Conditions For Enzyme Production Research Articles

Optimization of L-Glutaminase Enzyme Production by Staphylococcus Aureus Clinical Samples

L-glutaminase produced from staphylococcus aureus. Where, a 108 sample of bacterial isolates collected from clinical source. S. aureus identified by biochemical test and differential medium. The ability of these isolates for L-glutaminase production was examined. Results showed that of S. aureus isolate were L-glutaminase producers with different degrees. Among them S. aureus S22 was the most efficient in enzyme production and the specific activity of L-glutaminase in its crude was 4.1 U/mg. Upon studying the optimum condition for enzyme production.

Isolation, screening and identification of haloarchaea with chitinolytic activity from hypersaline lakes of Iran

Halophiles produce stable enzymes under extreme conditions. The scant information about chitinolytic haloarchaea led us to conduct the present study in order to isolate and screen native halophilic archaea with chitinolytic activity and to optimize the enzyme production conditions. Among 500 haloarchaeal strains isolated from water samples from different hypersaline lakes of Iran, five strains showed chitinolytic activity. Based on biochemical, morphological and molecular analyses, we established that all five potent strains belonged to the genus Natrinema. Besides, observing chitinase function in culture media, through an additional molecular test the presence of the chitinase gene in chitinase-producing strains was also confirmed by PCR amplification. Compared with other potent strains, Natrinema sp. strain BS5 showed significant chitinase production. The production of chitinase in strain BS5 accompanied growth, started at the logarithmic phase and increased to its maximum level at the beginning of the stationary phase. Maximum chitinase production was obtained at 37?C, pH 7.5, 3 M NaCl and 1% colloidal chitin. The strain BS5 showed 38%, 30%, 24% and 28% decreases in enzyme production at 40?C, pH 8, 3.5 M NaCl and 0.5% substrate, respectively. This strain was able to produce the enzyme in NaCl 4 M and in the absence of MgCl2 and MgSO4. This study revealed the strong potential of the genus Natrinema to produce chitinase at high salt concentrations without Mg2+ requirement.

Characterization and optimization of extracellular L-Asparaginase production by selected Actinomycete strain isolated from an algerian wheat bran

L-Asparaginase is an enzyme that hydrolyses the amino acid L-Asparagine into aspartic acid and ammonia. As a medication, L-Asparaginase is used in chemotherapy to treat acute lymphoblastic leukaemia by depleting circulating Asparagine and depriving tumor cells. Interest in Actinomycetes as potential producers of antibiotics and enzymes encouraged us to investigate an isolated strain (CA01) from soft wheat bran.The Actinomycete strain was characterized based on its morphological and biochemical characteristics and selected due to a proved promising ability to produce L-Asparaginase optimized in both solid and liquid media cultures.The conditions of enzyme production were standardized according to a one-factor-at-a-time (OFAT) experimental design.To obtain optimal medium combination, a Box-Behnken Response Surface Methodology (RSM) has been adopted by choosing the most influential factors. The optimal conditions for the enzyme production were (g/l): L-Asparagine 10.7; Glucose 2.7; starch 7, in based medium containing (g/l): K2HPO4 0.5; MgSO4, 7H2O 0.1, corresponding to an optimal enzymatic activity of 8.03 IU/ml at 27.83°C. The maximum production of enzyme was reached on the sixth day of experiment. The ANOVA test (P value ˂ 0.05) and adjusted R2 values close to the experimental R2 show that the obtained model of the active L-Asparaginase of CA01 strain production is significant with the following linear terms: temperature, substrate concentration, Glucose concentration and there squared.

Optimization of the Enzyme Production Conditions of Bacillus licheniformis and Its Effect on the Degradation of Corn Straw

Optimization of the Enzyme Production Conditions of <i>Bacillus licheniformis</i> and Its Effect on the Degradation of Corn Straw

Comparison of ionic liquids for efficient pretreatment of brewery spent grain

A newly isolated fungus Aspergillus niger RCKH-3 was employed for cost effective production of a complete cellulase system under solid state fermentation (SSF). Overall evaluation criteria approach (OEC) was used as a novel approach for medium engineering to maximize the cellulase production. Among several factors studied, NH4Cl, inoculum age and temperature contributed maximally towards cellulase production. The highest interactions were observed between inoculum age and beef extract, followed by substrate to moisture ratio and PEG 8000. The statistical model provided optimal enzyme production conditions as: 5 g wheat bran; mineral salt solution (pH 5) and containing (g/L); NH4Cl (0.5), beef extract (2.0) and PEG 8000 (3.0) substrate to moisture ratio of 1:3.5 and 30% (v/w) 18 h old inoculum. Under optimum conditions, cellulase production experienced 2.5, 3.0 and 1.5 fold enhancements in FPase, CMCase and β-glucosidase activities, respectively. The estimated production costs ($ per 1000 units) for FPase, CMCase, β-glucosidase were 0.47, 0.21 and 0.04, respectively. Saccharification of office waste paper using crude cellulase resulted in reducing sugar yield of 534 mg/g dry substrate. Cellulase of newly isolated fungi A. niger RCKH-3 shows potential in biofuel applications based on its comparatively higher saccharification abilities and lower production costs.

STUDY OF ALPHA-AMYLASE OBTAINED BY SOLID STATE FERMENTATION OF CASSAVA RESIDUE IN AQUEOUS TWO-PHASE SYSTEMS

Abstract The α-amylase enzyme was produced by solid state fermentation (SSF) and its partition behavior in Aqueous Two-Phase Systems (ATPS) was determined using systems consisting of polyethylene glycol (PEG) and potassium phosphate buffer. To determine the best conditions for α-amylase production, a central composite design was constructed for two independent variables: fermentation time and moisture content, and for the partition study, a face-centered central composite design (CCF) with four independent variables: PEG molecular weight, pH, temperature and partition time. It was indicated that the best enzyme production conditions were a fermentation time of 35.0 hours and 45.7% moisture content. From the results obtained, it was found that increasing the polymer molecular weight promotes a reduction of the enzyme concentration in the upper phase, increasing its concentration in the lower phase. It was also observed that the partition coefficients increased with increasing pH values and a reduction in temperature.

Biodegradability of polyethylene by bacteria and fungi from Dandora dumpsite Nairobi-Kenya.

This study aimed at isolating and identifying bacteria and fungi with the capacity to degrade low density polyethylene (LDPE). The level of biodegradation of LDPE sheets with bacterial and fungal inoculums from different sampling points of Dandora dumpsite was evaluated under laboratory conditions. Incubation of the LDPE sheets was done for sixteen weeks at 37°C and 28°C for bacteria and fungi respectively in a shaker incubator. Isolation of effective candidates for biodegradation was done based on the recorded biodegradation outcomes. The extent of biodegradation on the polyethylene sheets was assessed by various techniques including weight loss analysis, Fourier Transform Infrared Spectroscopy (FTIR) and GC-MS. Fourier Transform Infra-Red spectroscopy (FTIR) analysis revealed the appearance of new functional groups attributed to hydrocarbon degradation after incubation with the bacteria and fungi. Analysis of the 16S rDNA and 18S rDNA sequences for bacteria and fungi respectively showed that bacteria belonging to genera Pseudomonas, Bacillus, Brevibacillus, Cellulosimicrobium, Lysinibacillus and fungi of genus Aspergillus were implicated as polyethylene degraders. An overall analysis confirmed that fungi are generally better degraders of polyethylene than bacteria. The highest fungal degradation activity was a mean weight reduction of 36.4±5.53% attributed to Aspergillus oryzae strain A5, 1 (MG779508). The highest degradation activity for bacteria was a mean of 35.72± 4.01% and 20.28± 2.30% attributed to Bacillus cereus strain A5,a (MG645264) and Brevibacillus borstelensis strain B2,2 (MG645267) respectively. Genus Aspergillus, Bacillus and Brevibacillus were confirmed to be good candidates for Low Density Poly Ethene bio-degradation. This was further confirmed by the appearance of the aldehyde, ether and carboxyl functional groups after FTIR analysis of the polythene sheets and the appearance of a ketone which is also an intermediary product in the culture media. To improve this degrading capacity through assessment of optimum conditions for microbial activity and enzyme production will enable these findings to be applied commercially and on a larger scale.

Cost effective production of complete cellulase system by newly isolated Aspergillus niger RCKH-3 for efficient enzymatic saccharification: Medium engineering by overall evaluation criteria approach (OEC)

A newly isolated fungus Aspergillus niger RCKH-3 was employed for cost effective production of a complete cellulase system under solid state fermentation (SSF). Overall evaluation criteria approach (OEC) was used as a novel approach for medium engineering to maximize the cellulase production. Among several factors studied, NH4Cl, inoculum age and temperature contributed maximally towards cellulase production. The highest interactions were observed between inoculum age and beef extract, followed by substrate to moisture ratio and PEG 8000. The statistical model provided optimal enzyme production conditions as: 5 g wheat bran; mineral salt solution (pH 5) and containing (g/L); NH4Cl (0.5), beef extract (2.0) and PEG 8000 (3.0) substrate to moisture ratio of 1:3.5 and 30% (v/w) 18 h old inoculum. Under optimum conditions, cellulase production experienced 2.5, 3.0 and 1.5 fold enhancements in FPase, CMCase and β-glucosidase activities, respectively. The estimated production costs ($ per 1000 units) for FPase, CMCase, β-glucosidase were 0.47, 0.21 and 0.04, respectively. Saccharification of office waste paper using crude cellulase resulted in reducing sugar yield of 534 mg/g dry substrate. Cellulase of newly isolated fungi A. niger RCKH-3 shows potential in biofuel applications based on its comparatively higher saccharification abilities and lower production costs.

Exploring PTDH-P450BM3 Variants for the Synthesis of Drug Metabolites.

The conversion of a series of pharmaceutical compounds was examined with three variants of cytochrome P450BM3 fused to phosphite dehydrogenase (PTDH) to enable cofactor recycling. Conditions for enzyme production were optimized, and the purified PTDH-P450BM3 variants were tested against 32 commercial drugs by using rapid UPLC-MS analysis. The sets of mutations (R47L/F87V/L188Q and R47L/F87V/L188Q/E267V/G415S) improved conversion for all compounds, and a variety of products were detected. Product analysis showed that reaction types included C-hydroxylation, N-oxidation, demethylation, and aromatization. Interestingly, enzymatic aromatization could occur independent of the addition of reducing coenzyme. These results identified new conversions catalyzed by P450BM3 variants and showed that a small set of mutations in the oxygenase domain could broaden both substrate range and reaction type.

Optimization of Arginine deiminase production from a local higher productive isolate Enterococcus faecium M1

Arginine deiminase (ADI) is an important enzyme in many biological applications and a cancer treatment agent, recently increasing attention is focused to choose a higher productive strain from different bacterial sources. Eighty nine Enterococcus isolates were obtained from 215 samples (20 clinical UTI specimens, 52 human stool specimens, 6 soil and 11 from sewage water), fifty seven isolates were ADI producer. Twenty five isolates were more active in enzyme production, fourteen of them were identified as E. faecium and 11 isolates were E. faecalis. E. faecium isolate M1 obtained from UTI was the most efficient in ADI production, the specific activity of ADI produced from this isolate was 2.64U/mg protein. The effect of culture medium (Mineral salt broth) components on ADI production and other cultural conditions were determined to find the optimum conditions for enzyme production. The maximum ADI production was achieved when the medium was supplemented with 20mM arginine, 1% sucrose, 1% casein, pH 7.5 and incubated at 37ºC for 18 hours. Under these conditions, the specific activity of ADI was 5.1U/mg protein. By concluding, the present study was designed to select ADI higher productive isolate and optimum culture conditions for increasing the production of this enzyme in order to use it as a potent cancer treatment agent after purifying and characterizing it in the future.

Single cell oil production by Trichosporon cutaneum and lignocellulosic residues bioconversion for biodiesel synthesis

Abstract Barley hull hydrolysate was used as the fermentation feedstock of Trichosporon cutaneum CTM-30125 yeast strain for production of microbial lipid as the potential raw material of biodiesel. An effective hydrolysis process based on the combination of dilute acid hydrolysis and enzymatic saccharification was developed to maximize recovery of total sugars. On synthetic media, CTM-30125 can assimilate glucose and xylose simultaneously and accumulated intracellular lipid up to 43.6% (w/w). However, when grown on acid barley hull hydrolysate (ABHH) the lipid content was low (16.6% w/w) and a detoxification step was required. Overliming by Ca(OH)2 treatment and adsorption by activated charcoal improved the fermentability of ABHH significantly by removing the inhibitors and reducing the nitrogen content. The solid fraction resulting from the acid treatment of the barley hull was treated with hydrolytic enzyme complex produced by the hyper-cellulolytic fungal strain Trichoderma reesei Rut C-30. Enzyme production and hydrolytic conditions were optimized to enhance the saccharification. The structural changes in native and treated raw material were monitored using FTIR spectra. Detoxified ABHH and enzymatic hydrolysate were successively fermented by the CTM-30125 strain in fed-batch bioreactor without addition of external nitrogen source. A total biomass of 17.5 g/L with a lipid content of 38.2% (corresponding to a lipid yield of 6.7 g/L) could be achieved after 168 h of culture. The relative fatty acid composition indicates the presence of increased levels of saturated (41.2%) and monounsaturated (52.5%) fatty acids in the oil, making it ideal for biodiesel production.

Application of Box-Behnken Design for the Optimization of Culture Conditions for Novel Fibrinolytic Enzyme Production by Bacillus altitudinis S-CSR 0020

Application of Box-Behnken Design for the Optimization of Culture Conditions for Novel Fibrinolytic Enzyme Production by Bacillus altitudinis S-CSR 0020

&lt;b&gt;Production of cellulases by &lt;i&gt;Penicillium oxalicum&lt;/i&gt; through solid state fermentation using agroindustrial substrates

The purpose of this study was to define the factors that influence the production of cellulases by Penicillium oxalicum , a strain obtained from a leaf-cutting ant colony and identified based on the ITS gene . The experimental design included solid state fermentation using sugarcane bagasse and lignocellulosic sorghum as the lignocellulosic substrate. The variables were analyzed using a 2 5-1 fractional factorial design, with three replicates on the central point. All the variables analyzed influenced the production of at least one of the three cellulose types analyzed. The highest values observed were: FPase 4.2 U g -1 , CMCase 9.2 U g -1 and Avicelase 8.4 U g -1 using lignocellulosic sorghum as the substrate. The best conditions for enzyme production were: incubation temperature at 40oC, initial moisture of 60%, pH of 4.0 and a growth period of four days using lignocellulosic sorghum as the substrate.

Production and Optimization of Amylase and Glucoamylase from Aspergillus Niger under Solid State Fermentation for Effective Production of Glucose Syrup

The continual demand for sugars and coupled with the chemical hazards associated with chemically produced syrup had led to the development of enzymatic technology which has increased the production of glucose syrups most especially using enzymes from microbial origin. The aim of this work was to isolate, screen and optimized the best cultural condition that can lead to the production of glucose syrups from Nigeria cassava starch using enzymes produced by soil microorganisms. Also to determine proximate and physiochemical properties of the glucose syrup produced from yellow and white maize using microbial enzymes. The results obtained from the preliminary screening showed that most of the isolates obtained were Aspergillus niger. It was observed that Aspergillus niger had the largest zone of amylase activity of 35.0 mm when compared to Fusarium pallidorosium that had the lowest (5.0 mm). The optimal conditions for enzyme production; temperature, incubation period, inoculum concentration, and pH were 30 oC, 5 days, 1 × 106 CFU/ml, and pH of 5 respectively for the production of glucoamylase and amylase. Generally, strain AMO1 was observed to produced more glucoamylase when compare to amylase production. The crude protein, the colour, viscosity, ash content, reducing sugar content, pH and carbohydrate contents of the syrup from yellow maize were more than that of the syrup from white maize.&#x0D;

Effect of Physical and Chemical Parameters on the Activity of Purified Phosphatase Enzyme Produced by Bacillus cereus

Phosphorus is one of the most important nutrients for plant growth and development. This study aimed to survey the optimum condition for phosphatase enzyme production, purified and studied its properties. Phosphatase enzyme activity was determined by end point method. Phosphatase enzyme was obtained from Bacillus cereus EME 66 isolate. The enzyme was purified and characterized, using a three-step purification procedure with 12.3-fold. The phosphatase enzyme was partially purified using ammonium sulfate fractionation followed by ultrafiltration. The acid phosphatase is a monomer protein purified gel filtration to 5.6 fold. Results showed that the optimum temperature for the purified enzyme activity was 60 °C and it was stable at temperatures below 60 °C. This enzyme was stable between pH 4.0-6.0, and the optimal pH activity was found to 5.0. The activity of the enzyme enhanced by heavy metals (Fe 3+ , Cu 2+ , Ca 2+ , Mg 2+ and K + ). The enzyme activity was strongly inhibited by heavy metals Zn 2+ . The present article reveals on enzymatic characterization of acid phosphatase enzyme.Â

A novel extracellular low-temperature active phytase from Bacillus aryabhattai RS1 with potential application in plant growth.

Bacillus aryabhattai RS1 isolated from rhizosphere produced an extracellular, low temperature active phytase. The cultural conditions for enzyme production were optimized to obtain 35 UmL-1 of activity. Purified phytase had specific activity and molecular weight of 72.97 Umg-1 and ∼40 kDa, respectively. The enzyme was optimally active at pH 6.5 and 40°C and was highly specific to phytate. It exhibited higher catalytic activity at low temperature, retaining over 40% activity at 10°C. Phytase was more thermostable in presence of Ca2+ ion and retained 100% residual activity on preincubation at 20-50°C for 30 min. Partial phytase encoding gene, phyB (816 bp) was cloned and sequenced. The encoded amino acid sequence (272 aa) contained two conserved motifs, DA[A/T/E]DDPA[I/L/V]W and NN[V/I]D[I/L/V]R[Y/D/Q] of β-propellar phytase and had lower sequence homology with other Bacillus phytases, indicating its novelty. Phytase and the bacterial inoculum were effective in improving germination and growth of chickpea seedlings under phosphate limiting condition. Moreover, the potential applications of the enzyme with relatively high activity at lower temperatures (20-30°C) could also be extended to aquaculture and food processing. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:633-641, 2017.

OPTIMIZATION OF BIOMASS AND ENDO-B-1,4-GLUCANASE BY WHITE ROT FUNGI NATIVE FROM ARGENTINA

A large number of microorganisms are capable of producing cellulases; however, fungi are considered among the most active enzymes producers. Different strains of white rot fungi (WRF) were reported to have a great potential for biotechnological applications and cellulase enzymes is much involved in this effect, especially for bioethanol production. Misiones is included in a forest region characterized by a vast biodiversity, and within this environment, a large number of microorganisms can be isolated, including numerous white rot fungi. In the present work, response surface methodology (RSM) was applied for the determination of biomass and endo-?-1,4-glucanase activity factors dependence from three Argentinean WRF in submerged cultures. RSM was carried out with a Box-Behnken design using 3 variables, temperature (25, 29 and 33 C), incubation time (7, 10.5 and 14 days) and pH (3.5, 4.5 and 5.5) with three central points and 2 replications. The best conditions for enzymatic activity were achieved at maximum level of temperature (33 C), low level of pH (3.5) and intermediate level of incubation time (10.5 h). We have presented here, preliminary studies on biomass and enzyme production of three fungi. Our data show that we have not yet reached the optimal enzyme production conditions, so further experiments are needed to reach optimal conditions.

Optimization of the production conditions of the lipase produced by Bacillus cereus from rice flour through Plackett-Burman Design (PBD) and response surface methodology (RSM)

In this study, the screening of lipase positive bacteria from rice flour was carried out by Rhodamin B agar plate method. Bacillus cereus was identified by 16S rDNA method. Screening of the appropriate variables and optimization of the lipase production was performed using Plackett-Burman design (PBD) and response surface methodology (RSM). Among the isolated bacteria, an aerobic Bacillus cereus strain was recognized as the best lipase-producing bacteria (177.3 ± 20 U/ml). Given the results, the optimal enzyme production conditions were achieved with coriander seed extract (CSE)/yeast extract ratio of 16.9 w/w, olive oil (OO) and MgCl2 concentration of 2.37 g/L and 24.23 mM, respectively. In these conditions, the lipase activity (LA) was predicted 343 U/mL that was approximately close to the predicted value (324 U/mL), which was increased 1.83 fold LA compared with the non-optimized lipase. The kinetic parameters of Vmax and Km for the lipase were measured 0.367 μM/min.mL and 5.3 mM, respectively. The lipase producing Bacillus cereus was isolated and RSM was used for the optimization of enzyme production. The CSE/yeast extract ratio of 16.9 w/w, OO concentration of 2.37 g/L and MgCl2 concentration of 24.23 mM, were found to be the optimal conditions of the enzyme production process. LA at optimal enzyme production conditions was observed 1.83 times more than the non-optimal conditions. Ultimately, it can be concluded that the isolated B. cereus from rice flour is a proper source of lipase.

Optimization of liquid-state fermentation conditions for the glyphosate degradation enzyme production of strain Aspergillus oryzae by ultraviolet mutagenesis

ABSTRACTThis study aimed to obtain strains with high glyphosate-degrading ability and improve the ability of glyphosate degradation enzyme by the optimization of fermentation conditions. Spore from Aspergillus oryzae A-F02 was subjected to ultraviolet mutagenesis. Single-factor experiment and response surface methodology were used to optimize glyphosate degradation enzyme production from mutant strain by liquid-state fermentation. Four mutant strains were obtained and named as FUJX 001, FUJX 002, FUJX 003, and FUJX 004, in which FUJX 001 gave the highest total enzyme activity. Starch concentration at 0.56%, GP concentration at 1,370 mg/l, initial pH at 6.8, and temperature at 30°C were the optimum conditions for the improved glyphosate degradation endoenzyme production of A. oryzae FUJX 001. Under these conditions, the experimental endoenzyme activity was 784.15 U/100 ml fermentation liquor. The result (784.15 U/100 ml fermentation liquor) was approximately 14-fold higher than that of the original strain. The result highlights the potential of glyphosate degradation enzyme to degrade glyphosate.

Optimization of production, biochemical characterization and in vitro evaluation of the therapeutic potential of fibrinolytic enzymes from a new Bacillus amyloliquefaciens

The capacity of fibrinolytic enzymes to degrade blood clots makes them of high relevance in medicine and in the pharmaceutical industry. In this work, forty-three microorganisms of the genus Bacillus were evaluated for their potential to produce fibrinolytic proteases. Thirty bacteria were confirmed as producers of fibrinolytic enzymes, the best results obtained for the strain Bacillus amyloliquefaciens UFPEDA 485. The optimization of the enzyme production conditions was done by a central composite design (CCD) star 23 that allowed to define the optimal conditions for soybean flour and glucose concentrations and agitation rate. The highest fibrinolytic activity (FA) of 813 U mL–1 and a degradation of blood clot in vitro of 62% were obtained in a medium with 2% (w/v) of soybean flour and 1% (w/v) glucose at 200 rpm after 48 h of cultivation, at pH 7.2 and 37 °C. The obtained fibrinolytic enzyme was characterized biochemically. Fibrinolytic activity was inhibited by PMSF (fluoride methylphenylsulfonyl - C7H7FO2S) 91.52% and EDTA (ethylenediaminetetraacetic acid - C10H16N2O8) 89.4%, confirming to be a serine-metallo protease. The optimum pH and temperature were 7.0 and 37 oC, respectively, and the enzyme was stable for 12 h. The fibrinolytic activity at physiological conditions of this enzyme produced by Bacillus amyloliquefaciens UFPEDA 485, as well as its long term stability, demonstrate that it has suitable characteristics for human and veterinary applications, and promises to be a powerful drug for the treatment of vascular diseases.