title: Cloning, over-expression and characterization of a thermo-tolerant xylanase from Thermotoga thermarum.
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.
authors.affiliation: Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, College of Chemical Engineering, Nanjing Forestry University, Nanjing, Jiangsu, 210037, China, ilyshihao@163.com.
authors.forename: Hao
authors.initials: H
authors.lastname: Shi
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hui
authors.initials: H
authors.lastname: Zhong
authors.forename: Yingjuan
authors.initials: Y
authors.lastname: Huang
authors.forename: Xun
authors.initials: X
authors.lastname: Li
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
mesh_terms: D001419:Bacteria / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D014990:Xylans / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014990:Xylans; D043364:Endo-1,4-beta Xylanases