Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass

Raushan Kumar Singh; Manish Kumar Tiwari; Dongwook Kim; Yun Chan Kang; Priyadharshini Ramachandran; Jung-Kul Lee

Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass

Raushan Kumar Singh, Manish Kumar Tiwari, Dongwook Kim, Yun Chan Kang, Priyadharshini Ramachandran, Jung-Kul Lee^*

^*Corresponding author af dette arbejde

Kemisk Institut

Abstract

An endo-1,4-β-xylanase gene, xylcg, was cloned from Chaetomium globosum and successfully expressed in Escherichia coli. The complete gene of 675 bp was amplified, cloned into the pET 28(a) vector, and expressed. The optimal conditions for the highest activity of the purified recombinant XylCg were observed at a temperature of 40 °C and pH of 5.5. Using oat-spelt xylan, the determined K m, V max, and k cat/K m values were 0.243 mg ml−1, 4,530 U mg−1 protein, and 7,640 ml s−1 mg−1, respectively. A homology model and sequence analysis of XylCg, along with the biochemical properties, confirmed that XylCg belongs to the GH11 family. Rice straw pretreated with XylCg showed 30 % higher conversion yield than the rice straw pretreated with a commercial xylanase. Although xylanases have been characterized from fungal and bacterial sources, C. globosum XylCg is distinguished from other xylanases by its high catalytic efficiency and its effectiveness in the pretreatment of lignocellulosic biomass.

Originalsprog	Engelsk
Tidsskrift	Applied Microbiology and Biotechnology
ISSN	0175-7598
Status	Udgivet - 2012

Citationsformater

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title = "Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass",

abstract = "An endo-1,4-β-xylanase gene, xylcg, was cloned from Chaetomium globosum and successfully expressed in Escherichia coli. The complete gene of 675 bp was amplified, cloned into the pET 28(a) vector, and expressed. The optimal conditions for the highest activity of the purified recombinant XylCg were observed at a temperature of 40 °C and pH of 5.5. Using oat-spelt xylan, the determined K m, V max, and k cat/K m values were 0.243 mg ml−1, 4,530 U mg−1 protein, and 7,640 ml s−1 mg−1, respectively. A homology model and sequence analysis of XylCg, along with the biochemical properties, confirmed that XylCg belongs to the GH11 family. Rice straw pretreated with XylCg showed 30 % higher conversion yield than the rice straw pretreated with a commercial xylanase. Although xylanases have been characterized from fungal and bacterial sources, C. globosum XylCg is distinguished from other xylanases by its high catalytic efficiency and its effectiveness in the pretreatment of lignocellulosic biomass.",

author = "Singh, {Raushan Kumar} and Tiwari, {Manish Kumar} and Dongwook Kim and Kang, {Yun Chan} and Priyadharshini Ramachandran and Jung-Kul Lee",

year = "2012",

language = "English",

journal = "Applied Microbiology and Biotechnology",

issn = "0175-7598",

publisher = "Springer",

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TY - JOUR

T1 - Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass

AU - Singh, Raushan Kumar

AU - Tiwari, Manish Kumar

AU - Kim, Dongwook

AU - Kang, Yun Chan

AU - Ramachandran, Priyadharshini

AU - Lee, Jung-Kul

PY - 2012

Y1 - 2012

N2 - An endo-1,4-β-xylanase gene, xylcg, was cloned from Chaetomium globosum and successfully expressed in Escherichia coli. The complete gene of 675 bp was amplified, cloned into the pET 28(a) vector, and expressed. The optimal conditions for the highest activity of the purified recombinant XylCg were observed at a temperature of 40 °C and pH of 5.5. Using oat-spelt xylan, the determined K m, V max, and k cat/K m values were 0.243 mg ml−1, 4,530 U mg−1 protein, and 7,640 ml s−1 mg−1, respectively. A homology model and sequence analysis of XylCg, along with the biochemical properties, confirmed that XylCg belongs to the GH11 family. Rice straw pretreated with XylCg showed 30 % higher conversion yield than the rice straw pretreated with a commercial xylanase. Although xylanases have been characterized from fungal and bacterial sources, C. globosum XylCg is distinguished from other xylanases by its high catalytic efficiency and its effectiveness in the pretreatment of lignocellulosic biomass.

AB - An endo-1,4-β-xylanase gene, xylcg, was cloned from Chaetomium globosum and successfully expressed in Escherichia coli. The complete gene of 675 bp was amplified, cloned into the pET 28(a) vector, and expressed. The optimal conditions for the highest activity of the purified recombinant XylCg were observed at a temperature of 40 °C and pH of 5.5. Using oat-spelt xylan, the determined K m, V max, and k cat/K m values were 0.243 mg ml−1, 4,530 U mg−1 protein, and 7,640 ml s−1 mg−1, respectively. A homology model and sequence analysis of XylCg, along with the biochemical properties, confirmed that XylCg belongs to the GH11 family. Rice straw pretreated with XylCg showed 30 % higher conversion yield than the rice straw pretreated with a commercial xylanase. Although xylanases have been characterized from fungal and bacterial sources, C. globosum XylCg is distinguished from other xylanases by its high catalytic efficiency and its effectiveness in the pretreatment of lignocellulosic biomass.

M3 - Journal article

SN - 0175-7598

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

ER -

Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass

Abstract

Fingeraftryk

Citationsformater