogas and methane generation among the fruit waste investigated, particularly when combined with the inoculum in a 20:80 ratio. The research findings underscore the potential of using Syrian agricultural residues, including orange citrus peel, date, and jujube fruit, for the production of biogas through anaerobic digestion.

Impact of Bacillus subtilis on Chinese yellow rice wine (Huangjiu) fermentation: Method variations and flavor analysis

This investigation explores the impact of various fermentation techniques and the inoculation of Bacillus subtilis spores on the physicochemical properties and principal flavor profiles of Huangjiu. Employing sensory analysis, headspace solid-phase microextraction, gas chromatography-tandem mass spectrometry (HS-SPME-GC–MS), and orthogonal partial least squares discriminant analysis (OPLS-DA), we observed that these variables significantly alter the physicochemical attributes of Huangjiu. Our analysis, integrating volatile organic compounds (VOCs) with odor activity values (OAV), revealed that while B. subtilis inoculation modifies the concentrations of key flavor compounds, it does not affect their types. Notably, the inoculation enhances the concentrations of 13 primary flavor compounds, thereby enriching floral and fruity notes while reducing higher alcohol levels. These findings contribute valuable insights into the flavor formation mechanisms of Huangjiu and guide the optimization of fermentation processes.

A Preliminary Study on the Effect of Adding Sugarcane Syrup on the Flavor of Barley Lager Fermentation.

This study focuses on the diversified utilization of the sugarcane industry, and sugarcane syrup, as a by-product of the sugarcane industry, is a good raw material for fermentation. Bringing sugarcane syrup into beer is conducive to the enrichment of the sugar industry, and it can improve the flavor of beer and make it more aromatic. This study determined the optimal fermentation process for beer. By analyzing the consumption rate of the carbon and nitrogen sources of raw materials, the nutrient utilization of yeast, and the causes of differences in flavor substances, the flavor composition and flavor stability of beer were determined by SPME-HS-GC-MS technology. The results showed that beer brewed with sugarcane syrup as an auxiliary raw material met the basic specifications of beer. The addition of sugarcane syrup to the wort base increased the utilization of amino acids by the yeast, and LS (lager with added cane syrup) increased the nine flavor compounds of the beer, which constituted the basic flavor of the beer, bringing new flavor compounds compared with the normal all-barley beer. Forced aging experiments showed that LS produced fewer aging compounds than OWBL. Various experiments have shown that it is feasible to ferment beer with sugarcane syrup instead of partial wort.

Online Monitoring of the Temperature and Relative Humidity of Recycled Bedding for Dairy Cows on Dairy Farms

In large-scale dairy farming, the use of high-temperature-fermented dairy manure bedding instead of rice husk-based bedding and other commercial types of bedding is widely favored. Strip-stacking aerobic fermentation is the main production method of dairy manure bedding, but it has problems including unstable fermentation and the secondary breeding of pathogens. In this work, a multi-probe, integrated, online monitoring system for temperature and relative humidity was used for fermentation process optimization. The effects of the temporal and spatial distribution of fermentation temperature and relative humidity on the nutrient content curve and the moisture and ash content of manure bedding materials were systematically studied. The effect of the fermentation process on the retention rate of effective bedding materials (cellulose, hemicellulose, and lignin) was analyzed. The experiments proved that high-quality bedding material can be obtained through reasonable stacking fermentation. The fabricated bedding material has a total dry base content consisting of cellulose, hemicellulose, and lignin of 78%, an ash content of 6%, and a nutrient content of 17%. The obtained bedding material was produced to increase the bed rest rate and continuously inhibit the bedding bacteria content, keeping it at a low level for 5 days. This study proves that temperature and humidity monitoring can guide the optimization of the strip-stacking fermentation process of dairy manure and that it can be applied to large-scale farms to improve fermentation parameters.

Enhancing the yield of Xenocoumacin 1 in Xenorhabdus nematophila YL001 by optimizing the fermentation process

Xenocoumacin 1 (Xcn 1), antibiotic discovered from secondary metabolites of Xenorhabdus nematophila, had the potential to develop into a new pesticide due to its excellent activity against bacteria, oomycetes and fungi. However, the current low yield of Xcn1 limits its development and utilization. To improve the yield of Xcn1, response surface methodology was used to determine the optimal composition of fermentation medium and one factor at a time approach was utilized to optimize the fermentation process. The optimal medium composed of in g/L: proteose peptone 20.8; maltose 12.74; K2HPO4 3.77. The optimal fermentation conditions were that 25 °C, initial pH 7.0, inoculum size 10%, culture medium 75 mL in a 250 mL shake flask with an agitation rate of 150 rpm for 48 h. Xenorhabdus nematophila YL001 was produced the highest Xcn1 yield (173.99 mg/L) when arginine was added to the broth with 3 mmol/L at the 12th h. Compared with Tryptic Soy Broth medium, the optimized fermentation process resulted in a 243.38% increase in Xcn1 production. The obtained results confirmed that optimizing fermentation technology led to an increase in Xcn1 yield. This work would be helpful for efficient Xcn1 production and lay a foundation for its industrial production.

Isolation, purification, characterization, and analysis of the antioxidant activity of antioxidant peptides from walnut milk fermented with Lactobacillus paracasei SMN-LBK

Microbial fermentation is an important method to obtain antioxidant peptides. In this study, Lactobacillus paracasei SMN-LBK was used to ferment biprotein (walnut milk) to enhance the antioxidant capacity of the active peptide. The optimal fermentation process of 5% inoculum, 1:7 plant protein ratio, 37 °C and 12 h resulted in 77.63% ± 0.36% DPPH scavenging, 93.31% ± 0.25% ABTS scavenging and 79.37% ± 0.02% metal ion chelating capacity. The hydrolysis products were classified into three molecular weight classes using an ultrafiltration membrane system. DPAP-3 with a molecular weight of less than 3 kDa had the highest antioxidant activity. DPAP-3 was further separated into three subclasses using gel filtration chromatography. These subclasses were characterized using nano-liquid chromatography-tandem mass spectrometry (NanoLC-MS/MS) and screened by searching https://biochemia.uwm.edu. Four antioxidant peptides were identified using a combination of hydrophobic amino acids, positive ion-charge interactions, and numerical values. Two of these peptides were from animals and one from plants. Their amino acid sequences are as follows PCRGVLLR, IHIPLPRWV, QSKVLPVPQK, and KVLPVPQKA, with molecular docking binding energies of −7.8, −8.3, −7.4, and −7.7 kcal/mol, respectively. The results suggest that fermentation of plant-animal proteins with Lactobacillus paracasei can produce antioxidant peptides that can be used as functional ingredients in food products. This has the potential to conserve animal protein resources.

A machine learning-based approach for improving plasmid DNA production in Escherichia coli fed-batch fermentations.

Artificial Intelligence (AI) technology is spearheading a new industrial revolution, which provides ample opportunities for the transformational development of traditional fermentation processes. During plasmid fermentation, traditional subjective process control leads to highly unstable plasmid yields. In this study, a multi-parameter correlation analysis was first performed to discover a dynamic metabolic balance among the oxygen uptake rate, temperature, and plasmid yield, whilst revealing the heating rate and timing as the most important optimization factor for balanced cell growth and plasmid production. Then, based on the acquired on-line parameters as well as outputs of kinetic models constructed for describing process dynamics of biomass concentration, plasmid yield, and substrate concentration, a machine learning (ML) model with Random Forest (RF) as the best machine learning algorithm was established to predict the optimal heating strategy. Finally, the highest plasmid yield and specific productivity of 1167.74mg L-1 and 8.87mg L-1/OD600 were achieved with the optimal heating strategy predicted by the RF model in the 50 L bioreactor, respectively, which was 71% and 21% higher than those obtained in the control cultures where a traditional one-step temperature upshift strategy was applied. In addition, this study transformed empirical fermentation process optimization into a more efficient and rational self-optimization method. The methodology employed in this study is equally applicable to predict the regulation of process dynamics for other products, thereby facilitating the potential for furthering the intelligent automation of fermentation processes.

Improvement of nucleotide content of Cordyceps tenuipes by Schisandra chinensis: fermentation process optimization and application prospects.

Nucleotides are important components and the main indicators for judging Cordyceps quality. In this paper, the mixed fermentation process of Schisandra chinensis and Cordyceps tenuipes was systematically studied, and it was proposed that the fermentation products aqueous extract (S-ZAE) had antioxidant activity and anti-AChE ability. Herein, the results of a single factor showed that S. chinensis, yeast extract, inoculum amount, and pH had significant effects on nucleotide synthesis. The fermentation process optimization results were 3% glucose, 0.25% KH2PO4, 2.1% yeast extract, and S. chinensis 0.49% (m/v), the optimal fermentation conditions were 25℃, inoculum 5.8% (v/v), pH 3.8, 6 d. The yield of total nucleotides in the scale-up culture was 0.64 ± 0.027mg/mL, which was 10.6 times higher than before optimization. S-ZAE has good antioxidant and anti-AChE activities (IC50 0.50 ± 0.050mg/mL). This fermentation method has the advantage of industrialization, and its fermentation products have the potential to become good functional foods or natural therapeutic agents.

Optimization of fermentation process and characterization of non-alcoholic functional beverage from pigmented rice varieties

Traditional foods have recently gained importance because of their role in the United Nations’ sustainable development goals (SDG), ensuring health benefits and economic development. Fermented rice water (FRW) is a popular traditional drink among low socio-economic groups, especially in Southeast Asia. This study focused on optimizing fermentation conditions for sustainable food production and consumption of FRW using pigmented rice, followed by analyzing the nutritional, physicochemical and phytochemical properties. Fermenting cooked rice in water for 16 h at 37 °C produced bubbles, mild sourness and zero alcohol content. Flavors and sugar were added to optimized FRW as food additives to enhance palatability. Nutritional profiling showed that black flavored fermented rice water (FFRW) had the highest protein (0.06%), carbohydrate (13.25 g/100 mL) and calorific value (78.09 kcal/100 mL). However, brown FFRW had the highest vitamin B3 (3.12 mg/L) and mineral content. Phenolic and antioxidant activity was highest in black FFRW, followed by red and brown, with maximum bio-accessibility of polyphenols found in black FFRW (75.25%). All FFRW were microbiologically safe and stable during a storage period of 30 days at 4 °C. All FRW had >8 log CFU count, with black FFRW having the highest LAB count, indicating their possible probiotic potential in addressing malnutrition and undernourishment. Thus, FFRW holds the potential to be commercialized in the food and beverage sector due to its rich nutritional and functional properties. Furthermore, the consumption of FFRW will preserve traditional knowledge, ensure sustainable livelihoods and contribute to achieving development goals.

Characterization and optimization of gellan gum production by natural Sphingomonas sp. SM2

Gellan is one of the most important industrial microbial exopolysaccharides used for a wide range of applications in food, pharmaceutical, and oil. In this study, a native strain of Sphingomonas producing gellan gum was identified from rice roots. Various factors, including carbon and nitrogen sources, pH levels, fermentation duration, and aeration, were investigated to enhance the production of gellan by Sphingomonas sp. SM2. The highest efficiency in gum production was achieved by a medium composed of 30 g/L glucose. When the carbon-to-nitrogen ratio was 30, organic nitrogen caused this increase. By incorporating all relevant factors, the optimized fermentation process can yield a gum concentration of 8.64 ± 0.34 g/L. The total carbohydrate content of the native gum was found to be 52.62 ± 0.52%. The existence of mucilage surrounding the bacterial cells was verified by scanning electron microscopy images. The extracted gellan gum was characterized using FTIR, 1H NMR, and HPLC analyses. The structure of the gum was confirmed by comparing it with the standard gellan gum. The production of gellan gum by Sphingomonas sp. SM2 using the utilization of diverse carbon sources suggests its immense potential as a gellan producer.

Microbial chassis design and engineering for production of gamma-aminobutyric acid.

Gamma-aminobutyric acid (GABA) is a non-protein amino acid which is widely applied in agriculture and pharmaceutical additive industries. GABA is synthesized from glutamate through irreversible α-decarboxylation by glutamate decarboxylase. Recently, microbial synthesis has become an inevitable trend to produce GABA due to its sustainable characteristics. Therefore, reasonable microbial platform design and metabolic engineering strategies for improving production of GABA are arousing a considerable attraction. The strategies concentrate on microbial platform optimization, fermentation process optimization, rational metabolic engineering as key metabolic pathway modification, promoter optimization, site-directed mutagenesis, modular transporter engineering, and dynamic switch systems application. In this review, the microbial producers for GABA were summarized, including lactic acid bacteria, Corynebacterium glutamicum, and Escherichia coli, as well as the efficient strategies for optimizing them to improve the production of GABA.

Optimization of watermelon peel yogurt fermentation process and study on antioxidant capacity

This study aims to explore the variation patterns of total flavonoid and other antioxidant activities in watermelon peel yogurt, providing a basis for the production of a functional yogurt. Using watermelon peel as the raw material and total flavonoid content as the indicator, the optimal process of watermelon peel yogurt was optimized through single-factor and orthogonal experiments. The results indicate that yogurt fermented with a mass ratio of watermelon juice to milk at 1:3, fermentation time of 10 hours, and fermentation agent dosage of 0.10% exhibits a refreshing watermelon aroma, excellent texture, and almost no whey separation. After process optimization, the polysaccharide content of watermelon peel yogurt increased by 19%, and water-holding capacity rose to 63.3%. Texture analysis revealed a hardness of 81.50 g, viscosity of 20.00 g, adhesiveness of 1.20 mJ, elasticity of 8.83 mm, and a draw length of 6.76 mm. The physicochemical and microbiological properties of watermelon peel yogurt meet national standards, and it possesses a good free radical scavenging ability.

Evaluation of stress tolerance and design of alternative culture media for the production of fermentation starter cultures in cacao

Ecuador is one of the world's leading producers of cacao beans, and Nacional x Trinitario cacao represents one of the most distinctive varieties due to its flavor and aroma characteristics. This study aimed to evaluate the effect of the starter culture isolated from microbial diversity during the spontaneous fermentation of Nacional x Trinitario cacao. A total of 249 microbial isolates were obtained from spontaneous culture, with Lactiplantibacillus (45 %), Saccharomyces (17 %), and Acetobacter (2 %) being the most relevant genera for fermentation. Tolerance tests were conducted to select microorganisms for the starter culture. Lactiplantibacillus plantarum exhibited the highest tolerance at pH 5 and 6 % ethanol and tolerated concentrations up to 15 % for glucose and fructose. Acetobacter pasteurianus grew at pH 2 and 6 % ethanol, tolerating high sugar concentrations of up to 15 % for glucose and 30 % for fructose, with growth observed in concentrations up to 5 % for lactic and acetic acid. Subsequently, a laboratory-scale fermentation was conducted with the formulated starter culture (SC) comprising S. cerevisiae, L. plantarum, and A. pasteurianus, which exhibited high tolerance to various stress conditions. The fermentation increased alcoholic compounds, including citrusy, fruity aromas, and floral notes such as 2-heptanol and phenylethyl alcohol, respectively 1.6-fold and 5.6-fold compared to the control. Moreover, the abundance of ketones 2-heptanone and 2-nonanone increased significantly, providing sweet green herbs and fruity woody aromas. Cacao fermented with this SC significantly enhanced the favorable aroma-producing metabolites characteristic of Fine-aroma cacao. These findings underscore the potential of tailored fermentation strategies to improve cacao product quality and sensory attributes, emphasizing the importance of ongoing research in optimizing fermentation processes for the cacao industry.

Optimization of the fermentative production of L-methionine by engineered Escherichia coli

As the only essential amino acid containing elemental sulphur, L-methionine has important physiological and biochemical functions in living organisms. However, the fermentative production of L-methionine has not met the requirements of industrial production because of its low production level. In this paper, the fermentation process of an efficient L-methionine producing strain E. coli W3110ΔIJAHFEBC trc-fliY trc-malY/PAM glyA-22 metF constructed previously was systematically optimized. Based on the optimal initial glucose concentration, the effects of different fed-batch fermentation processes, including DO-Stat, pH-Stat, controlling residual sugar control at different level and feeding glucose with constant rate, on L-methionine fermentation were studied. It was found that the control of glucose concentration greatly affected the fermentation process. Subsequently, an optimal fed-batch fermentation process was developed, where the L-methionine titer was increased to 31.71 g/L, the highest yield reported to date, while the fermentation time was shortened to 68 h. Meanwhile, a fermentation kinetics model under the optimal fed-batch fermentation conditions was established, which fitted well with the biosynthesis process of L-methionine. This study may facilitate further development of the fermentative production of L-methionine.

Screening and Optimization of α-Glucosidase Inhibitor Production by Potent Strain of Bacillus subtilis Isolated from Peruyaan, Fermented Soy-Food of Northeast India

Background. Fermented soy foods exhibit the capability to inhibit the α-glucosidase enzyme. The bacteria isolated from these fermented soy foods may contribute to the production of a higher quantity of α-glucosidase inhibitor (α-GI) in optimized condition. Aim. The present study aims to isolate α-glucosidase inhibitor producing bacterial strain from peruyaan, a traditional fermented soy food of Arunachal Pradesh, optimize the production of α-GI for maximum yield, and assess the compounds. Result. Bacillus subtilis PM NEIST_4 (identified by 16S rRNA gene sequencing) was isolated as a potential strain that exhibit maximum α-glucosidase inhibition. This strain showed maximum α-GI productivity in Mueller Hinton Broth (MHB). The cultivation parameters for fermentation of MHB were optimized in order to maximize the yield of α-GIs. The α-GI productivity of 1616.613 ± 84.54 U/mL was observed before optimization, which increased by almost three times after optimization (4808.324 ± 13 U/mL). In vitro assays against α-glucosidase enzyme revealed a significant IC50 value of 596.532 ± 44.80 μg/mL after the optimized study compared to that before optimization (IC50 1705.617 ± 43.95 ug/mL). Besides, α-glucosidase enzyme inhibitory property of fermented MHB (FMHB) was found to be unaffected at varied range of pH and temperature. Oral gavaging of FMHB in sucrose-loaded Wistar rat exhibited maximum reduction in postprandial blood glucose at 400 mg/kg body weight. Furthermore, FMHB was fractionated using solvents of increasing polarity including n-hexane, ethyl acetate, and n-butanol. Ethyl acetate fraction that exhibited the strongest inhibitory action against the α-glucosidase enzyme (IC50 value of 251.55 ± 19.65 μg/mL), showed the presence of pyrrolopyrazine derivatives through GC-MS and LC-MS analyses. Conclusion. The present study demonstrated the isolation of a potent α-GI producing bacterial strain from traditional fermented soy food, and the optimization of the bacterial fermentation process was achieved to maximize the yield of α-GI. These findings underscore promising industrial potential of B. subtilis PM NEIST_4, advocating the use of indigenous microbial resources for the production of α-GI.