Cargando…
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
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 surfac...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6192944/ https://www.ncbi.nlm.nih.gov/pubmed/30334113 http://dx.doi.org/10.1186/s13568-018-0696-y |
_version_ | 1783363981514113024 |
---|---|
author | Kumar, Bikash Bhardwaj, Nisha Alam, Ansar Agrawal, Komal Prasad, Himanshu Verma, Pradeep |
author_facet | Kumar, Bikash Bhardwaj, Nisha Alam, Ansar Agrawal, Komal Prasad, Himanshu Verma, Pradeep |
author_sort | Kumar, Bikash |
collection | PubMed |
description | 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 MgSO(4,) 0.8–1.0 g/L CaCl(2), 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/MnSO(4) 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. K(m) and V(max) 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 MATERIAL: The online version of this article (10.1186/s13568-018-0696-y) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6192944 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-61929442018-10-18 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 Kumar, Bikash Bhardwaj, Nisha Alam, Ansar Agrawal, Komal Prasad, Himanshu Verma, Pradeep AMB Express Original Article 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 MgSO(4,) 0.8–1.0 g/L CaCl(2), 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/MnSO(4) 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. K(m) and V(max) 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 MATERIAL: The online version of this article (10.1186/s13568-018-0696-y) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2018-10-17 /pmc/articles/PMC6192944/ /pubmed/30334113 http://dx.doi.org/10.1186/s13568-018-0696-y Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article Kumar, Bikash Bhardwaj, Nisha Alam, Ansar Agrawal, Komal Prasad, Himanshu Verma, Pradeep 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 |
title | 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 |
title_full | 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 |
title_fullStr | 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 |
title_full_unstemmed | 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 |
title_short | 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 |
title_sort | 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 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6192944/ https://www.ncbi.nlm.nih.gov/pubmed/30334113 http://dx.doi.org/10.1186/s13568-018-0696-y |
work_keys_str_mv | AT kumarbikash productionpurificationandcharacterizationofanacidalkaliandthermotolerantcellulasefromschizophyllumcommunenaimccf03379anditsapplicationinhydrolysisoflignocellulosicwastes AT bhardwajnisha productionpurificationandcharacterizationofanacidalkaliandthermotolerantcellulasefromschizophyllumcommunenaimccf03379anditsapplicationinhydrolysisoflignocellulosicwastes AT alamansar productionpurificationandcharacterizationofanacidalkaliandthermotolerantcellulasefromschizophyllumcommunenaimccf03379anditsapplicationinhydrolysisoflignocellulosicwastes AT agrawalkomal productionpurificationandcharacterizationofanacidalkaliandthermotolerantcellulasefromschizophyllumcommunenaimccf03379anditsapplicationinhydrolysisoflignocellulosicwastes AT prasadhimanshu productionpurificationandcharacterizationofanacidalkaliandthermotolerantcellulasefromschizophyllumcommunenaimccf03379anditsapplicationinhydrolysisoflignocellulosicwastes AT vermapradeep productionpurificationandcharacterizationofanacidalkaliandthermotolerantcellulasefromschizophyllumcommunenaimccf03379anditsapplicationinhydrolysisoflignocellulosicwastes |