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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

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논문

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

학술지

AMB Express

저자명

Kumar, Bikash; Bhardwaj, Nisha; Alam, Ansar; Agrawal, Komal; Prasad, Himanshu; Verma, Pradeep

초록

<P>A cellulase producing fungus <I>Schizophyllum commune</I> NAIMCC-F-03379 was isolated from decomposed leaf sample of <I>Lantana camera.</I> 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 <I>S. commune</I> NAIMCC-F-03379. The optimized medium contained: 1% (w/v) wheat bran, 0.3&nbsp;g/L MgSO<SUB>4,</SUB> 0.8&#x2013;1.0&nbsp;g/L CaCl<SUB>2</SUB>, optimum temperature and pH were 25&nbsp;°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/MnSO<SUB>4</SUB> system showed maximum purification with 10.4-fold increase in activity, 79.5% yield and 0.5 partition coefficient. The cellulase enzyme obtained from <I>S. commune</I> NAIMCC-F-03379 has shown high stability i.e. more than 55% relative activity after 12&nbsp;h of incubation over wide range of temperature (25&#x2013;65&nbsp;°C) and pH (3&#x2013;10). The molecular weight of the cellulase enzyme was estimated as ~ 60&nbsp;kDa by using sodium dodecyl sulphate-polyacrylamide electrophoresis (SDS-PAGE) and zymography. K<SUB>m</SUB> and V<SUB>max</SUB> value of cellulase on carboxy-methyl cellulose were obtained as 0.0909&nbsp;mg/mL and 45.45&nbsp;&mu;mol/min&nbsp;mg respectively. Rice straw and wheat bran were subjected to hydrolysis using cellulase and cellulase&#x2013;xylanase cocktail and analysed by thin layer chromatography and high performance liquid chromatography (HPLC). The HPLC analysis showed glucose concentration of 1.162&nbsp;mg/mL after enzymatic hydrolysis of rice straw.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (10.1186/s13568-018-0696-y) contains supplementary material, which is available to authorized users.</P>

발행연도

2018

발행기관

Springer Berlin Heidelberg

라이선스

cc-by

ISSN

2191-0855

8

1

페이지

pp.173

주제어

Schizophyllum commune; Cellulase; Response surface methodology; Aqueous two-phase system; Hydrolysis

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1 2023-12-11

논문; 2018-10-17

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