A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization

Biotechnology for Biofuels

Hao Shi¹  Yu Zhang¹  Xun Li¹  Yingjuan Huang¹  Liangliang Wang¹  Ye Wang¹  Huaihai Ding¹  Fei Wang¹ 

[1] Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, 213337, Nanjing, China

关键词: Birchwood;    Oat spelt;    Beechwood;    Thermostability;    Xylan;    Xylanase;   

Others  :  798145
DOI  :  10.1186/1754-6834-6-26

 received in 2012-11-14, accepted in 2013-02-13,  发布年份 2013

【 摘 要 】

Background

Xylanase is an important component of hemicellulase enzyme system. Since it plays an important role in the hydrolysis of hemicellulose into xylooligosaccharides (XOs), high thermostable xylanase has been the focus of much recent attention as powerful enzyme as well as in the field of biomass utilization.

Results

A xylanase gene (xyn10A) with 3,474 bp was cloned from the extremely thermophilic bacterium Thermotoga thermarum that encodes a protein containing 1,158 amino acid residues. Based on amino acid sequence homology, hydrophobic cluster and three dimensional structure analyses, it was attested that the xylanase belongs to the glycoside hydrolase (GH) families 10 with five carbohydrate binding domains. When the xylanase gene was cloned and expressed in Escherichia coli BL21 (DE3), the specific enzyme activity of xylanase produced by the recombinant strain was up to 145.8 U mg^-1. The xylanase was optimally active at 95°C, pH 7.0. In addition, it exhibited high thermostability over broad range of pH 4.0-8.5 and temperature 55-90°C upon the addition of 5 mM Ca²⁺. Confirmed by Ion Chromatography System (ICS) analysis, the end products of the hydrolysis of beechwood xylan were xylose, xylobiose, xylotriose, xylotetraose, xylopentaose and xylohexaose.

Conclusions

The xylanase from T. thermarum is one of the hyperthermophilic xylanases that exhibits high thermostability, and thus, is a suitable candidate for generating XOs from cellulosic materials such as agricultural and forestry residues for the uses as prebiotics and precursors for further preparation of furfural and other chemicals.

【 授权许可】

   
2013 Shi et al; licensee BioMed Central Ltd.

【 图 表 】

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