Amylase Production Research Articles

Cost-Effective Strategy and Feasibility for Amylase Production from Okara by Bacillus subtilis J12

Low-cost enzyme production is considered a feasibility factor in enzyme commercialization. Okara, a high-nutritional agro-industrial residue from soybean processing, was performed as a medium for bacterial amylase production to save costs and increase productivity. This study aimed to produce, characterize, activate amylase, and evaluate the material cost for media from okara. Under solid-state fermentation (SSF) of okara without pretreatment, Bacillus subtilis J12 could produce 983 U/g of amylase within 24 h. Bacillus subtilis J12 amylase had optimal activity at pH 6.0 and 50 °C and was stable at a moderate temperature for up to 120 min. Identified as a metalloenzyme, the activity was improved by ferric ions. The purification of amylase resulted in two fractions which contained at least two types of amylases. Compared with other producers, the production was evaluated using low-cost media without additional supplementations. Based on the productivity, characteristics, and evaluation, Bacillus subtilis J12 amylase was potentially commercialized, had economic value, possessed energy-saving features, and could be applied for industrial use.

TEMPORARY REMOVAL: Bioprospecting amylase from Samiti Lake, situated in the eastern Himalayas

Enzymes, especially amylases, have been an economic boon to the industrial sector, their bioprospective and biotechnological use is an added advantage. Our primary focus of the study was to isolate the most potent amylase producer and to optimize its production parameters through One Factor At A Time (OFAT), Central Composite Rotatable Design Response Surface Methodology (CCRD RSM) and Artificial Neural Network (ANN). Based on the qualitative and quantitative analysis, SLAB1 was selected as the most potent amylase producer out of the potential isolates. Further SLAB1 was identified as Priestia flexa via 16SrRNA identification protocol. Optimization of the production parameters showed the best carbon, nitrogen sources, temperature and pH to be fructose, peptone, 20 °C and pH 8.0 respectively. Further, the enzyme was purified using ammonium sulphate precipitation followed by dialysis. Later, DEAE Sepharose (Sigma) resin was used for ion exchange chromatography and the protein was eluted using NaCl gradients from 0.1 M – 0.6 M. Enzyme kinetics assessment of the purified amylase with the Lineweaver Burk plot showed values of maximum rate; Vmax (10.869 μmoL/min), and Michaelis-Menten constant Km to be around (14.91 mg/mL). To determine its potential application, analysis of this purified amylase in cleaning the tomato and chocolate stained cotton fabrics after comparing its compatibility with different detergents were executed. Further analysis of the washed stained fabrics via Scanning Electron Microscopy was carried out.

Isolation, Characterization and Molecular Identification of Bacteria Associated with Green and Brown Seaweeds

Aims: The co-existence of bacterial communities along with macroalgae in marine environment develops a mutualistic relationship resulting in functional advantages for both the groups. The present research studied the composition of epiphytic bacteria associated with green (Ulva spp.) and brown seaweeds (Sargassum spp. and Padina spp.) sampled off Maharashtra coast, India, and their physiological and bioactive properties. Study Design: Green and brown seaweeds were collected from three locations along Maharashtra coast and analyzed for the associated bacteria. Place and Duration of Study: Seaweed samples analyzed in this study were from Kelwa, Manori and Ratnagiri along the Maharashtra coast. The analysis was performed during June to September 2021. Methodology: Bacteria from freshly collected seaweed samples were homogenized in saline, 10-fold serially diluted and plated on Zobell marine agar. Distinct colonies were selected and identified by a series of biochemical tests, followed by partial 16SrRNA gene sequencing. The physiological characteristics of the isolated bacteria were studied by screening for temperature and salinity tolerance, production of protease, lipase, agarose, amylase and biosurfactants. Results: The total seaweed bacterial count ranged from 4.5 ×103 to 2.9 × 104 CFU/g. Seventy-seven bacteria were isolated, 44 (57.14%) and 33 (42.85%) isolates from green and brown seaweeds respectively. The 16SrRNA sequencing of 20 representative isolates revealed the dominance of Bacillus spp., followed by Vibrio spp. Growth at 0°C was exhibited by all bacteria except Bacillus tequilensis, Bacillus altitudinis, Oceanobacillus iheyensis and one isolate of Vibrio spp. A majority of the isolates grew at 45°C. Vibrio spp. exhibited protease, amylase, gelatinase, and agarase activities, whereas the biosurfactant activity was commonly associated with Bacillus spp. Conclusion: The results of this study illustrate the occurrence of seaweed-associated beneficial bacteria exhibiting bioactive properties with potential biomedical applications.

Chemical Compound Identification of Lactic Acid Bacteria (LAB) From Porang Tubers Waste (A. muelleri blume) as Probiotic

The production of amylase produced by lactic acid bacteria (LAB) has better stability. The purpose of this study was to utilize and determine the characteristics of LAB isolates from porang skin waste which have the potential as probiotics and the activity of amylase enzymes. Isolation was done by fermentation method for 48 h with the addition of starter Pediococcus acidilactici that had been cultured. The results showed that the number of colonies of N1 isolates was 49 ×108 CFU/mL with regular small round colonies, having a convex elevation, the edges of the colonies were round, yellowish white, and Gram positive bacteria because they produced a purple color on Gram staining. Acid resistance test at pH 2.0; 3.0; 4.0; 5.0 and 6.0, the isolates N1 was able to survive at each pH because it showed turbidity in the growth medium. The antimicrobial activity test showed a strong zone of inhibition on S. aureus and S. pyogenes of 10,33 and 11,17 mm and a moderate zone of inhibition on K. pneumoniae, S. dysentriae, and P. aeruginosa, respectively, which was 6,33; 7,7 and 7,5 mm. The positive N1 isolate contained alkaloids as bioactive compounds from the results of phytochemical screening and the results of the FTIR analysis showed that specific groups of alkaloids were N-H, C-N, and C=O at wave numbers 3423.30 cm-1; 1235.45 cm-1; and 1634.81 cm-1.

Harnessing Bacillus subtilis QY5 PP784163 for Bioethanol Production from Potato Peel Waste and Nutrient Recovery for Animal Feed: Maximizing Resource Efficiency

The present work focuses on the utilization of potato peel waste for the production of bioethanol. In the present study, extensive screening was undertaken to isolate amylolytic and cellulolytic microbes using starchy biomass. After confirming the chemical composition of potato peel waste (PPW), several trials were performed to enhance the amylase and cellulase production from Bacillus subtilis to hydrolyze the PPW in submerged fermentation. Optimization of physical parameters was performed using both commercial and indigenous media from enzymatically hydrolyzed PPW. Different routes of various combinations were designed to enhance bioethanol production. The maximum ethanol titer of 0.50% and 0.41% was recorded in Route B and A, i.e., separate saccharification and ethanol fermentation and consolidated fermentation. Simultaneous saccharification and fermentation (SSF) also measured a good ethanol yield of 0.46%. The fermented residual cake was checked for nutritional components and showed a high content of protein and amino acids because of the addition of unicellular yeasts. This cake can be utilized as an animal feed supplement.

Genetic Diversity and Functional Potential of Streptomyces spp. Isolated from Pachmarhi Biosphere Reserve, India.

Streptomyces is a diverse genus, well known for producing a wide array of metabolites that have significant industrial utilization. The present study investigates the genetic and functional diversity of Streptomyces spp. isolated from the Pachmarhi Biosphere Reserve (PBR), India, an unexplored site. The 16S rRNA gene sequencing and analysis revealed 96 isolates belonging to 40 different species indicating a substantial phylogenetic diversity. The strains were clustered into two groups: a major cluster with 94 strains and a small cluster with two strains. BOX- PCR analyses revealed an incredible genetic diversity existing among the strains of Streptomyces spp. in PBR. The analyses revealed the intra-species diversity and inter-species closeness within the genus Streptomyces in the study area. Qualitative screening for enzyme production has shown that 53, 42, 41, 11, and 54 strains tested positive for CMCase, xylanase, amylase, pectinase, and β-glucosidase, respectively. Additionally, 54 strains tested positive for PHB production. The strains were assayed quantitatively for the production of CMCase, xylanase, amylase, and pectinase. Streptomyces sp. MP9-2, Streptomyces sp. MP10-11, Streptomyces sp. MP10-18, and Streptomyces sp. MP10-6 recorded maximum CMCase (0.604 U/mL), xylanase (0.553 U/mL), amylase (1.714 U/mL), and pectinase (13.15 U/mL) activities, respectively. Furthermore, several strains demonstrated plant growth-promoting traits, viz. zinc and phosphate solubilization and production of ammonia, HCN (hydrogen cyanide), and IAA (Indole acetic acid), and nitrogen fixation. Fifty strains showed antifungal activity against Fusarium oxysporum f. sp. lycopersici with inhibitions ranging from 7.5 to 47.5%. Current findings underscore the ecological and biotechnological significance of Streptomyces spp. in the unexplored habitat of PBR.

Improvement of thermostable productivity ?-amylase from local isolate Bacillus licheniformis H14.

(28)Bacterial local isolates of Bacillus sp. were obtained from soil samples. Isolates were tested for thermostable alpha- amylase production on solid media; fifteen isolates were able to develop clear zone around the bacterial growth after floating the plates with iodine reagent (Lugol's solution). There were further tested in submerged culture which led to selection of Bacillus sp. H14since it was the most efficient .Microbial and biochemical tests showed that the local isolate Bacillus sp.H14was refered to the species B.licheniformis that signed as H14 was refered to the species B.licheniformis H14 .,To get ahigher yield of alpha – amylase(48.70unit/mg protein) production from the local isolate B.licheniformis H14 . This study used different mutation ways such as physical way by using the physical mutagen (ultraviolet light) and chemical way by using the chemical mutagen (nitrosoguanidine). Physical mutation results showed that the local isolate B.licheniformis HM14 get higher yield of alpha – amylase production(102.10 unit/mg protein) according to killing percentage (90%) while the chemical mutation results showed that the local isolate B.licheniformis HM4 get higher yield of alpha –amylase production(100.94 unit/mg protein) from the two mutant local isolates (HM14 and HM4)were the best carbon source starch (1.5%), peptone (1.5%) as nitrogen source, calcium chloride (0.02%), sodium chloride (0.05%), magnicium phosphate (0.05%), sodium di –hydrogen phosphate (0.16%), at initial pH (5) and inoculum size 1*108 cfu/ml at (50?C) For (72) hours, using shaking incubator at (150) rpm.

Deciphering genome sequence of Paenibacillus illinoisensis strain YWY-3.1: A chitinase, cellulase, and amylase producer

The chitin-degrading Paenibacillus illinoisensis YWY-3.1 was isolated from the lake water at Yok Don National Park, Vietnam. Previous evaluations reported that strain YWY-3.1 had high activities of chitinases, cellulases, and amylases; it also produced remarkable levels of numerous promoting traits for plant growth. This study aimed to sequence and analyze the genome sequence of strain YWY-3.1 for further explorations and applications in agriculture and related fields. The whole genome sequencing revealed the P. illinoisensis YWY-3.1 genome contained 6,451,623 bp, 47.3 % GC, 5786 coding sequences, 88 tRNA, and 1 rRNA. It shared 95.49 % ANI and 87.1 % dDDH to those of P. illinoisensis NBRC 15959 (GCA 004000925). Among the coding sequences, COG deduced 4,991, and KEGG predicted 2819. The genome harbored 338 carbohydrate-active enzymes, including 195 GHs, 44 GTs, 13 PLs, 37 CEs, 3 AAs, and 46 CBMs; among them, 6 GH18 chitinases, 5 GH5 cellulases, 2 GH10 xylanases, 1 GH11 xylanase, and 2 GH13 α-amylases were identified. It contained 49 genes involved in environmental adaptation and plant growth. Moreover, 7 BGCs, with 3 being novel clusters, were identified from the genome. This work provided insight into the genomic information of P. illinoisensis YWY-3.1 and potential gene resources, especially chitinases, cellulases, xylanases, α-amylases, and novel BGCs for further explorations and applications.

Antimicrobial and Enzymatic Activities of Mangrove-associated Actinomycetes

This study delves into the enzymatic and antimicrobial capabilities of actinomycetes isolated from the Setiu Wetland mangrove in Terengganu, Malaysia. A total of eighteen actinomycete bacteria were isolated and characterized from the site. These isolates underwent antimicrobial assessments targeting a representative range of Gram-positive bacteria, Gram-negative bacteria and a fungus were employed for the testing. The results of the antimicrobial evaluations demonstrated pronounced effectiveness of the majority of isolated actinomycetes against Gram-negative bacterial strains. Intriguingly, a notable observation was the inhibition against Streptococcus uberis on nutrient agar by 27.7% of the isolates. In conjunction with the antimicrobial investigations, an array of enzymatic assays encompassing amylase, protease, lipase, phosphate solubilization, urease, and cellulase were executed. The outcomes revealed that a substantial portion of the examined actinomycetes exhibited positive reactions in at least half of the conducted assays, with amylase and protease production being particularly prominent, were observed from 94% of the isolates. These findings, drawn from the amassed dataset, underscore the remarkable diversity of antimicrobial and enzymatic activities within the actinomycetes thriving in the mangrove environment. This diversity exemplifies the adaptability of these mangrove-associated actinomycetes, underscoring their capacity to generate a versatile spectrum of secondary metabolites and biochemical responses as a strategy for survival within this unique ecosystem.

Amylase activity across black soldier fly larvae development and feeding substrates: insights on starch digestibility and external digestion

Black soldier fly larvae (BSFL; Hermetia illucens) hold promise for converting biowaste into proteins and lipids for feed. Dietary starch is efficiently digested by the larvae and influences larval performance, but the mechanisms of starch digestion remain poorly understood. This study investigated changes in individual weight and amylase activity in BSFL after 4, 7 and 11 days of feeding for five substrates varying in starch content and type: chicken feed (CF), corn gluten feed (CGF), wheat bran (WB), wheat distillers grain (WDG) and discarded potatoes (DP). Substrate amylase activities were also measured with and without larvae (feeding and fermenting trays, respectively) over time in order to explore external digestion. Feed conversion ratio (FCR) and estimated digestibility (ED) of DM and starch were assessed at the end of the experiment. The ranking for best FCR was CF, WB, CGF, WDG and DP. In feeding trays, ED of DM was 69.8 ± 1.8, 59.5 ± 2.9, 58.6 ± 0.7, 45.4 ± 0.6 and 19.5 ± 0.8% in CF, DP, WB, CGF and WDG, respectively. Estimated digestibility of starch reached 100% with WB and CGF, followed by CF (88.2 ± 2.3%), DP (85.2 ± 1.2%) and WDG (43.1 ± 1.0%). Larval amylase activity increased with growth for all substrates and dropped when approaching pupation. No relationship was found between larval amylase activity and substrate starch or other nutrient content, but a negative correlation was reported with the reducing sugar content of the larvae, suggesting glucose repression of amylase production. Amylase activity decreased with time in all feeding and fermenting substrates except WDG and DP. In vitro degradation assays indicated that BSFL amylase was nine times more efficient on raw corn or wheat starch than on raw potato starch, highlighting that starch structure is a major driver of digestibility. Western blot analysis revealed the presence of BSFL amylase in the feeding substrate, hinting at external digestion. Larval amylase was purified to identify its optimal pH (5.0–6.5) and temperature (70 °C). These results highlight that starch content is not a major driver of amylase activity in BSFL and suggest that other non-investigated factors could have had a crucial impact on the activity of larval digestive enzymes, such as microbial community of the substrate and presence of amylase inhibitors. This study also provides insights into the evolution of BSFL digestive activity during their development and the occurrence of external digestion.

Evaluation of the potential probiotic Bacillus subtilis isolated from darkbarbel catfish (Pelteobagrus fulvidraco) on growth performance, serum immunity, and disease resistance of Aeromonas hydrophila.

This study aimed to identify potential probiotic strains of Bacillus subtilis from healthy fish gut microbiota for application in aquaculture. The effects of dietary B. subtilis administration on growth performance, serum enzyme activity, immune gene expression, and disease resistance in darkbarbel catfish (Pelteobagrus fulvidraco) were investigated. The isolate, identified through gene sequencing and biochemical tests, demonstrated resilience to pH 3.0% and 6.0% bile, and exhibited extracellular protease, cellulose, lipase, and amylase production. Darkbarbel catfish were fed diets with varying B. subtilis concentrations (0 CFU/kg [T0], 107 CFU/kg [T1], 108 CFU/kg [T2], and 109 CFU/kg [T3]). After 8 weeks, significant increases (p < 0.05) were observed in final body weight, weight gain rate, specific growth rate, serum lysozyme, serum superoxide dismutase, alkaline phosphatase, and total antioxidant capacity, whereas malondialdehyde levels significantly decreased. Feeding darkbarbel catfish with B. subtilis diets increased immunoglobulin M (IgM) and C3 gene expression (p < 0.05), indicating a positive impact on the fish's immune system. The strain upregulated interleukin 10 (IL-10) and transforming growth factor-β (TGF-β) expression and downregulated TNF-α and IL-1β, suggesting potential anti-inflammatory effects. Following a 7-day challenge with Aeromonas hydrophila, fish fed with B. subtilis exhibited lower mortality, with higher survival rates in the T2 and T3 groups. In conclusion, supplementing darkbarbel catfish diets with B. subtilis effectively enhances growth performance, immune response, and disease resistance.

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.

Potential antimicrobial and fruit juice clarification activity of amylase enzyme from Bacillus strains

The hydrolytic enzyme, amylase possesses wide industrial applications and its production from bacterial sources by submerged fermentation is much simplified and economical. The research aimed to characterize amylase-producing bacteria and evaluate their potential for amylase activity regarding antimicrobial and fruit juice clarification. In current study, Bacillus licheniformis, Bacillus amyloliquifaciens, Bacillus cereus, Bacillus subtilis and Bacillus paramycoides was identified by 16S rRNA sequencing. After submerged fermentation, amylase activity of bacteria was measured by 3, 5-dinitro salicylic acid (DNS) assay. A substantial amount of amylase (423.47 mg/ml) in crude extract was measured by Bradford protein assay. Later, ammonium sulfate (80 %) precipitated partially purified amylase showed 1.6 times enhanced amylase activity (1484.94 U/ml) compared to crude amylase (973.23 U/ml). For highest amylase production, 72 h of optimum fermentation period was recorded at pH 7 with 2 % starch as substrate. Potent thermophilic amylase activity was observed at 65 °C. In apple juice clarification activity of amylase, turbidity of juice was reduced to 54.18 %. Potential antimicrobial property of amylase was detected with largest zone of inhibition against Escherichia coli ATCC 25922 (22.36 mm) and Mucor sp. ATCC 48559 (22.45 mm). Considering promising amylase properties, amylase-producing Bacillus strains from rice mill soil can be fermented for large scale amylase production providing application for industrial purposes including fruit juice clarification and antimicrobial activities. It will also overthrow the requirement of employing expensive and harmful chemicals in fruit juice clarification and combating pathogens.

Review of amylase production by microorganisms and their industrial application

Review of amylase production by microorganisms and their industrial application

Enhanced extracellular production of maltotetraose amylase from Pseudomonas saccharophila in Bacillus subtilis through regulatory element optimization

Maltotetraose amylase (Mta) catalyzes the hydrolysis of amylaceous polysaccharides into maltotetraose, which is an important functional sugar used in the food industry. However, the lack of efficient expression systems for recombinant Mta has hindered its scale-up production and application. In this study, a codon-optimized mta gene from Pseudomonas saccharophila was efficiently produced in Bacillus subtilis by optimizing the regulatory elements. First, a plasmid library containing 173 different signal peptide sequences placed upstream of mta gene was constructed, and transformed into B. subtilis strain WB800N(amyEΔ1) for high-throughput screening. The signal peptide yhcR was found to significantly enhance the secretion of Mta, reaching an activity of 75.4 U/mL in the culture medium. After optimization of the promoters, the Mta activity was further increased to 100.3 U/mL using a dual-promoter PHpaIIPamyE. Finally, the carbon sources and nitrogen sources for recombinant Mta production were optimized, yielding a highest Mta activity of 288.9 U/mL under the optimal culture conditions. The crude enzyme solution containing recombinant Mta produced a highest maltotetraose yield of 70.3% with 200 g/L of maltodextrin as the substrate. Therefore, the present study have demonstrated a high yield of Mta produced in B. subtilis, laying the foundation for large-scale Mta production and application.

Microbial Multienzyme Viz., Pectinase, Cellulase and Amylase Production Using Fruit and Vegetable Waste as Substrate—A Review

Around 70 million metric tonnes of fruit and vegetable waste (FVW) are produced each year and are eventually discarded as wholesale garbage. Microorganisms decompose this FVW, which has led to environmental contamination, greenhouse gas emissions, and other impacts related to climate change. If FVW are used properly, they can reduce environmental damage and also boost a nation’s economy. FVW contain vast amounts of biopolymers, viz., pectin, cellulose, and starch, all of which are hydrolysed by microbes with the aid of the pectinase, cellulase, and amylase enzymes, respectively. Therefore, in light of this, the intervention of microorganisms for the production of pectinase, cellulase, and amylase could be a safe, cost-effective, and eco-friendly approach for the precise utilisation of FVW. Nowadays, thermophilic multienzymes are extracted from a group of hot spring microbes. Thermophilic multienzymes are more capable of surviving at high temperatures and have less degrading capability. Moreover, through this advancement, we can obtain vast amounts of pectinase, cellulase, and amylase enzymes within a short period of time. This microbial enzyme preparation might be helpful in food, textiles, paper, pulp, animal feed supplements, detergents, juice/pulp clarity, leather, and other related sectors.

Phenotypic and Genotypic Characterization of Resistance and Virulence Markers in Candida spp. Isolated from Community-Acquired Infections in Bucharest, and the Impact of AgNPs on the Highly Resistant Isolates.

This study aimed to determine, at the phenotypic and molecular levels, resistance and virulence markers in Candida spp. isolated from community-acquired infections in Bucharest outpatients during 2021, and to demonstrate the efficiency of alternative solutions against them based on silver nanoparticles (AgNPs). A total of 62 Candida spp. strains were isolated from dermatomycoses and identified using chromogenic culture media and MALDI-TOF MS, and then investigated for their antimicrobial resistance and virulence markers (VMs), as well as for metabolic enzymes using enzymatic tests for the expression of soluble virulence factors, their biofilm formation and adherence capacity on HeLa cells, and PCR assays for the detection of virulence markers and the antimicrobial activity of alternative solutions based on AgNPs. Of the total of 62 strains, 45.16% were Candida parapsilosis; 29.03% Candida albicans; 9.67% Candida guilliermondii; 3.22% Candida lusitaniae, Candia pararugosa, and Candida tropicalis; and 1.66% Candida kefyr, Candida famata, Candida haemulonii, and Candida metapsilosis. Aesculin hydrolysis, caseinase, and amylase production were detected in the analyzed strains. The strains exhibited different indices of adherence to HeLa cells and were positive in decreasing frequency order for the LIP1, HWP1, and ALS1,3 genes (C. tropicalis/C. albicans). An inhibitory effect on microbial growth, adherence capacity, and on the production of virulence factors was obtained using AgNPs. The obtained results in C. albicans and Candida non-albicans circulating in Bucharest outpatients were characterized by moderate-to-high potential to produce VMs, necessitating epidemiological surveillance measures to minimize the chances of severe invasive infections.

A key component Rxt3 in the Rpd3L histone deacetylase complex regulates development, stress tolerance, amylase production and kojic acid synthesis in Aspergillus oryzae.

Rpd3L is a highly conserved histone deacetylase complex in eukaryotic cells and participates in various cellular processes. However, the roles of the Rpd3L component in filamentous fungi remain to be delineated ultimately. In this study, we constructed two knockout mutants of Rpd3L's Rxt3 subunit and characterized their biological functions in A. oryzae. Phenotypic analysis showed that AoRxt3 played a positive role in hyphal growth and conidia formation. Deletion of Aorxt3 resulted in augmented tolerance to multiple stresses, including cell wall stress, cell membrane stress, endoplasmic reticulum stress, osmotic stress and oxidative stress. Noteworthily, we found that Aorxt3-deleting mutants showed a higher kojic acid production than the control strain. However, the loss of Aorxt3 led to a significant decrease in amylase synthesis. Our findings lay the foundation for further exploring the role of other Rpd3L subunits and provide a new strategy to improve kojic acid production in A. oryzae.

Isolation of α-Amylase Producing Microorganisms from Soil of Kachchh, Gujarat

The purpose of this study is to explore the soil of the Gujarat, Kachchh region to identify amylase-producing bacteria and characterize them using molecular methods. The unique ecological characteristics of the Kachchh region may facilitate the isolation of these bacteria. Samples were collected from multiple locations within the Kachchh District, including Gandhidham, Rapar, Bhuj, Nakhatrana, Mandvi, and Mundra Talukas. These samples were then screened to isolate amylase-producing bacteria. A total of 27 different types of colonies were identified, out of which 16 exhibited amylase production (M1-M16). Out of 27 colonies identified, 16 showed amylase production. Strains M2, M7, and M13 exhibited high amylase activity, with M2 showing a consistent increase over 72 hours, making it a strong candidate for amylase production. Further identification of M2 stain identified M2 as a Gram-positive, spore-forming, capsulated, and motile bacillus, specifically Bacillus licheniformis. This was confirmed through DNA sequencing and analysis in the NCBI database, which showed a 99.15% similarity with Bacillus licheniformis. The study concludes that Soil in Kachchh is rich with microorganisms that produce amylase, an enzyme with diverse industrial applications. These organisms are valuable for sectors like food, textiles, paper, detergents, pharmaceuticals, and biofuel production.

Influence of enzyme drug and adsorbent on digestive metabolism processes of steers fattening in technogenic zone

In the practice of feeding of young ruminants for fattening, feed drugs of new generation adsorbents are increasingly and widely introduced. They often exhibit synergistic eff ects on metabolic processes with a large set of biologically active additives including enzyme drugs. The purpose of the research was to study the infl uence of the adsorbent drugs toxfi n and MEC celloviridin G20x on the state of rumen metabolism in fattening steers in whose diets there was an excess content of lead, zinc and cadmium salts. It was found that in order to activate the processes of digestive metabolism due to better elimination of heavy metal salts, it is advisable to jointly introduce the adsorbent toxfi n in the amount of 1 kg/t of compound feed and MEC celloviridin G20x in the amount of 70 g/t of compound feed into the composition of the rations of fattening steers. The results of the conducted studies showed that by the end of fattening at the age of 18 months the animals of the 4th experimental group had the highest body weight, which surpassed the steers of the 1st control group by 29.20 kg (p &lt; 0.05) or 6.86 %. Relative to the control analogues the steers of the 4th experimental group showed an activation of growth in the forestomachs of ciliates (producers of cellulases and amylases) by 54.26 % (p &lt; 0.05) and bacteria of the Flavobacterium vitarumen group (producers of proteinases) by 29.36 % (p &lt; 0.05). Due to the combined feeding of the adsorbent drugs toxfi n and MEC celloviridin G20x, the processes of detoxifi cation of heavy metal salts were more effective. Therefore, the fattening steers of the 4th experimental group more actively accumulated volatile fatty acids in the rumen fl uid, significantly (p &lt; 0.05) outperforming the animals of the 1st control group in this indicator by 1.68 mmol/100 ml or by 20.71 %.