Sequence analysis, cloning and over-expression of an endoxylanase from the alkaliphilic Bacillus halodurans

• Published in: Biotechnology letters Vol. 27; no. 8; pp. 545 - 550
• Main Authors: ALEJANDRA MARTINEZ, M, DELGADO, Osvaldo D, BAIGORI, Mario D, SINERIZ, Faustino
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.04.2005; Springer Nature B.V
• Subjects: Bacillus - enzymology; Bacillus - genetics; Bacillus halodurans; Bacillus halodurans MIR32; Bacteria; Bacterial Proteins - genetics; Base Sequence; Binding Sites - genetics; Biological and medical sciences; Biotechnology; Cloning, Molecular; DNA, Bacterial - genetics; E coli; Electrophoresis, Polyacrylamide Gel; Endo-1,4-beta Xylanases - genetics; Engineering and Technology; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Industrial Biotechnology; Industriell bioteknik; Microbiology; Molecular Sequence Data; over-expression; Promoter Regions, Genetic - genetics; purification; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; sequence; Sequence Analysis, DNA; Teknik; xylanase; Purification; Enzyme; Alkalophily; Glycosidases; Gene overexpression; Hydrolases; Endo-1,4-β-xylanase; Xylan endo-1,3-β-xylosidase; xylanase sequence; over-expression; Bacillus halodurans MIR32; O-Glycosidases
• ISSN: 0141-5492; 1573-6776; 1573-6776
• DOI: 10.1007/s10529-005-2879-2

The BhMIR32 xyn11A gene, encoding an extracellular endoxylanase of potential interest in bio-bleaching applications, was amplified from Bacillus halodurans MIR32 genomic DNA. The protein encoded is an endo-1,4-beta-xylanase belonging to family 11 of glycosyl hydrolases. Its nucleotide sequence was analysed and the mature peptide was subcloned into pET22b(+) expression vector. The enzyme was over-expressed in a high density Escherichia coli culture as a soluble and active protein, and purified in a single step by immobilised metal ion affinity chromatography with a specific activity of 3073 IU mg-1.