Production, purification and characterization of an acid/alkali and thermo tolerant cellulase from Schizophyllum commune NAIMCC-F-03379 and its application in hydrolysis of lignocellulosic wastes

Abstract

A cellulase producing fungus Schizophyllum commune NAIMCC-F-03379 was isolated from decomposed leaf sample of Lantana camera. The nutritional components (wheat bran, magnesium sulphate and calcium chloride concentrations) and physical parameters (temperature and pH) were optimised by response surface methodology for enhanced cellulase production by S. commune NAIMCC-F-03379. The optimized medium contained: 1% (w/v) wheat bran, 0.3 g/L MgSO4, 0.8–1.0 g/L CaCl2, optimum temperature and pH were 25 °C and 5 respectively. Under optimum condition, 5.35-fold increase in CMCase and 6.62-fold increase in FPase activity was obtained as compared to un-optimized condition. Crude cellulase enzyme was subjected to different purification techniques and comparative evaluation of their efficiency was performed. The aqueous two-phase system using polyethylene glycol 8000/MnSO4 system showed maximum purification with 10.4-fold increase in activity, 79.5% yield and 0.5 partition coefficient. The cellulase enzyme obtained from S. commune NAIMCC-F-03379 has shown high stability i.e. more than 55% relative activity after 12 h of incubation over wide range of temperature (25–65 °C) and pH (3–10). The molecular weight of the cellulase enzyme was estimated as ~ 60 kDa by using sodium dodecyl sulphate-polyacrylamide electrophoresis (SDS-PAGE) and zymography. Km and Vmax value of cellulase on carboxy-methyl cellulose were obtained as 0.0909 mg/mL and 45.45 μmol/min mg respectively. Rice straw and wheat bran were subjected to hydrolysis using cellulase and cellulase–xylanase cocktail and analysed by thin layer chromatography and high performance liquid chromatography (HPLC). The HPLC analysis showed glucose concentration of 1.162 mg/mL after enzymatic hydrolysis of rice straw.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0696-y) contains supplementary material, which is available to authorized users.

A detailed overview of xylanases: an emerging biomolecule for current and future prospective

“…Quantity and quality of the fermentation product vary with different substrates. There are various commercially Hydrolysis of Agroresidues and XOS Production Bhardwaj et al (2017Bhardwaj et al ( , 2019 SmF, submerged fermentation; SSF, solid-state fermentation; XOS, xylooligosaccahrides available substrates, i.e., xylan, carboxymethyl cellulose (CMC), pectin, and starch for, i.e., xylanase, cellulase, pectinase, and amylase, respectively (Barman et al 2015;Bhardwaj et al 2017;Kumar et al 2018a). Due to the high cost of commercial substrates and considering the economic feasibility of the process, scientists are working from past several years to find alternative substrates for the production of these enzymes.…”

Developing a method for real-time visualization of cellulase activity

“…We chose CMC as our surface material since it forms a 3D gel-like structure in an aqueous environment and hence can be used in its pure form to investigate cellulase activity 28 , specifically with a spatial resolution that might be prevented when mixed with other materials 20 . In addition, it is widely used as a reagent for cellulase activity investigation in various applications [29][30][31][32][33][34] . We chose acridine orange as a dye to visualize hydrolysis since it is a cationic dye and therefore is highly soluble in water and compatible with CMC 35 .…”

Analisis Termodinamika Molekul Magnesium Sulphate (MgSO4)

“…Termokimia magnesium sulfat dalam fasa solid dengan ∆ , ∆ ,°, dan pada tekanan 1 atm dan suhu 25°C yang masing-masing mempunyai nilai -1284,9 kJ/mol, -1170,7 kJ/mol, 91,6 J/molK dan 96,48 J/molK. Panas reaksi standar (ΔHr°) MgSO4 bernilai negatif maka reaksi bersifat eksotermis (123)(124)(125) .…”

“…Termokimia magnesium sulfat dalam fasa solid dengan ∆ , ∆ ,°, dan pada tekanan 1 atm dan suhu 25°C yang masing-masing mempunyai nilai -1284,9 kJ/mol, -1170,7 kJ/mol, 91,6 J/molK dan 96,48 J/molK. Panas reaksi standar (ΔHr°) MgSO4 bernilai negatif maka reaksi bersifat eksotermis (123)(124)(125) .Magnesium sulfat secara komersial umumnya digunakan dalam dunia farmasi baik penggunaan secara eksternal maupun internal (126)(127)(128)(129) . Selain itu magnesium sulfat digunakan dalam bahan kosmetik karena secara parsial biasanya dapat digunakan sebagai pencegah kerutan sementara akibat peningkatan kekuatan ionnya (25-27; 130; 131) .…”