Gene Cloning, Expression, and Characterization of a Thermostable Xylanase from Nesterenkonia xinjiangensis CCTCC AA001025

• Published in: Applied biochemistry and biotechnology Vol. 162; no. 4; pp. 953 - 965
• Main Authors: Kui, Hong, Luo, Huiying, Shi, Pengjun, Bai, Yingguo, Yuan, Tiezheng, Wang, Yaru, Yang, Peilong, Dong, Shouliang, Yao, Bin
• Format: Journal Article
• Language: English
• Published: New York New York : Humana Press Inc 01.10.2010; Humana Press Inc; Springer; Springer Nature B.V
• Subjects: Amino Acid Sequence; Amino acids; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biochemistry; Biological and medical sciences; Biotechnology; Chemistry; Chemistry and Materials Science; Cloning; Cloning, Molecular; E coli; Endo-1,4-beta Xylanases - chemistry; Endo-1,4-beta Xylanases - genetics; Endo-1,4-beta Xylanases - metabolism; Enzyme Stability; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression; Genetics; Hot Temperature; Hydrogen-Ion Concentration; Kinetics; Micrococcaceae - chemistry; Micrococcaceae - enzymology; Micrococcaceae - genetics; Molecular Sequence Data; Sequence Alignment; Streptomyces; Thermostability; Xylanase; Glycoside hydrolase (GH) family 11; Enzyme; Nesterenkonia xinjiangensis; Endo-1,4-β-xylanase; Gene expression; Thermal stability; Glycosylases; Characterization; Glycosidases; Hydrolases; Glycoside; Xylan endo-1,3-β-xylosidase; Molecular cloning
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-009-8815-5

An endo-β-1,4-xylanase-encoding gene, xyn11NX, was cloned from Nesterenkonia xinjiangensis CCTCC AA001025 and expressed in Escherichia coli. The gene encoded a 192-amino acid polypeptide and a putative 50-amino acid signal peptide. The deduced amino acid sequence exhibited a high degree of similarity with the xylanases from Streptomyces thermocyaneoviolaceus (68%) and Thermobifida fusca (66%) belonging to glycoside hydrolase family 11. After purification to homogeneity, the recombinant Xyn11NX exhibited optimal activity at pH 7.0 and 55 °C and remained stable at weakly acidic to alkaline pH (pH 5.0-11.0). The enzyme was thermostable, retaining more than 80% of the initial activity after incubation at 60 °C for 1 h and more than 40% of the activity at 90 °C for 15 min. The K m and V max values for oat spelt xylan and birchwood xylan were 16.08 mg ml⁻¹ and 45.66 μmol min⁻¹ mg⁻¹ and 9.22 mg ml⁻¹ and 16.05 μmol min⁻¹ mg⁻¹, respectively. The predominant hydrolysis products were xylobiose and xylotriose when using oat spelt xylan or birchwood xylan as substrate.