Expression and characterization of a GH43 endo-arabinanase from Thermotoga thermarum

BMC Biotechnology

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

[1] Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, Nanjing 210037, China

关键词: Thermotoga thermarum;    Endo-arabinanase;    Arabinose;    Arabinan;   

Others  :  834865
DOI  :  10.1186/1472-6750-14-35

 received in 2013-10-08, accepted in 2014-04-24,  发布年份 2014

【 摘 要 】

Background

Arabinan is an important plant polysaccharide degraded mainly by two hydrolytic enzymes, endo-arabinanase and α-L-arabinofuranosidase. In this study, the characterization and application in arabinan degradation of an endo-arabinanase from Thermotoga thermarum were investigated.

Results

The recombinant endo-arabinanase was expressed in Escherichia coli BL21 (DE3) and purified by heat treatment followed by purification on a nickel affinity column chromatography. The purified endo-arabinanase exhibited optimal activity at pH 6.5 and 75°C and its residual activity retained more than 80% of its initial activity after being incubated at 80°C for 2 h. The results showed that the endo-arabinanase was very effective for arabinan degradation at higher temperature. When linear arabinan was used as the substrate, the apparent K_m and V_max values were determined to be 12.3 ± 0.15 mg ml⁻¹ and 1,052.1 ± 12.7 μmol ml⁻¹ min⁻¹, respectively (at pH 6.5, 75°C), and the calculated k_cat value was 349.3 ± 4.2 s⁻¹.

Conclusions

This work provides a useful endo-arabinanase with high thermostability andcatalytic efficiency, and these characteristics exhibit a great potential for enzymatic conversion of arabinan.

【 授权许可】

   
2014 Shi et al.; licensee BioMed Central Ltd.

【 图 表 】

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