Biswanath Bhunia

Decolorization and biodegradation of congo red dye by a novel white rot fungus Alternaria alternata CMERI F6

A novel white rot fungus Alternaria alternata CMERI F6 decolorized 99.99% of 600 mg/L congo red within 48 h in yeast extract-glucose medium at 25 °C, pH 5 and 150 rpm. Physicochemical parameters like carbon and nitrogen sources, temperature, pH and aeration were optimized to develop faster decolorization process. Dye decolorization rate was maximal (20.21 mg/L h) at 25 °C, pH 5, 150 rpm and 800 mg/L dye, giving 78% final decolorization efficiency. Scanning electron microscopy and X-ray Diffraction analysis revealed that the fungus become amorphous after dye adsorption. HPLC and FTIR analysis of the extracted metabolites suggested that the decolorization occurred through biosorption and biodegradation. Thermogravimetric analysis (TGA), differential thermal analysis (DTA) and acid-alkali and 70% ethanol treatment revealed the efficient dye retention capability of the fungus. The foregoing results justify the applicability of the strain in removal of congo red from textile wastewaters and their safe disposal.

Potential use of polyphenol oxidases (PPO) in the bioremediation of phenolic contaminants containing industrial wastewater

The present review emphasizes on the use of Polyphenol oxidase (PPO) enzyme in the bioremediation of phenolic contaminants from industrial wastewater. PPO is a group of enzyme that mainly exists in two forms; tyrosinase (E.C. 1.14.18.1) and laccase (E.C. 1.10.3.1) which are widely distributed among microorganisms, plants and animals. These oxidoreductive enzymes remain effective in a wide range of pH and temperature, particularly if they are immobilized on some carrier or matrices, and they can degrade a wide variety of mono and/or diphenolic compounds. However, high production costs inhibit the widespread use of these enzymes for remediation in industrial scale. Nevertheless, bench studies and field studies have shown enzymatic wastewater treatment to be feasible options for biodegradation of phenols through biological route. Nanomaterials-PPO conjugates have been also applied for removal of phenols which has successfully lower down the drawbacks of enzymatic water treatment. Therefore in this article various approaches and current state of use of PPO in the bioremediation of wastewater, as well as the benefits and disadvantages associated with the use of such enzymes have been overviewed.

Classification, mode of action and production strategy of xylanase and its application for biofuel production from water hyacinth

Xylanases are classified under glycoside hydrolase families which represent one of the largest groups of commercial enzymes. Depolymerizing xylan molecules into monomeric pentose units involves the synergistic action of mainly two key enzymes which are endo-β-xylanase and β-xylosidase. Xylanases are different with respect to their mode of action, substrate specificities, biochemical properties, 3D structure and are widely produced by a spectrum of bacteria and fungi. Currently, large scale production of xylanase can be produced through the application of genetic engineering tool which allow fast identification of novel xylanase genes and their genetic variations makes it an ideal enzymes. Due to depletion of fossil fuel, there is urgent need to find out environment friendly and sustainable energy sources. Therefore, utilisation of cheap lignocellulosic materials along with proper optimisation of process is most important for cost efficient ethanol production. Among, various types of lignocellulosic substances, water hyacinth, a noxious aquatic weed, has been found in many tropical. Therefore, the technological development for biofuel production from water hyacinth is becoming commercially worthwhile. In this review, the classification and mode of action of xylanase including genetic regulation and strategy for robust xylanase production have been critically discussed from recent reports. In addition various strategies for cost effective biofuel production from water hyacinth including chimeric proteins design has also been critically evaluated.

Effect of pH and temperature on stability and kinetics of novel extracellular serine alkaline protease (70 kDa).

A novel extracellular serine protease (70 kDa by SDS-PAGE) was purified and characterized. This enzyme retained more than 93% of its initial activity after preincubation for 30 min at 37 °C in the presence of 25% (v/v) tested organic solvents and showed feather degradation activity. The purified enzyme was deactivated at various combinations of pH and temperature to examine the interactive effect of them on enzyme activity. The deactivation process was modeled as first-order kinetics and the deactivation rate constant (k(d)) was found to be minimum at pH 9 and 37 °C. The kinetic analysis of enzyme over a range of pH values indicated two pK values at 6.21 and at 10.92. The lower pK value was likely due to the catalytic histidine in the free enzyme and higher pK value likely reflected deprotonation of the proline moiety of the substrate but ionization of the active site serine is another possibility. Inhibition kinetic showed that enzyme is serine protease because enzyme was competitively inhibited by antipain and aprotinin as these compounds are known to be competitive inhibitors of serine protease. The organic solvent, thermal and pH tolerances of enzyme suggested that it may have potential for use as a biocatalyst in industry.

Statistical Approach for Optimization of Physiochemical Requirements on Alkaline Protease Production from Bacillus licheniformis NCIM 2042.

The optimization of physiochemical parameters for alkaline protease production using Bacillus licheniformis NCIM 2042 were carried out by Plackett-Burman design and response surface methodology (RSM). The model was validated experimentally and the maximum protease production was found 315.28 U using optimum culture conditions. The protease was purified using ammonium sulphate (60%) precipitation technique. The HPLC analysis of dialyzed sample showed that the retention time is 1.84 min with 73.5% purity. This enzyme retained more than 92% of its initial activity after preincubation for 30 min at 37°C in the presence of 25% v/v DMSO, methanol, ethanol, ACN, 2-propanol, benzene, toluene, and hexane. In addition, partially purified enzyme showed remarkable stability for 60 min at room temperature, in the presence of anionic detergent (Tween-80 and Triton X-100), surfactant (SDS), bleaching agent (sodium perborate and hydrogen peroxide), and anti-redeposition agents (Na2CMC, Na2CO3). Purified enzyme containing 10% w/v PEG 4000 showed better thermal, surfactant, and local detergent stability.

Extracellular alkaline protease from Bacillus licheniformis NCIM‐2042: Improving enzyme activity assay and characterization

Optimization of enzyme assay conditions for alkaline protease from Bacillus licheniformis NCIM‐2042 was carried out by a statistical approach. Four key determinants such as pH, temperature, buffer concentration, and incubation time were optimized by response surface methodology using rotatable central composite design. Maximum enzyme activity was found to be at pH 9.0, temperature 75°C in phosphate buffer (50 mM) when incubated for 10 min. Protease was stable over a broad range of pH 6.0–12.0 and it was stable at 50°C for 1 h. The protease was completely inhibited by PMSF (5 mM), suggesting that the enzyme is a serine alkaline protease. This enzyme had good stability in the presence of H2O2, SDS, Triton X‐100, and retained more than 88% of its initial activity after preincubation for 30 min at room temperature in the presence of 25% v/v DMSO, methanol, ethanol, ACN, and 2‐propanol.

Optimization of culture condition for growth and phenol degradation by Alcaligenes faecalis JF339228 using Taguchi Methodology

Abstract The optimization of five process parameters such as pH, agitation, temperature, inoculum percentage and incubation time were optimized by Taguchi robust design method for obtaining enhanced biomass and phenol degradation by the isolated Alcaligenes faecalis JF339228 from Durgapur steel industry (DSP), India. About 18 experiments were conducted with a different combination of factors and the results obtained in terms of growth of specific bacterial strain and phenol degradation rates were processed in the Qualitek-4 software to study the main effect of individual factors. The main effect, interaction effects and optimal levels of the process factors were determined using signal-to-noise (S/N) ratio. The effect of factors has been studied for bacterial growth and phenol degradation by A. faecalis JF339228. Optimization of the said parameters has been evaluated by Taguchi method and analysed by analysis of variance. Predicted results showed enhanced process performance such as biomass (131.78%) and ...

Optimization of physicochemical parameters for phenol biodegradation by Candida tropicalis PHB5 using Taguchi Methodology

AbstractThe Taguchi orthogonal array (OA) design of experiments methodology, a quality optimization tool, was used to improve the phenol biodegradation potential of the yeast Candida tropicalis PHB5. At three levels, an OA was selected to analyze the effects of the different physicochemical process factors. Experiments were undertaken to confirm the effectiveness of this method and the main factors affecting the growth of C. tropicalis on phenol and its subsequent degradation were found, together with the optimal factor levels. Predicted results showed that biomass yield could be increased from 1,051.96 to 2,495.74 mg/l and the subsequent amount of phenol degraded could be increased from 879.42 to 2,386.43 mg/l. Based on Taguchi methodology, an overall enhancement of growth by 137.24% and phenol degradation by 171.49% could be attained. Validation experiments showed that the growth and phenol biodegradation was significantly improved by up to 132.4% and 165.64%, respectively.

Rapid development of xylanase assay conditions using Taguchi methodology

ABSTRACT The present investigation is mainly concerned with the rapid development of extracellular xylanase assay conditions by using Taguchi methodology. The extracellular xylanase was produced from Aspergillus niger (KP874102.1), a new strain isolated from a soil sample of the Baramura forest, Tripura West, India. Four physical parameters including temperature, pH, buffer concentration and incubation time were considered as key factors for xylanase activity and were optimized using Taguchi robust design methodology for enhanced xylanase activity. The main effect, interaction effects and optimal levels of the process factors were determined using signal-to-noise (S/N) ratio. The Taguchi method recommends the use of S/N ratio to measure quality characteristics. Based on analysis of the S/N ratio, optimal levels of the process factors were determined. Analysis of variance (ANOVA) was performed to evaluate statistically significant process factors. ANOVA results showed that temperature contributed the maximum impact (62.58%) on xylanase activity, followed by pH (22.69%), buffer concentration (9.55%) and incubation time (5.16%). Predicted results showed that enhanced xylanase activity (81.47%) can be achieved with pH 2, temperature 50°C, buffer concentration 50 Mm and incubation time 10 min.

Process engineering studies to investigate the effect of temperature and pH on kinetic parameters of alkaline protease production.

An in depth process engineering study on the effect of temperature and pH on kinetic parameters of alkaline protease production by Bacillus licheniformis NCIM-2042 using starch as substrate has been reported.