Cloning, Expression, and Characterization of a GHF 11 Xylanase from Alteromonas macleodii HY35 in Escherichia col i

[摘要] A xylanase gene xynZT-1 from Alteromonas macleodiiHY35 was cloned and expressed in Escherichia coli(E. coli). The sequencing results showed that theORF of xynZT-1 was 831 bp. The xylanase DNAsequence encoded a 29 amino acids (aa) signal peptide and a 247-aa mature peptide. The XynZT-1 hasbeen a calculated molecular weight (MW) of 27.93kDa, isoelectric point (pI) of 5.11 and the formulaC1266H1829N327O384S5. The amino acid sequence ofthe xynZT-1 had a high similarity with that of glycosyl hydrolase family 11 (GHF11) reported fromother microorganisms. The DNA sequence encodingmature peptide was subcloned into pET-28a(+)expression vector. The resulted plasmid pET-28axynZT-1 was transformed into E. coli BL21(DE3),and the recombinant strain BL21(DE3)/xynZT-1 wasobtained. The optimum temperature and pH of therecombinant XynZT-1 were 45 ℃ and 5.0, respectively.

[学科分类] 微生物学和免疫学

[关键词] Alteromonas macleodii HY35;xylanase;glycosyl hydrolase family11;expression;enzymaticproperties