{"title":"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","authors":"Yanjie Tian, Jia Xu, Jianing Shi, Mengyuan Kong, Changjiang Guo, Caixia Cui, Yongtao Wang, Yan Wang, Chenyan Zhou","doi":"10.2323/jgam.2021.10.003","DOIUrl":null,"url":null,"abstract":"<p><p>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 C<sub>1266</sub>H<sub>1829</sub>N<sub>327</sub>O<sub>384</sub>S<sub>5</sub>. 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.</p>","PeriodicalId":15842,"journal":{"name":"Journal of General and Applied Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8000,"publicationDate":"2022-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cloning, Expression, and Characterization of a GHF 11 Xylanase from Alteromonas macleodii HY35 in Escherichia coli.\",\"authors\":\"Yanjie Tian, Jia Xu, Jianing Shi, Mengyuan Kong, Changjiang Guo, Caixia Cui, Yongtao Wang, Yan Wang, Chenyan Zhou\",\"doi\":\"10.2323/jgam.2021.10.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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 C<sub>1266</sub>H<sub>1829</sub>N<sub>327</sub>O<sub>384</sub>S<sub>5</sub>. 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.</p>\",\"PeriodicalId\":15842,\"journal\":{\"name\":\"Journal of General and Applied Microbiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.8000,\"publicationDate\":\"2022-11-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of General and Applied Microbiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.2323/jgam.2021.10.003\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/8/15 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q4\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of General and Applied Microbiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.2323/jgam.2021.10.003","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/8/15 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Cloning, Expression, and Characterization of a GHF 11 Xylanase from Alteromonas macleodii HY35 in Escherichia coli.
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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