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

IF 0.8 4区生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of General and Applied Microbiology Pub Date : 2022-11-10 Epub Date: 2022-08-15 DOI:10.2323/jgam.2021.10.003

Yanjie Tian, Jia Xu, Jianing Shi, Mengyuan Kong, Changjiang Guo, Caixia Cui, Yongtao Wang, Yan Wang, Chenyan Zhou

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引用次数: 0

Abstract

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.

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大肠杆菌中异交单胞菌HY35中ghf11木聚糖酶的克隆、表达和特性研究

克隆了一株macleodii Alteromonas HY35的木聚糖酶基因xynZT-1，并在大肠杆菌中表达。测序结果显示，xynZT-1的ORF为831 bp。木聚糖酶DNA序列编码一个29个氨基酸的信号肽和一个247-aa的成熟肽。XynZT-1的分子量为27.93 kDa，等电点为5.11，分子式为C1266H1829N327O384S5。xynZT-1的氨基酸序列与其他微生物中报道的糖基水解酶家族11 (GHF11)的氨基酸序列高度相似。将编码成熟肽的DNA序列亚克隆到pET-28a(+)表达载体上。将得到的质粒pET-28a-xynZT-1转化到大肠杆菌BL21(DE3)中，得到重组菌株BL21(DE3)/xynZT-1。重组蛋白XynZT-1的最佳温度和pH分别为45℃和5.0。

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来源期刊

Journal of General and Applied Microbiology 生物-生物工程与应用微生物

CiteScore

2.40

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.