Optimum Temperature For Enzyme Production Research Articles

Obtaining L - Glutamate Oxidase Enzyme from Streptomyces and Estimation Its Toxicity on Blood Components

Streptomyces was isolated from the soil and then tested to diagnose morphologically and microscopically. Also, cultivated in the production medium of the L-glutamate oxidase enzyme, for which the wheat bran culture media was used, and then the extracellular L-glutamate oxidase enzyme was extracted using a cooling centrifuge at 8000 rpm for 20 minutes. The optimal temperature and pH for the production of the enzyme’s conditions were studied. Research results revealed that the optimum temperature for enzyme production was 30°C, and the best pH for the enzyme’s production was 7.0 later on, the enzyme was purified using common purification techniques represented by precipitation with ammonium sulfate as a preliminary purification step, followed by dialysis to remove salts, ion-exchange chromatography and gel filtration chromatography were used to finish purification process. The effect of the enzyme toxicity on the human blood components was evaluated. The enzyme had no harmful effect on the blood cells, with Red Blood Cells reaching 4 x 1012, White Blood Cells 7.5 × 109 and Platelets 293 × 109.

Molecular Characterization of Trichoderma asperellum and Lignocellulolytic Activity on Barley Straw Treated with Silver Nanoparticles

Silver nanoparticles (AgNPs) have been applied as an antifungal agent, which results in AgNPs contamination of agricultural waste that interferes with the lignocellulosic enzymes produced by fungi. Therefore, this study examined the production of carboxymethylcellulase (CMCase) and manganese-dependent lignin peroxidase (MnPase), using barley straw treated with AgNPs. Trichoderma asperellum growth was not inhibited at 25 ppm AgNPs, while negligible growth inhibition was observed at 50 ppm AgNPs, which was not observed with Aspergillus terreus and Curvularia lunata. T. asperellum was the highest producer of CMCase and MnPase using barley straw with or without 25 ppm AgNPs versus A. terreus or C. lunata. AgNPs addition to barley straw before T. asperellum inoculation played a role in repressing enzyme activities (CMCase 156.33 U/mL and MnPase 1.28 U/mL); however, addition of AgNPs (50 ppm) after 10 days of incubation showed the highest activity (CMCase 160.67 and 1.35 U/mL MnPase). Optimum temperature for enzyme production by T. asperellum using untreated and treated barley straw was 35 °C and 30 °C, respectively. Enzyme activities increased with increasing polyoxyethylenesorbitan monooleate surfactant concentrations up to 0.25 mL/g substrate without AgNPs, whereas the activity decreased with AgNPs (25 ppm). The exception to this observed trend was at low concentrations of the surfactant (0.10 mL/g substrate).

Studies on the production of glucose isomerase by Bacillus licheniformis

Abstract This work reports the effects of some culture conditions on the production of glucose isomerase by Bacillus licheniformis. The bacterium was selected based on the release of 3.62 mg/mL fructose from the fermentation of glucose. Enzyme was produced using a variety of carbon substrates but the highest enzyme activity was detected in a medium containing 0.5% xylose and 1% glycerol (specific activity = 6.88 U/mg protein). Media containing only xylose or glucose gave lower enzyme productivies (specific activities= 4.60 and 2.35 U/mg protein respectively). The effects of nitrogen substrates on glucose isomerase production showed that yeast extract supported maximum enzyme activity (specific activity = 5.24 U/mg protein). Lowest enzyme activity was observed with sodium trioxonitrate (specific activity = 2.44 U/mg protein). In general, organic nitrogen substrates supported higher enzyme productivity than inorganic nitrogen substrates. Best enzyme activity was observed in the presence of Mg2+ (specific activity = 6.85 U/mg protein) while Hg2+ was inhibitory (specific activity = 1.02 U/mg protein). The optimum pH for best enzyme activity was 6.0 while optimum temperature for enzyme production was 50ºC.

دراسة البکتریا المنتجة لانزیم السلیولیز المعزولة من بیئات مختلفة وایجاد الظروف المثلى لانتاج الانزیم.

One hundred eighty samples were collected from (soil, insect intestine, leaves, stomach of ruminants, and sewage water) and only five isolates were selected depend on its ability to lyses cellulose and they were one from each isolation source , the selected isolates were belonged to (Staphylococcus sciuri , Streptococcus parasanguinis , Sphingomonaspaucimobilis , Rhizobium radiobacter , Escherichia coli ) with lyses zone (42,28,39,35,32 mm respectively). The PH effect change showed that best activity for cellulase enzyme was 6.627 IU/ml at PH 6 when local isolate Staph. Sciuri was used, and the enzyme activity were 5.857,5.518,5.996 IU/ml for strep. parasanguinis , Sphingo. paucimobilis , Rhizo. Radiobacter respectively at PH 7, while local isolate E. coli gave best cellulase activity at PH 6.5 reached to 4.963 IU/ml. Efficiency of five isolates were varied in their ability of cellulase activity in liquid media under different temperature, best activity of enzyme for local isolates Sphingo. Paucimobilis and Staph. Sciuri 30 C°, while best enzyme activity was recorded at 25C° for local isolates Strep. Parasanguinis and E. coli, and optimum temperature for enzyme production for Rhizo. Radiobacter was 50 C°. Also aeration and agitation effect was studied for their effect on cellulase activity, and the results showed increasing in activity of enzyme at 50 rpm in local isolates Strep. parasanguinis , Sphingo. paucimobilis , Rhizo. Radiobacter and E. coli while the best enzyme activity for Staph. Sciuri was at 100 rpm. Results also shown that the best productivity of enzyme was after two days of incubation for all isolates.

Identification of acid- and thermotolerant extracellular β-glucosidase activities inZygomycetes fungi

Extracellular beta-glucosidase activity of 94 strains, representing 24 species of the genera Gilbertella, Mucor, Rhizomucor , and Rhizopus was evaluated in submerged culture and under solid state fermentation on wheat bran. Gilbertella persicaria G1 isolate showed the highest activity (70.9 U ml -1 ) followed by other Gilbertella (58.6-59.0 U ml -1 ) and Rhizomucor miehei isolates (29.2-42.0 U ml -1 ). Optimum temperature for enzyme production was 25 degrees C for Gilbertella and Mucor , and 30 degrees C for Rhizomucor and Rhizopus strains. Enzymes of R. miehei strains proved to be thermotolerant preserving up to 92.8% residual activity after heating to 75 degrees C in the presence of cellobiose substrate. Enzymes of Mucor racemosus f. chibinensis, R. miehei and Rhizopus microsporus var. oligosporus strains were activated at acidic condition (pH 4). Glucose was a strong inhibitor for each fungal beta-glucosidase tested but some of them showed ethanol tolerance up to 20% (v/v). Ethanol also activated the enzyme in these strains suggesting glycosyl transferase activity.

Optimization of medium and cultivation conditions forl-amino acid oxidase production byAspergillus fumigatus

A fungal strain was selected from the microbial repository of the North-East Institute of Science and Technology, Jorhat, India, which could produce a high yield of L-amino acid oxidase. 18SrRNA, ITS1, 5.8SrRNA ITS2, and partial 28 S rRNA sequencing and phenotypic characteristics indicate that it belong to the species Aspergillus fumigatus (designated as P13). Maximum production of enzyme (59.55 x 10-3 U/mg dry cell mass) was obtained in a medium containing 10 g/L glucose, 4 g/L yeast extract, and 4 g/L ammonium sulfate, with 20 mmol/L of L-threonine as the inducer. The optimum temperature for enzyme production was 30 degrees C at pH 7.0, with a shaking speed of 200 r/min. At 96 h, the enzyme activity was maximum. The A. fumigatus P13 L-amino acid oxidase accepts a broad substrate range, and the maximum enzyme activity (20.41 x 10-3 U/mg dry cell mass) was obtained with 50 mmol/L of L-tyrosine. In the literature, no reports have been found regarding the production of L-amino acid oxidase by A. fumigatus. The enzyme showed enantiomerically pure amino acid formation, which has tremendous demand in industrial applications.

Isolation and characterization of novel α-amylase from marine Streptomyces sp. D1

In this study, we have reported novel α-amylase enzyme from less extensively studied marine Streptomyces sp. D1. Enzyme production was determined by using media containing 2% sucrose, 0.35% peptone and 0.15% of malt extract. Optimum temperature for enzyme production and activity was found to be 45 °C and enzyme retained almost 50% of its activity at 85 °C. Enzyme activity was also retained in presence of commercially available detergent and oxidizing agents. The partially purified enzyme from strain D1 exhibited specific activity of 113.64 U/mg protein that corresponds to 2.8-fold purification. SDS-PAGE and zymogram activity staining showed a single band equal to molecular mass of 66 kDa. The reported enzyme may have wide spread application for detergent and pharmaceutical industry.

Production of d -hydantoinase by halophilic Pseudomonas sp. NCIM 5109

About 1000 bacterial colonies isolated from sea water were screened for their ability to convert dl-5-phenylhydantoin to d(−)N-carbamoylphenylglycine as a criterion for the determination of hydantoinase activity. The strain M-1, out of 11 hydantoinase-producing strains, exhibited the maximum ability to convert dl-5-phenylhydantoin to d(−)N-carbamoylphenylglycine. The strain M-1 appeared to be a halophilic Pseudomonas sp. according to morphological and physiological characteristics. Optimization of the growth parameters revealed that nutrient broth with 2% NaCl was the preferred medium for both biomass and enzyme production. d-Hydantoinase of strain M-1 was not found to be inducible by the addition of uracil, dihydrouracil, β-alanine etc. The optimum temperature for enzyme production was about 25 °C and the organism showed a broad pH optimum (pH 6.5–9.0) for both biomass and hydantoinase production. The organism seems to have a strict requirement of NaCl for both growth and enzyme production. The optimum pH and temperature of enzyme activity were 9–9.5 and 30 °C respectively. The biotransformation under the alkaline conditions allowed the conversion of 80 g l−1 dl-5-phenylhydantoin to 82 g l−1 d(−)N-carbamoylphenylglycine within 24 h with a molar yield of 93%.

Fermentation kinetics and continuous process of L-asparaginase production.

For the purpose of obtaining L-asparaginase in quantities from Erwinia aroideae, cell growth and enzyme formation were investigated in both batch and continuous fermentation. Using yeast extract as a growth-limiting substrate, the relationship between specific growth rate and substrate concentration was found to fit the Monod equation. The optimum temperature for enzyme production was 24 C, although cell growth was higher at 28 C. The enzyme yield reached its maximum of 4 IU/ml during the negative acceleration growth phase which occurs just prior to stationary growth. Compared to batch fermentations, the continuous fermentation process gave a lower enzyme yield except when the fermentation was conducted at a dilution rate of 0.1 hr(-1). The graphical method frequently used for prediction of continuous fermentation does not apply to L-asparaginase production by E. aroideae. The optimum temperature for enzyme production in continuous process was 24 C, which was the same as in batch process. Increasing the temperature from 24 to 28 C resulted in a 20% loss of enzyme yield.

Fermentation Kinetics and Continuous Process of L-Asparaginase Production

For the purpose of obtaining L-asparaginase in quantities from Erwinia aroideae, cell growth and enzyme formation were investigated in both batch and continuous fermentation. Using yeast extract as a growth-limiting substrate, the relationship between specific growth rate and substrate concentration was found to fit the Monod equation. The optimum temperature for enzyme production was 24 C, although cell growth was higher at 28 C. The enzyme yield reached its maximum of 4 IU/ml during the negative acceleration growth phase which occurs just prior to stationary growth. Compared to batch fermentations, the continuous fermentation process gave a lower enzyme yield except when the fermentation was conducted at a dilution rate of 0.1 hr(-1). The graphical method frequently used for prediction of continuous fermentation does not apply to L-asparaginase production by E. aroideae. The optimum temperature for enzyme production in continuous process was 24 C, which was the same as in batch process. Increasing the temperature from 24 to 28 C resulted in a 20% loss of enzyme yield.