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

• Published in: Journal of general and applied microbiology Vol. 68; no. 3; pp. 134 - 142
• Main Authors: Tian, Yanjie, Xu, Jia, Shi, Jianing, Kong, Mengyuan, Guo, Changjiang, Cui, Caixia, Wang, Yongtao, Wang, Yan, Zhou, Chenyan
• Format: Journal Article
• Language: English
• Published: Tokyo Applied Microbiology, Molecular and Cellular Biosciences Research Foundation 01.01.2022; Japan Science and Technology Agency
• Subjects: Alteromonas macleodii; Alteromonas macleodii HY35; Amino acid sequence; Amino acids; Cloning; Deoxyribonucleic acid; DNA; E coli; enzymatic properties; Escherichia coli; expression; Gene sequencing; Glycosidases; Glycosyl hydrolase; glycosyl hydrolase family11; Hydrolase; Microorganisms; Molecular weight; Nucleotide sequence; Open reading frames; Peptides; Xylanase
• ISSN: 0022-1260; 1349-8037
• DOI: 10.2323/jgam.2021.10.003

A xylanase gene xynZT-1 from Alteromonas macleodii HY35 was cloned and expressed in Escherichia coli (E. coli). The sequencing results showed that the ORF of xynZT-1 was 831 bp. The xylanase DNA sequence encoded a 29 amino acids (aa) signal peptide and a 247-aa mature peptide. The XynZT-1 has been a calculated molecular weight (MW) of 27.93 kDa, isoelectric point (pI) of 5.11 and the formula C1266H1829N327O384S5. The amino acid sequence of the xynZT-1 had a high similarity with that of glycosyl hydrolase family 11 (GHF11) reported from other microorganisms. The DNA sequence encoding mature peptide was subcloned into pET-28a(+) expression vector. The resulted plasmid pET-28a-xynZT-1 was transformed into E. coli BL21(DE3), and the recombinant strain BL21(DE3)/xynZT-1 was obtained. The optimum temperature and pH of the recombinant XynZT-1 were 45 ℃ and 5.0, respectively.