1. Cloning, over-expression and characterization of a thermo-tolerant xylanase from Thermotoga thermarum
- Author
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Yingjuan Huang, Hao Shi, Hui Zhong, Yu Zhang, Fei Wang, and Xun Li
- Subjects
Time Factors ,Gene Expression ,Bioengineering ,Peptide ,Xylose ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Open Reading Frames ,chemistry.chemical_compound ,Hydrolysis ,Enzyme Stability ,Escherichia coli ,medicine ,Enzyme kinetics ,Cloning, Molecular ,chemistry.chemical_classification ,Endo-1,4-beta Xylanases ,Bacteria ,Chemistry ,Temperature ,General Medicine ,Hydrogen-Ion Concentration ,Xylan ,Recombinant Proteins ,Amino acid ,Molecular Weight ,Kinetics ,Biochemistry ,Xylanase ,Xylans ,Biotechnology - Abstract
The xyn10B gene, encoding the endo-1,4-β-xylanase Xyn10B from Thermotoga thermarum, was cloned and expressed in Escherichia coli. The ORF of the xyn10B was 1,095 bp and encoded to mature peptide of 344 amino acids with a calculated MW of 40,531 Da. The recombinant xylanase was optimally active at 80 °C, pH 6.0 and retained approx. 60 % of its activity after 2 h at 75 °C. Apparent K m , k cat and k cat /K m values of the xylanase for beechwood xylan were 1.8 mg ml(-1), 520 s(-1) and 289 ml mg(-1) s(-1), respectively. The end products of the hydrolysis of beechwood xylan were mainly oligosaccharides but without xylose after 2 h hydrolysis.
- Published
- 2013