Maximum Enzyme Production Research Articles

Screening and Optimization of Pathogenic Strains of Pseudomonas sp. for Cholinesterase Enzyme Activity

The identification and optimization of pathogenic strains of Pseudomonas sp. with significant Cholinesterase enzyme activity has promising implications for environmental and medical applications. This study aimed to screen various Pseudomonas strains to identify those exhibiting high cholinesterase activity and to optimize the conditions for maximal enzyme production. Multiple Pseudomonas strains were isolated and characterized from diverse clinical samples, employing selective media and biochemical assays to determine cholinesterase activity. The strains with maximum enzyme activity were further subjected to optimization experiments, including variations in growth conditions such as Temperature, pH and Nutrient availability. The findings revealed sample no.39 among the collected 50 samples had significant enzyme activity of 0.19151 IU is obtained when choline chloride was 0.3999%, Ammonium Chloride was 0.01M, pH was 7.0, Temperature was 27°C and Time of incubation was 24hrs. These results not only advance our understanding of Pseudomonas sp. enzyme capabilities but also offer potential applications in bioremediation, biosensor development and pharmaceutical industries. Future research will focus on the molecular characterization of the cholinesterase enzyme and its applications in various industrial processes.

Kinetics of cellulase-free endo xylanase hyper-synthesis by Aspergillus Niger using wheat bran as a potential solid substrate

The present study deals with the production of cellulase-free endoxylanase by Aspergillus niger ISL-9 using wheat bran as a solid substrate. Endoxylanase was produced under a solid-state fermentation. Various growth parameters were optimized for the improved production of the enzyme. The Substrate level of 15 g was optimized as it provided the fungus with balanced aeration and nutrition. Among the six moisture contents investigated, Moisture Content 5 (MC5) was optimized (g/l: malt extract, 10; (NH4)2HPO4, 2.5; urea, 1.0) and 10 mL of MC5 was found to give the highest production of endoxylanase. The pH and time of incubation were optimized to 6.2 and 48 h respectively. The Inoculum size of 2 mL (1.4 × 106 spores/mL) gave the maximum enzyme production. After optimization of these growth parameters, a significantly high endoxylanase activity of 21.87 U/g was achieved. Very negligible Carboxymethylcellulase (CMCase) activity was observed indicating the production of cellulase-free endoxylanase. The notable finding is that the endoxylanase activity was increased by 1.4-fold under optimized conditions (p ≤ 0.05). The overall comparison of kinetic parameters for enhanced production of endoxylanase by A. niger ISL-9 under Solid State Fermentation (SSF) was also studied. Different kinetic variables which included specific growth rate, product yield coefficients, volumetric rates and specific rates were observed at 48, 72 and 96 h incubation time and were compared for MC1 and MC5. Among the kinetic parameters, the most significant result was obtained with volumetric rate constant for product formation (Qp) that was found to be optimum (1.89 U/h) at 72 h incubation period and a high value of Qp i.e.1.68 U/h was also observed at 48 h incubation period. Thus, the study demonstrates a cost-effective and environmentally sustainable process for xylanase production and exhibits scope towards successful industrial applications.

Production of milk-coagulating protease by fungus Pleurotus djamor through solid state fermentation using wheat bran as the low-cost substrate

Proteases are enzymes that hydrolyze peptide bonds present in proteins and peptides. They are widely used for various industrial applications, such as in the detergent, food, and dairy industries. Cheese is one of the most important products of the dairy industry, and the coagulation stage is crucial during the cheese-making process. Enzymatic coagulation is the most common technique utilized for this purpose. Microbial enzymes are frequently used for coagulation due to their advantages in terms of availability, sustainability, quality control, product variety, and compliance with dietary and cultural/religious requirements. In the present study, we identified and subsequently characterized milk coagulant activity from the fungus Pleurotus djamor PLO13, obtained during a solid-state fermentation process, using the agro-industrial residue, wheat bran, as the fermentation medium. Maximum enzyme production and caseinolytic activity occurred 120 h after cultivation. When the enzyme activity against various protease-specific synthetic substrates and inhibitors was analyzed, the enzyme was found to be a serine protease, similar to elastase 2. This elastase-2-like serine protease was able to coagulate pasteurized whole and reconstituted skim milk highly efficiently in the presence and absence of calcium, even at room temperature. The coagulation process was influenced by factors such as temperature, time, and calcium concentration. We demonstrate here, for the first time, an elastase-2-like enzyme in a microorganism and its potential application in the food industry for cheese production.

Optimizing bioethanol production from hassawi rice straw with Aspergillus sp. NAS51 cellulosic enzyme and in silico homology modeling

Researchers' attention has been greatly focused on the use of lignocellulosic wastes to produce bioethanol and biogas due to the depletion of non-renewable energy resources. In our effort to find a potent cellulase-producing fungal strain, the fungus NAS51 was isolated among eight isolates from a sponge collected from the Red Sea, Jeddah, and selected as it displayed the highest cellulase activity including (FP-ase, CMC-ase, and β-glucosidase enzymes) at levels of 3.13 U/ml, 2.52 U/ml, and 0.69 U/ml, respectively. The fungus was identified morphologically and genetically by sequencing its 18SrRNA gene as Aspergillus sp. NAS51. The cellulase activity of Aspergillus sp. NAS51 was optimized and maximum enzyme production was obtained at initial pH7, temp 30oC, incubation period 11 days, moisture content 70%, urea as a nitrogen source, and K2HPO4 (2 g/L). The crude cellulases from strain NAS51 were characterized, and results showed that the enzyme is stable over a wide pH range (6–10), with peak activity at pH 7.0 and 40 °C. The cellulase gene has been sequenced and the protein 3D structure was generated via in silico homology modeling. Determination of binding sites and biological annotations of the constructed protein was carried out via COACH and COFACTOR based on the I-TASSER structure prediction. To reach the maximum enzyme hydrolysis, the rice straw collected from Al-Ahsa, Kingdom of Saudi Arabia was pretreated with NaOH 1.5% to remove lignin and to enhance the saccharification process by Cellulase. The saccharified product was measured using HPLC, fermented by S. cerevisiae and the bioethanol yield produced from the fermentation was 0.454 mL ethanol/g fermentable sugars. This study indicates the potential application of fungal enzymes such as cellulase enzymes in biofuel generation and waste management.

Mechanisms of Cell Wall Degrading Enzymes from Bacillus methylotrophicus and Bacillus subtilis in Suppressing Foliar Blight Pathogens

Bacillus strains are potent Biocontrol agents (BCAs) and have been identified as an effective way to control the growth of phytopathogens of wheat. Through a direct inhibition mechanism which involves the production of cell wall degrading enzymes (proteases, glucanase, and chitinases) and siderophores they suppress the foliar blight disease. Both the strains of Bacillus P10 and UP11 shows growth inhibition Alternaria triticina (77.56%, 67.83%) and Bipolaris sorokiniana (73.97%, 62.16%) through a dual culture assay. These strains were subsequently examined for their ability to produce cell wall-degrading enzymes i.e., chitinase, protease, and β-1,3-glucanases and antifungal metabolites i.e., siderophores. It was found that when the antagonist’s bacteria were Co-cultured with fungal pathogens then maximum production of hydrolytic enzymes and siderophore was achieved at 96 hrs, but when both strains P10 and UP11 were alone maximum production was found at 48 hrs i.e., the exponential phase. The current study determined that Bacillus methylotrophicus (P10) and Bacillus subtilis (UP11) are highly effective strain for controlling Foliar Blight disease. Direct or Antibiosis is the main mechanism involved in this study. Further research on the interaction mechanisms between Bacillus-derived compounds and host plants is necessary.

Optimization of newly isolated Bacillus cereus α-amylase production using orange peels and crab shells and application in wastewater treatment.

A newly isolated amylolytic strain was identified as Bacillus cereus spH1 based on 16S and 16-23S gene sequencing (Accession numbers OP811441.1 and OP819558, respectively), optimization strategies, using one variable at time (OVAT) and Plackett-Burman design, were employed to improve the alpha-amylase (α-amylase) production. Condition inferred revealed that the optimal physical parameters for maximum enzyme production were 30°C, pH 7.5, and 12h of incubation, using tryptone, malt extract, orange (Citrus sinensis) peels, crab (Portunus segnis) shells, calcium, and sodium chloride (NaCl) as culture medium. The full factorial design (FFD) model was observed to possess a predicted R2 and adjusted R2 values of 0.9788 and 0.9862, respectively, and it can effectively predict the response variables (p = 0). Following such efforts, α-amylase activity was increased 141.6-folds, ranging from 0.06 to 8.5 U/mL. The ideal temperature and pH for the crude enzyme activity were 65°C and 7.5, respectively. The enzyme exhibited significant stability, with residual activity over 90% at 55°C. The maltose was the only product generated during the starch hydrolysis. Moreover, the Bacillus cereus spH1 strain and its α-amylase were used in the treatment of effluents from the pasta industry. Germination index percentages of 143% and 139% were achieved when using the treated effluent with α-amylase and the strain, respectively. This work proposes the valorization of agro-industrial residues to improve enzyme production and to develop a green and sustainable approach that holds great promise for environmental and economic challenges.

Statistical optimization of cellulase production from Bacillus sp. YE16 isolated from yak dung of the Sikkim Himalayas for its application in bioethanol production using pretreated sugarcane bagasse

Cellulolytic bacteria were isolated from yak dung samples collected from different habitats of Sikkim, India. Isolate YE16 from the Yumthang Valley sample showed highest enzyme activity of 7.68 U/mL and was identified as Bacillus sp., which has a sequence similarity of 96.15% with B. velezensis. One factor at a time (OFAT) analysis revealed that an acidic pH of 5 with 37 °C temperature was optimum for maximum enzyme production after 36 hrs of incubation (13.88 U/mL), which was further increased after statistical optimization (34.70 U/mL). Media optimization based on response surface methodology predicted that Carboxymethyl cellulose (CMC) and MgSO4 at concentrations of 30 g/L and 0.525 g/L, respectively, at pH 5.5 to show CMCase activity of 30.612 U/mL, which was consistent with the observed value of 30.25 U/mL and confirmed the model. The crude enzyme also efficiently hydrolyzed alkaline pretreated sugarcane bagasse, releasing 7.09 g/L of glucose equivalent with an ethanol production of 3.05 g.

Sustainable production and preparative purification of thermostable alkaline α-amylase by Bacillus simplex (ON754233) employing natural deep eutectic solvent-based extractive fermentation

Using PEG-based deep eutectic solvents (PDES), the current study proposes extractive fermentation as a sustainable process integration for the production and purification of α-amylase from Bacillus simplex (ON754233). Glucose: PEG 400 outperformed five PDES in terms of tie lie length (58) and slope value (1.23) against sodium sulphatt. Apple cider pomace was used as a low-cost, sustainable carbon source to produce-amylase, with a maximum enzyme production of 2200.13 U/mL. PDES concentration (20% w/v), salt (12.75 w/v), and apple waste (2.75 g/mL) were all optimized using response surface methodology. When scaled upto 3 L benchtop bioreactor, extractive fermentation was proved to be better technology with maximum recovery of 92.4% with highest partition coefficient (3.59). The partially purified enzyme was further purified using a Sephadex G 100 followed by DEAE-Sephadex anion exchange chromatography with a purity fold of 33. The enzyme was found to be thermostable at the temperature (60 °C), remains alkaline (pH 8), and the activity was stimulated in the presence of Mg2+ ions. With SDS PAGE electrophoresis, the molecular weight was found to be around 140 kDa. Finally, the enzyme kinetics parameters were evaluated with observed Km (0.00396 mM) and Vmax (37.87 U/mL). Thus scaling up extractive fermentation entails increasing production capacity with improved extraction efficiency using green solvents.

Immobilization and biochemical characterization of purified L-Asparaginase produced by Brevibacillus borstelensis ML12

Purified L-Asparaginase from Brevibacillus borstelensis ML12 was immobilized, and sequentially studied for biochemical characterization and thermodynamic feasibility. Immobilization has long been used as a way to enhance enzyme stability, facilitate recovery, and multiple re-uses in industries. It is preferable to utilize natural nontoxic carriers and crosslinking agents to immobilize enzymes. L-Asparaginase is an important industrial enzyme due to its significant role in the pharmaceutical and food sectors. In this study, immobilization of purified L-Asparaginase from a novel bacterium Brevibacillus borstelensis ML12 was performed using different carrier matrices and concentration of the suitable matrix was varied to obtain maximum enzyme production. Biochemical parameters like effect of pH, temperature, surfactants, organic reagents, bile salt were also studied. Arrhenius plot and thermal stability of immobilized enzyme were obtained from thermodynamic studies. ΔH, ΔG and ΔS values were also determined. Operational and storage stability studies revealed that the enzyme was able to retain 80% of its activity after 3 cycles and relative activity falls by 45% after 48 hours under refrigerated conditions and by 50% at normal room temperature. Vmax and KM values were calculated by using Lineweaver-Burk plot and the values came out to be 121.654 µmole ml-1 min-1 and 0.310 mM respectively.

Biosoftening of banana pseudostem fiber using cellulase and pectinase enzyme isolated from Aspergillus niger for textile industry

BackgroundNowadays, farmers are facing a lot of problems for the disposal of banana pseudostem waste after the harvesting of banana. Banana pseudostem is a rich source of fiber, which is an alternative source of other natural and synthetic fibers. The banana fibers are biodegradable, and they are expected to be in great demand in the international market. For the textile industry, fibers were extracted using chemical and mechanical methods, but it leads to damage and affects the quality of fibers. So, this study mainly focused on biosoftening of banana pseudostem fiber using crude enzyme produced from Aspergillus niger which is one of the most predominant fungus which can synthesize industrially applicable enzymes and which can soften the surface of banana pseudostem fiber. Through this, biosoftened banana pseudostem fiber can be produced, and the disposal problem of banana pseudostem can be rectified in an eco-friendly manner. ResultThe present study was undertaken for the biosoftening of banana pseudostem fiber using crude enzymes isolated from fungal strain. The fungal isolates were subjected to enzyme screening such as cellulase, pectinase, chitinase, peroxidase, and polygalacturonase. The maximum production of enzyme was observed in F2 strain, and it was subjected to crude enzyme production and purification using dialysis and column chromatography. The collected best enzyme fractions were selected for the biosoftening of banana pseudostem fiber. The banana pseudostem fiber was treated with crude enzymes at the time duration of 2, 4, 24, and 48 h. After the treatment, the treated and untreated fibers were evaluated for the mechanical properties and chemical constituent’s analysis. The results revealed that the chemical contents were high during 2- and 4-h-treated fibers. After that, chemical constituents were reduced due to the removal of debris by the action of enzymes. The mechanical properties such as breaking load, breaking extension, tenacity, and diameter of fiber were best in the fibers treated for 2 and 4 h. After 4 h due to the removal of chemical constituents the breaking load and tenacity, diameter will be reduced. SEM results proved that the fiber treated at 4th h showed smooth and softened fiber. ConclusionThis study proved that the crude enzymes isolated from the Aspergillus niger can be effectively soften and increase the quality of banana pseudostem fiber.

Optimization of tannase production by Aspergillus glaucus in solid-state fermentation of black tea waste

Tannases are valuable industrial enzymes used in food, pharmaceutical, cosmetic, leather manufacture and in environmental biotechnology. In this study, 15 fungal isolates were obtained from Egyptian cultivated soil and marine samples. The isolated fungi were qualitatively and quantitatively screened for their abilities to produce tannase. The selected fungal isolate NRC8 giving highest tannase activity was identified by molecular technique (18S rRNA) as Aspergillus glaucus. Among different tannin-containing wastes tested, the black tea waste was the best substrate for tannase production by Aspergillus glaucus in solid-state fermentation (SSF). Optimization of the different process parameters required for maximum enzyme production was carried out to design a suitable SSF process. Maximal tannase production was achieved with moisture content of 75%, an inoculums size of 6 × 108 spore/ml and sodium nitrate 0.2% (pH of 5.0) at 30 °C after 5 days of incubation. Box–Behnken experiment was designed to get a quadratic model for further optimization studies. Four-factor response-surface method with 27 runs was prepared using independent parameters including (moisture content %, initial pH, substrate concentration (g) and sodium nitrate concentration (g) for tannase model. The F- and P-values of the model were 4.30 and 0.002, respectively, which implied that the model is significant. In addition, the lack-of-fit was 1040.37 which indicates the same significance relative to the pure error. A. glaucus tannase was evaluated by the efficiency of conversion of tannic acid to gallic acid. Moreover, production of gallic acid from SSF process of A. glaucus using black tea waste was found to be 38.27 mg/ml. The best bioconversion efficiency was achieved at 40 °C with tannic acid concentration up to 200 g/L.Graphical

Temperature and pH Optimization for Protease Production Fermented by Yarrowia lipolytica from Agro-Industrial Waste

The use of yeasts for the production of proteases has increased in demand in recent years. Y. lipolytica has been reported as a strain with high yields of protease production. This work aimed to evaluate the impact of pH and temperature on the production of proteases using Y. lipolytica in solid-state fermentation (SSF). Soybean, canola meal, cottonseed meal, and sesame meal wastes were used as nutrient sources at seven pH levels (4, 5, 6, 7, 8, 9, 10) and five temperatures (25, 30, 35, 40, 45 °C). The waste source and optimal conditions for maximum enzyme production (EP) were obtained by Box–Benhken design. The results revealed that at pH of 7, temperature of 30 °C, and for 48 h cultivation period, canola meal showed the best EP with 188.75 U/L, followed by soybean with 117.07 U/L, cottonseed meal with 66.71 U/L, and sesame with the lowest production, reaching 88.5 U/L up to 35 °C. The temperature factor exhibited the greatest effect on protease production. The biotechnological and economic potential of canola meal in the production of enzymes is highlighted.

Valorization of lignocellulosic wastes for sustainable xylanase production from locally isolated Bacillus subtilis exploited for xylooligosaccharides’ production with potential antimicrobial activity

The worldwide availability of lignocellulosic wastes represents a serious environmental challenge with potential opportunities. Xylanases are crucial in lignocellulosic bio-hydrolysis, but the low enzyme productivity and stability are still challenges. In the current study, Bacillus subtilis (coded ARSE2) revealed potent xylanase activity among other local isolates. The enzyme production optimization revealed that maximum enzyme production (490.58 U/mL) was achieved with 1% xylan, 1.4% peptone, and 5% NaCl at 30 °C and pH 9. Furthermore, several lignocellulosic wastes were exploited for sustainable xylanase production, where sugarcane bagasse (16%) under solid-state fermentation and woody sawdust (2%) under submerged fermentation supported the maximum enzyme titer of about 472.03 and 485.7 U/mL, respectively. The partially purified enzyme revealed two protein bands at 42 and 30 kDa. The partially purified enzyme revealed remarkable enzyme activity and stability at 50–60 °C and pH 8–9. The enzyme also revealed significant stability toward tween-80, urea, DTT, and EDTA with Vmax and Km values of 1481.5 U/mL and 0.187 mM, respectively. Additionally, the purified xylanase was applied for xylooligosaccharides production, which revealed significant antimicrobial activity toward Staphylococcus aureus with lower activity against Escherichia coli. Hence, the locally isolated Bacillus subtilis ARSE2 could fulfill the xylanase production requirements in terms of economic production at a high titer with promising enzyme characteristics. Additionally, the resultant xylooligosaccharides revealed a promising antimicrobial potential, which paves the way for other medical applications.

Unleashing the Potential of Bacterial Isolates from Apple Tree Rhizosphere for Biocontrol of Monilinia laxa: A Promising Approach for Combatting Brown Rot Disease.

Monilinia laxa, a notorious fungal pathogen responsible for the devastating brown rot disease afflicting apples, wreaks havoc in both orchards and storage facilities, precipitating substantial economic losses. Currently, chemical methods represent the primary means of controlling this pathogen in warehouses. However, this study sought to explore an alternative approach by harnessing the biocontrol potential of bacterial isolates against brown rot in apple trees. A total of 72 bacterial isolates were successfully obtained from the apple tree rhizosphere and subjected to initial screening via co-cultivation with the pathogen. Notably, eight bacterial isolates demonstrated remarkable efficacy, reducing the mycelial growth of the pathogen from 68.75 to 9.25%. These isolates were subsequently characterized based on phenotypic traits, biochemical properties, and 16S rRNA gene amplification. Furthermore, we investigated these isolates' production capacity with respect to two enzymes, namely, protease and chitinase, and evaluated their efficacy in disease control. Through phenotypic, biochemical, and 16S rRNA gene-sequencing analyses, the bacterial isolates were identified as Serratia marcescens, Bacillus cereus, Bacillus sp., Staphylococcus succinus, and Pseudomonas baetica. In dual culture assays incorporating M. laxa, S. marcescens and S. succinus exhibited the most potent degree of mycelial growth inhibition, achieving 68.75 and 9.25% reductions, respectively. All the bacterial isolates displayed significant chitinase and protease activities. Quantitative assessment of chitinase activity revealed the highest levels in strains AP5 and AP13, with values of 1.47 and 1.36 U/mL, respectively. Similarly, AP13 and AP6 exhibited the highest protease activity, with maximal enzyme production levels reaching 1.3 and 1.2 U/mL, respectively. In apple disease control assays, S. marcescens and S. succinus strains exhibited disease severity values of 12.34% and 61.66% (DS), respectively, highlighting their contrasting efficacy in mitigating disease infecting apple fruits. These findings underscore the immense potential of the selected bacterial strains with regard to serving as biocontrol agents for combatting brown rot disease in apple trees, thus paving the way for sustainable and eco-friendly alternatives to chemical interventions.

Modeling and in-vivo evaluation of fibrinolytic enzyme produced by Bacillus subtilis Egy under solid state fermentation

Blood clot formation increases cases of myocardial infarction (AMI) and stroke, thus urges directing much research works for treatment and prevention of the causes. One of these directions is the microbial production of fibrinolytic enzymes as thrombolytic agents. In the current work, Bacillus subtilis Egy has been used for enzyme production under solid state fermentation. Among twelve nutrient meals in addition to wheat bran as a control fodder yeast yielded the highest enzyme activity reaching 114U/g. Applying statistical model for optimization of enzyme production revealed that 3.6%, fodder yeast; 40%, moisture content; 6 days, incubation period and 2%, inoculum size were the optimum conditions for maximum fibrinolytic enzyme production (141.02 U/g) by Bacillus subtilis Egy under solid-state fermentation The model was significant and data were experimentally validated. The produced fibrinolytic enzyme was evaluated for in vitro and in vivo cytotoxicity. In-vivo examination of the enzyme resulted in no mortality during the first 24 h after treatment. After 14 days, the results revealed no significant changes detected in hematological parameters (RBCs, MCV, hemoglobin except WBCs which showed an increase for both sexes. Histopathological examination of liver and kidney of rats received oral and subcutaneous treatments showed normal architecture. The data showed the applicability of the produced enzyme for the treatment of blood clot with no significant effect on living cells or on physiological functions.

Production, characterization, and application of a new chymotrypsin-like protease from Pycnoporus sanguineus

Proteases are the most commercialized enzymes and are widely used in industrial processes, especially in the dairy industry. Cheese production is increasing disproportionately to the availability of calf rennet, the main coagulant used in this process. In addition to the cost of rennet, there is also a religious and dietary objection to the use of calf proteases. An alternative is the production of microbial enzymes via solid state fermentation (SSF), which uses agroindustrial residues as a source of nutrients and substrate. To obtain proteases with milk-clotting activity, Pycnoporus sanguineus was produced via SSF using agroindustrial residue wheat bran as the culture medium. Caseinolytic activity and maximum enzyme production were obtained after 96 h of cultivation. The protease was identified as a chymotrypsin-like serine protease after testing it against specific substrates and inhibitors. The protease from P. sanguineus had a specific activity of 741.3 U mg−1 and was able to coagulate reconstituted skim milk and whole milk, with and without the addition of calcium. Coagulation was affected by time, temperature, and calcium and enzyme concentrations. Therefore, we describe herein for the first time a protease obtained from P. sanguineus, a chymotrypsin-like serine protease with milk-clotting activity that has potential applications in the dairy industry.

Rice straw derived graphene-silica based nanocomposite and its application in improved co-fermentative microbial enzyme production and functional stability

Cellulases are the one of the most highly demanded industrial biocatalysts due to their versatile applications, such as in the biorefinery industry. However, relatively poor efficiency and high production costs are included as the key industrial constraints that hinder enzyme production and utilization at economic scale. Furthermore, the production and functional efficiency of the β-glucosidase (BGL) enzyme is usually found to be relatively low among the cellulase cocktail produced. Thus, the current study focuses on fungi-mediated improvement of BGL enzyme in the presence of a rice straw-derived graphene-silica-based nanocomposite (GSNCs), which has been characterized using various techniques to analyze its physicochemical properties. Under optimized conditions of solid-state fermentation (SSF), co-fermentation using co-cultured cellulolytic enzyme has been done, and maximum enzyme production of 42 IU/gds FP, 142 IU/gds BGL, and 103 IU/gds EG have been achieved at a 5 mg concentration of GSNCs. Moreover, at a 2.5 mg concentration of nanocatalyst, the BGL enzyme showed its thermal stability at 60°C and 70 °C by holding its half-life relative activity for 7 h, while the same enzyme demonstrated pH stability at pH 8.0 and 9.0 for the 10 h. This thermoalkali BGL enzyme might be useful for the long-term bioconversion of cellulosic biomass into sugar.

L-Asparaginase – A prospective anticancer enzyme on MCF-7 cell lines extracted from soil Microflora

This study aims at investigating the soil bacteria for L asparaginase enzyme production and its anti-cancer effect on MCF 7 cell line. In this study, 30 soil samples were screened for bacterial producing L asparaginase enzyme. The soil samples were collected from various places in Ethiraj College, Chennai. Isolation was done using spread plate technique, biochemical and microscopic characterization for the potential isolates revealed the bacterial genera as Bacillus, Escherichia and Pseudomonas species. Of the thirty samples collected, six isolates produced pronounced L asparaginase enzyme. These samples were further subjected to crude enzyme extraction and estimation. The isolates S2 and Ni5 showed maximum production of enzyme with 1090 units/ml of activity. Dialysis was carried out to concentrate the enzyme and molecular characterization was performed using SDS PAGE to separate the proteins according to the molecular weight which was found to be approximately 100 to 116 kDa. Anti-cancer activity was evaluated on MCF 7 tumour cell line. The cell viability was dose dependent and IC50 of S2 was 62.5 μg and Ni5 31.2 μg respectively.

OPTIMIZATION OF PHYSICAL PARAMETERS FOR PRODUCTION OF AMYLASES AND PROTEASES FROM SOIL BACTERIA

Enzymes are biomolecules present in the cells of living organisms in minute amounts and are capable of speeding up chemical reactions, without being altered and utilized in the reaction. Microbial enzymes are preferred than both plant and animal sources because they are cost effective with respect to production, more predictable, controllable and reliable. These naturally occurring enzymes are quite often not readily available in sufficient quantities for food applications or industrial use. However, by isolating microbial strains that produce the desired enzyme and optimizing the conditions for growth, commercial quantities can be obtained. Our study aimed at isolating and identifying the bacterial strains from soil which produce extracellular proteases and amylases and the optimization of physical conditions for maximum enzyme production.

The potential of degrading natural chitinous wastes to oligosaccharides by chitinolytic enzymes from two Talaromyces sp. isolated from rotten insects (Hermetia illucens) under solid state fermentation.

It is difficult to produce chitin oligosaccharides by hydrolyzing untreated natural chitinous waste directly. In this study, two fungi Talaromyces allahabadensis Hi-4 and Talaromyces funiculosus Hi-5 from rotten black soldier fly were isolated and identified through multigene phylogenetic and morphological analyses. The chitinolytic enzymes were produced by solid state fermentation, and the growth conditions were optimized by combining single-factor and central composite design. The best carbon sources were powder of molting of mealworms (MMP) and there was no need for additional nitrogen sources in two fungi, then the maximum chitinolytic enzyme production of 46.80 ± 3.30 (Hi-4) and 55.07 ± 2.48 (Hi-5) U/gds were achieved after analyzing the 3D response surface plots. Pure chitin (colloidal chitin) and natural chitinous substrates (represented by MMP) were used to optimize degradation abilities by crude enzymes obtained from the two fungi. The optimum temperature for hydrolyzing MMP (40°C both in two fungi) were lower and closer to room temperature than colloidal chitin (55°C for Hi-4 and 45°C for Hi-5). Then colloidal chitin, MMP and the powder of shrimp shells (SSP) were used for analyzing the products after 5-day degradation. The amounts of chitin oligosaccharides from SSP and MMP were about 1/6 (Hi-4), 1/17 (Hi-5) and 1/8 (Hi-4), 1/10 (Hi-5), respectively, in comparison to colloidal chitin. The main components of the products were GlcNAc for colloidal chitin, (GlcNAc)2 for MMP, and oligosaccharides with higher degree of polymerization (4-6) were obtained when hydrolyzing SSP, which is significant for applications in medicine and health products.