Maximum Enzyme Production Research Articles

Cellulase Producing Bacteria Isolation, Screening and Media Optimization from Local Soil Sample

Cellulase enzyme earns consecutively increasing demand in industries of Bangladesh. This experiment has been planned to develop the enzyme industry in our country rather than import enzymes from other countries and conclude with isolating and screening cellulose-degrading bacteria from local land samples and ensuring maximum enzyme production through media optimization. After collecting soil samples from various Tangail areas, a typical dilution of series was performed on a Carboxymethylcellulose plate (CMC) to produce cellulase-producing microorganisms. Based on the hydrolysis zone of the ratio, four isolates were chosen and labeled CBM21, CSG8, CSR35, and CHT8 for further morphological and biochemical analysis. Then subjected to cellulase production in 250 ml of Erlenmeyer flask using CMC broth for 48 hours of fermentation at 37°C at an agitation rate of 140rpm, and cellulase enzyme assay methods evaluated crude enzymes. Eventually, those confirmed isolates were optimized using different parameters by submerged fermentation process for enhanced cellulase production. Among four isolates, CSG8 and CBM21 exhibited the maximum clearance zone with the hydrolytic values 6.96 and 8.46. These isolates were identified as Pseudomonas spp. (CSG8). According to the screening process, CBM21 (0.156U/ml), CSG08 (0.106U/ml), and E. coli (negative control) have been used for optimization with their initial enzyme productivity. After optimization, an amazing enhancement was noticed for CBM21 (1.35 U/ml) and CSG8 (1.23 U/ml) compared to E.coli (0.44 U/ml). The enzyme was characterized in isolates with an effective temperature of 40 and 35°C for CBM21 and CSG8, respectively, and pH 7, glucose 5% with the incubation time 24h was optimal. Aftermath, this type of optimization can be a significant initiative in the enzyme industries of Bangladesh.

Nano-metals forming bacteria in Egypt. I. Synthesis, characterization and effect on some phytopathogenic bacteria in vitro

Bacterial metal reducers were isolated from water samples collected from harsh condition locations in Egypt. Four selected isolates were identified as Enterococcus thailandicus, Pseudomonas putida, Marinobacter hydrocarbonoclasticus, and P. geniculata for Copper (Cu), Iron (Fe), Cobalt (Co) and Zinc (Zn) Nanoparticles (NPs) production sequentially. Nitrate reductase enzyme was assayed for bacterial isolates which demonstrated that P. putida, and M. hydrocarbonoclasticus have the maximum enzyme production. The produced NPs were characterized by using XRD, TEM, UV–VIS spectroscopy. Magnetic properties for all selected metals NPs were measured using Vibrating Sample Magnetometer (VSM) and demonstrated that FeNPs recorded the highest magnetization value. The antibacterial activity of selected metals NPs was tested against some phytopathogenic bacteria causing the following diseases: soft rot (Pectobacterium carotovorum, Enterobacter cloacae), blackleg (Pectobacterium atrosepticum and Dickeya solani), brown rot (Ralstonia solanacearum), fire blight (Erwinia amylovora) and crown gall (Agrobacterium tumefaciens). All metals NPs showed an antagonistic effect against the tested isolates, particularly, FeNPs showed the highest antibacterial activity followed by CuNPs, and ZnNPs. Due to the small size, high reactivity, and large surface area of biologically synthesized NPs, they are used as a good disinfector, and can be considered as a new and alternative approach to traditional disease management methods.

A New Extracellular β-Galactosidase Producing Kluyveromyces sp. PCH397 from Yak Milk and Its Applications for Lactose Hydrolysis and Prebiotics Synthesis.

The online version contains supplementary material available at 10.1007/s12088-021-00955-1.

Optimization of α‐amylase and Glucoamylase Production Using Cull Potatoes as Substrate

AbstractAmylases are required for the conversion of starchy substrate into sugars. Commercially available enzymes are quite costly which makes the process uneconomical. This study is conducted to standardize physico‐chemical parameters such as substrate concentration (1.0–5.0%), temperature (25–40 °C), pH (4.0–7.0), incubation time (18–24) h for α‐amylase and 3–6 days for glucoamylase production) for optimum production of α‐amylase from bacterial isolate (AM1) and glucoamylase from Aspergillus niger by using cull potatoes as a substrate. Various organic and inorganic nitrogen sources (urea, yeast extract and soybean meal, and ammonium sulphate) are tried for maximum enzyme production. Among the four nitrogen sources, yeast extract supported maximum α‐amylase and glucoamylase production. Among the different amylolytic isolates from fermented jalebi dough and AM1culture, maximum α‐amylase activity is shown by AM1 (90.46 U mL−1) as compared to J1 (71.02 U mL−1). After optimization, α‐amylase activity of AM1 increases by 41.73% from 198.20 to 280.92 U mL−1 at 5.0% substrate concentration, 30 °C temperature, 24 h of incubation and pH 5.0. Glucoamylase activity of A. niger increases by 38.78% from 228.72 to 317.44 U mL−1 after optimization at 5% substrate concentration, pH 5, incubation period of 4 days and temperature 30 °C.

Valorization of corn stover and molasses for enzyme synthesis, lignocellulosic hydrolysis and bioethanol production by Hymenobacter sp. CKS3

During the last few decades, energy demand is increasing rapidly. Concerning this, the use of renewables — lignocellulose biomass, for bioethanol production, as an efficient alternative to replacing fossil fuels, is highly recommended. In this study, valorization of two agricultural wastes was used for various hydrolytic enzyme production by Hymenobacter sp. CKS3, lignocellulosic hydrolysis, and bioethanol production. Conditions for obtaining maximum enzyme production, using agro-industrial waste — molasses and corn stover, were statistically optimized. Under the optimal conditions, in a medium containing 5.0% corn stover, 2.5% molasses, and during 94.55 h (∼4 days) of fermentation, the maximum enzymatic activity was achieved - CMCase 1.11 IU/ml, Avicelase 0.92 IU/ml, and pectinase 3.69 IU/ml. The obtained crude enzyme mixture was further used for enzymatic hydrolysis of non-treated corn stover and bioethanol production. The reducing sugar yield of 3.85 g/l was obtained under optimal conditions (corn stover 6.6% and time of hydrolysis 78.8 h (∼3 days and 7 h)). Scanning electron microscopy revealed structural changes in corn stover samples after enzymatic hydrolysis. Under non-optimized conditions, 0.37% of ethanol was produced by waste brewer’s yeast. The obtained results show that bacteria belonging to the genus Hymenobacter have a still unexplored enzymatic potential that could be used for sustainable production of biotechnologically value-added products including biofuels. Specifically, for the first time, a soil bacterium, classified within the genus Hymenobacter, was used for cellulases (CMCase and Avicelase) and pectinase production.

Optimization of Cultural Conditions for Maximum Production of Fibrinolytic Enzymes from the Local Marine Bacterial Isolates and Evaluation of their Wound Healing and Clot Dissolving Properties

Fibrinolytic enzymes find necessary applications to treat and prevent cardiovascular diseases. In this study, optimal conditions for enhancing the production of fibrinolytic enzyme from local marine bacterial strains were evaluated. The present study also focuses on screening of wound healing efficacy of the isolated fibrinolytic enzymes.Various physical parameters such as temperature, pH, incubation time and medium components viz. inoculum size, substrate (nitrogen and carbon) concentrations were optimized. A cultivation medium was designed using optimized conditions for mass production of fibrinolytic enzyme and specific activity of enzyme was analyzed. The maximum enzyme production was observed at 37 °C temperature, 8.0 pH,substrate concentration with 3 ml inoculum size and 32 h. of incubation time. Among the different carbon sources tested, Mannitol showed maximum enzyme activity i.e 538 U/ml. yeast extract was found to be the best nitrogen source with an enzyme activity of 498 U/ml. The best substrate for the production fibrinolytic enzyme was found to be kernelwith high activity of 1056U/ml. The crude enzyme displayed potent activity and digested blood clot completely in in vitro condition and exhibited potent activity on wound healing property in macrophages.

Optimization of culture conditions for production of phytase by Aspergillus niger

In this research, the environmental and nutritional factors that control the improvement of phytase enzyme production from the two highest phytase producing fungi Aspergillus niger isolate no 62 and Aspergillus terreus isolate no 24 out of 99 isolated fungi from local soil samples were studied. It was found to have the highest enzyme activity after 8 days of incubation period, temperature of 30 ° C, initial pH of 6 in case of Aspergillus niger isolate no 62 and 12 days, temperature of 25 ° C and initial pH of 6.5 in case of Aspergillus terreus isolate no 24, where It was found that these are the best environmental conditions for maximum enzyme production. The Aspergillus niger isolate no 62 was chosen as the best phytase producing fungi. By studying the effect of different nutritional factors, it was found that sucrose was the best source of carbon, giving the highest enzyme activity at an optimal concentration of 1.5% (weight / volume), and ammonium nitrate when used at a concentration of 0.6% (weight / volume) was found to be the best source of nitrogen to produce phytase enzyme. When studying the effect of different concentrations of the substrate on enzyme production, it was found that when used at a concentration of 3 g / L, the substrate showed maximum phytase production. The descending sequence of agricultural waste according to phytase production by A. niger is as follows: wheat bran> rice bran> cotton stems> broad beans> cotton stems> pomegranate pulp> pea husks.

The glucoamylase from Aspergillus wentii: Purification and characterization.

This study describes for the first time the purification and characterization of a glucoamylase from Aspergillus wentii (strain PG18), a species of the Aspergillus genus Cremei section. Maximum enzyme production (∼3.5 U/ml) was obtained in submerged culture (72 h) with starch as the carbon source, at 25°C, and with orbital agitation (100 rpm). The enzyme was purified with one-step molecular exclusion chromatography. The 86 kDa purified enzyme hydrolyzed starch in a zymogram and had activity against p-nitrophenyl α- d-glucopyranoside. The optimal enzyme pH and temperature were 5.0 and 60°C (at pH 5.0), respectively. The Tm of the purified enzyme was 60°C, at pH 7.0. The purified glucoamylase had a KM for starch of 1.4 mg/ml and a Vmax of 0.057 mg/min of hydrolyzed starch. Molybdenum activated the purified enzyme, and sodium dodecyl sulfate inhibited it. A thin layer chromatography analysis revealed glucose as the enzyme's main starch hydrolysis product. An enzyme's peptide sequence was obtained by mass spectrometry and used to retrieve a glucoamylase within the annotated genome of A. wentii v1.0. An in silico structural model revealed a N-terminal glycosyl hydrolases family 15 (GH15) domain, which is ligated by a linker to a C-terminal carbohydrate-binding module (CBM) from the CBM20 family.

Carbon and Nitrogen Sources Effect on Pectinase Synthesis by Aspergillus niger Under Submerged Fermentation

Pectinases are the commercial enzymes that are abundantly employed in various industries like fruit juice industries for clarification, wine indutsry and paper industry for bleaching up pulp. The present work was done on culture conditions optimization for production of pectinases under submerged fermentation using wheat bran as a substrate. Fungal strains were isolated from vegetable waste dump yard soils of Warangal district of Telangana state and screened for their activity on pectin agar medium. Among 30 isolates, two fungal strains showed good activity and identified them as A. niger and A. flavus. The effects of the different carbon and nitrogen sources on pectinases viz. exo-PG, endo-PG, endo-PL and PME by A. niger with 1% wheat bran was carried out in submerged fermentation. These studies revealed that carbon and nitrogen sources have shown considerable influence on enzyme production. Among all the carbon sources tried, sucrose at 1% was shown to be efficient carbon source for all four types of pectinases under investigation. For endo-PG, endo-PL and PME maximum enzyme production were recorded on 8th day of incubation period but for exo-PG enhanced production was observed on 12th day. A. niger could not produce PME on 12th day from 2.50% to subsequent concentrations. Among nine different nitrogen sources were screened, maximum pectinase production was recorded in sodium nitrate at 0.2 % for A. niger. Endo-PG, endo-PL and PME maximum production were recorded on 8th day of incubation and for exo-PG maximum production was observed on 12th day. No PME production was observed in A. niger on 12th day.

Effect of alkaline protease produced from fish waste as substrate by Bacillus clausii on destaining of blood stained fabric

Alkaline protease or peptidases are the largest groups of enzymes in the biological industry with a variety of application in manufacturing units used in the process of substrate stabilization, dehairing, diagnosis, extraction, food production, destaining, etc., where the pH of the environmental conditions remain above neutral pH. Because of these wider applications of alkaline proteases in industries, their demand is increasing to compete with their chemical counterpart. An alkaline tolerant bacterial strain Bacillus clausii wasisolated from fish waste and used for mass production of alkaline protease using fish waste homogenate as media. Preliminary study on optimization of conditions for the mass production carried out. The optimum temperature for production ranges between 25°C and 35°C and pH determined as 9. The mass production of extracellular alkaline protease carried out using mobilized and immobilized cells of B. clausii at optimized condition using production media, the mixture of production media and fish waste homogenate and in nutrient broth. The recorded results showed that the maximum enzyme production obtained with immobilized cells in nutrient broth media and followed by fish waste homogenate media of 8900 U/ml and 8600 U/ml. Purification protease enzyme yielded 0.35 g/ml from the production media . Bloodstained cloth treated with immobilized enzyme removed the stain completely compared to treatment with non-immobilized enzyme and commercially used detergent. So, the current study suggests the usage of microbial alkaline protease in household detergents to replace the usage of synthetic detergents and save the environment from chemical pollutants.

α-Amylase and cellulase production by novel halotolerant Bacillus sp.PM06 isolated from sugarcane pressmud.

A novel Bacillus sp.PM06 isolated from sugarcane waste pressmud was tested for extracellular α-amylase and cellulase enzyme production. The effect of different substrates, nitrogen sources, pH, and temperature on growth and extracellular enzyme production was examined. Bacillus sp.PM06 was able to grow with starch and carboxymethyl cellulose (CMC) as a sole source of carbon and ammonium chloride was found to be the best nitrogen source. Maximum enzyme production was obtained at 48H for both α-amylase and cellulase. The optimal condition for measuring enzyme activity was found to be pH 5.5 at 50°C for α-amylase and pH 6.4 at 60°C for cellulase respectively. It was found that Bacillus sp.PM06 exhibited halotolerance up to 2M Sodium chloride (NaCl) and Potassium chloride (KCl). The isolate could produce α-amylase in the presence of 2M NaCl and 1M KCl. However, the strain produced cellulase even in the presence of 2M NaCl and KCl. Concomitant production of both enzymes was observed when the medium was supplemented with starch and CMC. A maximum of 31±1.15U/mL of amylase and 15±1.5U/mL of cellulase was produced in 48H. The enzyme was partially purified by Ammonium sulphate (NH4 )2 SO4 precipitation with 2.2 and 2.3-fold purification.

Utilization of orange pulp and corn steep liquor for L-methioninase production by Wickerhamomyces subpelliculosus

Background and objective L-methioninase has attracted much attention with respect to its proposed applications in both pharmaceuticals and food industry. The aim of this study was to develop an economic medium formulation using agro-industrial by-products as substrates for large-scale production of L-methioninase. Materials and methods Identification of a high L-methioninase-producing yeast isolate was carried out using 18S rRNA molecular technique. Screening of various agro-industrial by-products and optimization of different process parameters were investigated. Partial purification and characterization of a crude enzyme were carried out. Results and discussion A high L-methioninase-producing yeast isolate was phylogenetically identified as Wickerhamomyces subpelliculosus. Among different agro-industrial by-products tested, orange pulp supported maximum enzyme production (94.08 U/ml) followed by cane and beet molasses. In addition, corn steep liquor (CSL) gave high enzyme level (141.12 U/ml) and could be used as an inexpensive alternate for yeast extract. The optimum growth conditions were found to be orange pulp 30% (w/v), CSL 4% (v/v), CaCl2 0.05%, and KH2PO4 0.05% (w/v) at pH 6.0 after 48 h of incubation. This developed medium formulation increased L-methioninase production (161.95 U/ml) by twofold compared with that obtained by the Czapek–Dox’s medium (73.92 U/ml). Crude enzyme was partially purified by heat treatment at 70°C with 2.9 purification fold. The enzyme activity was optimal at temperature 60°C and pH 7.0. The results showed that a mixed formulation of orange pulp and CSL can be used as an effective and economic substrate for the production of L-methioninase by W. subpelliculosus.

Isolation, Screening and Time Course Study of Amylase-producing Fungi from Garri Processing Environment

Amylases (E.C.3.2.1.1) are enzymes which catalyze the breakdown of -1,4 glycosidic linkages of starch to simple sugars and different monomeric products. Microbial amylases are essential enzymes compared with animals and plants amylases in industrial applications. Thus, in southern Nigeria, garri processing waste-water from mills poses a serious environmental challenge and this situation could be exploited by utilizing microorganisms colonizing the area to manufacture microbial products. This present study was aimed at evaluating the potential of fungi isolated from garri processing environment for amylase secretion and conduct a time course study of the enzyme production. Fungi were isolated from the soil and waste water from garri processing sites using Standard Microbiological Procedures. The five fungi isolated from garri processing environmental samples were screened using Lugols iodine. Three fungal isolates were then selected on the hydrolysis of starch in qualitative zonation agar plates but one isolate named B among them was better in starch hydrolysis related to highest clear zone plates. After qualitative screening, the three hyper production amylase extracted fungi were identified according to the morphological characteristics. The hyper producing amylase isolate B was then identified as Aspergillus flavus (RCBBR_AEAFUN2) and recorded as a novel strain in southern Nigeria according to molecular characteristics, which was selected for further studies. The current results of time course study showed that the maximum growth (2.453 g) occurred after 24 h of incubation at pH 4.5 while maximum enzyme production (2.3 U/ml/min) was obtained after 96 h of incubation at pH 3.5. Therefore, the hyper producing amylase from garri processing sites, Aspergillus flavus (RCBBR_AEAFUN2) novel strain has great prospects for future biotechnological and industrial applications and help in ameliorating the environmental pollution posed by the waste-water.

Caseinase Production and Media Optimization from Bacillus subtilis

Caseinase is involved in the breakdown of milk protein casein and converts casein into smaller simple sugars which can be easily utilized by the body for the production of ATP and Fat. Casein can be an instant energy source to the body and involves in muscle building. Caseinase enzyme can be extensively used at the industrial scale for Milk, Textile, Dairy, Paper industry and several other medical purposes. In view of the importance of caseinase, the current research deals with the isolation and identification of caseinase producing bacteria from soil. This is followed by the production of enzyme and its purification. The study also includes its kinetic characterization using the parameters Temperature, pH as well as Carbon and Nitrogen Sources. The organism which was isolated from soil and capable of producing the caseinase enzyme was identified to be Bacillus subtilis based on the Biochemical tests and 16S rRNA sequencing result. The optimal carbon and nitrogen sources were identified to be Glucose and casein respectively. Regarding the optimal conditions, the suitable temperature for maximum enzyme production was found to be 40 0C and pH was 9. When the organism was cultured under the optimal condition using casein as a nitrogen source and glucose as the carbon source, at 40 0C and pH 9, 1590 ng/mL of enzyme production was estimated.

RESPONSE SURFACE METHODOLOGY BASED MEDIUM OPTIMIZATION FOR IMPROVEMENT OF CHOLESTEROL OXIDASE ENZYME PRODUCTION BY NEWLY ISOLATED STREPTOMYCES SP.

Cholesterol oxidase is an important enzyme with great commercial value in many clinical and industrial processes, such as determination of cholesterol levels in food, serum and other clinical samples as well as its potential application, as a biocatalyst for bioconversion of sterol compounds and as an insecticide. A Preliminary comparative study of cholesterol oxidase enzyme production from Streptomyces sp. strain NHIA_CH5 using six different media containing cholesterol in submerged fermentation was investigated. An increase in the production was detected after two days in all of the studied media with a maximum production of 23.7 U/mL, noticed in medium containing soluble starch, glucose, peptone, beef extract, MgSO4.7H2O, and NaCl. A classical (one-variable-at-a-time) method revealed that the elimination of MgSO4.7H2O and NaCl favored cholesterol oxidase enzyme production. Statistical optimization of the other components was studied using the screening method Plackett-Burman design followed by the modeling method of the central composite design (CCD). The results showed that beef extract and glucose are the variables that significantly influenced and considered as the most important factors for cholesterol oxidase production. Experiments of CCD based on response surface methodology determine the optimum concentrations of these significant medium components and revealed maximum cholesterol oxidase enzyme production (50.5 U/mL) at beef extract 55 g/L and glucose 40 g/L which represented 2- fold higher titers than acquired from the non-optimized medium.

Optimization of culture parameters for α-glucosidase production from suspension culture of moss Hyophilla nymaniana (Fleish.) Menzel

BackgroundBryophytes, comprising of the second largest group in the plant kingdom, has attracted a great deal of attention in recent years due to its immense potential to produce biopharmaceuticals. But studies conducted to better understand their chemical composition are limited and scattered. In the present investigation, sequential optimization strategy, based on statistical experimental designs, was employed to enhance the production of α-glucosidase enzyme from moss Hyophilla nymaniana (Fleish.) Menzel by using Taguchi methodology. L16 orthogonal array and five physical parameters including sugar, temperature, pH, rpm, nitrogen source were considered as key parameters for enzyme production. The optimal level of each parameter for maximum glucosidase production by the moss was determined. Analysis of variance (ANOVA) was performed to evaluate statistically significant process factors. ResultsBased on statistical analysis (ANOVA), the optimal combinations of the major constituents of media for maximal α-glucosidase production were evaluated as follows: dextrose 2% contributed maximum on α-glucosidase production followed by ammonium nitrate 1.5%, temperature 24 °C, pH 5.6, and RPM 120. Predicted results showed an enhanced glucosidase (53%) than the basal production medium. ConclusionThe present study highlighted that Taguchi design of experiments approach is better than the conventional optimization technique to determine the optimum level of each of the significant parameters that brings maximum enzyme production.

A new uricase from Bacillus cereus SKIII: Characterization, gene identification and genetic improvement

Twenty-five microbial isolates were investigated for uricase production on uric acid medium. All isolates were obtained from Jeddah, Saudi Arabia. The highest uricase producer was identified as Bacillus cereus SKIII. Using glucose peptone broth at pH 7.5, incubation temperature 30 °C for 3 days with shaking of 150 rpm were the best conditions for maximum enzyme production. Glucose and peptone were the best carbon and nitrogen sources. The molecular weight of the purified enzyme was34.5 KDa, and isoelectric point was 7.9. The optimum pH and temperature were pH 8.0 and 35 °C, respectively. It was stable at 35 °C for 60 min, but thermally inactivated at 60 °C after 60 min Its enzymatic activity was enhanced by Mg2+, Ca2+,Fe2+, Mn2+, Zn2+ions and inhibited by Co2+, Na+, Hg2+, Ag+ ions and EDTA at 1 mM. Uricase production was enhanced using UV mutation and the obtained mutant produced six times higher than the original isolate. An amplicon 900 bp of uricase gene (Pucl) was sequenced (accession number MF417635). No remarkable difference was noticed in B. cereus SKIII and SKm mutant nucleotide sequences. In conclusion, SKIII and SKm are promising strains in uricase production for biotechnological applications.

Statistical optimization of amylase production and its purification from a palm wine isolate Bacillus sp., Q-164

Abstract Adaptation of eco-friendly fermentation methods not only reduce environmental pollution and also reduce the cost of large scale production of enzymes. In the present study, the most significant amylase producing strain was isolated from palm wine and identified as Bacillus sp., Q-164 by the molecular 16SrRNA sequence technique. The bacterium exhibited maximum enzyme production in 3 days of the time interval, at neutral pH with 1% of lotus and pineapple stem residues. Enzyme yield further improved to 126.17 ± 0.2 U/mL and 113.4 ± 0.26 U/mL by acid hydrolyzed lotus and pineapple stems respectively. Three factors were selected to optimize amylase production statistically by Response Surface Methodology (RSM). The most significant factors that promote maximum enzyme production were ATLS, temperature, and peptone concentration (P

Bio removal of proteins, lipids and mucopolysaccharides in tannery hyper saline wastewater using halophilic bacteria

The removal of macro bio-molecules such as proteins, lipids and mucopolysaccharides from tannery hyper saline wastewater was exclusively investigated in this present investigation. The halophilic organisms such as Lysinibacillus fusiformis sp. and Providencia stuatrti sp. were screened and isolated for the production of mixed enzyme (proteolytic, lipolytic and amylolytic nature) which has the tendency to degrade the organics at saline medium (TDS > 6 % w/w). The animal fleshing (ANFL), one of the solid wastes generated in leather industry was used as substrate to produce mixed enzyme. The parameters such as incubation time, reaction temperature, pH, salinity and the substrate concentration were optimized for maximum enzyme production. The purified mixed enzyme produced from Lysinibacillus fusiformis sp. has protease activity, 180 U/mL; lipase activity, 1390 U/mL and amylase activity, 1340 U/mL respectively. The percentage degradation of proteins, lipids and mucopolysaccharides in MLCR were observed to be 85 %, 78 % and 83 % respectively for the treatment of soak liquor wastewater.

Application of Plackett Burman and Box Behnken Design for the Optimization of a-glucosidase Production by Thermophillic Thermomyces dupontii

A consecutive optimization based on statistical approach was applied for a-glucosidase production by both wild and mutant T. dupontii. Plackett Burman design (PBD) with two levels was employed in order to screen the significant effect of different nutritional and physical parameters through submerged fermentation. Among all nine variables tested in PBD, incubation time, inoculum size and ammonium sulphate concentration were selected. The Box-Behnken approach was further applied for process optimization. The a-glucosidase production for both wild and mutant T.dupontii was obtained at 72 h of incubation, 1.25 mL inoculum size and 0.25% ammonium sulphate concentration with relatively 95% correlation between the experimentally predicted and observed values. The duration of maximum enzyme production in RSM was cost-saving and fast. The quadratic model was in satisfactory adjustment with the experimental data with high R2 value which describes 98.90% of response variability of the model. Moreover, the novel approach of this present work is that, consecutive optimization were applied for maximum a-glucosidase production using response surface methodology by both wild and mutant thermophillic T. dupontii. Results revealed that thermophillic mutant T. dupontii could be potential candidate for industrial applications.