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普鲁兰酶基因在解淀粉芽孢杆菌BF7658菌株中的分泌表达

孙娟娟 沈微 石贵阳 王正祥

工业微生物2012,Vol.42Issue(2):18-22,5.
工业微生物2012,Vol.42Issue(2):18-22,5.DOI:10.3969/j.issn.1001-6678.2012.02.004

普鲁兰酶基因在解淀粉芽孢杆菌BF7658菌株中的分泌表达

Secreted expression of the pullulanase in Bacillus amyloliquefaciens BF7658

孙娟娟 1沈微 2石贵阳 3王正祥3

作者信息

  • 1. 江南大学生物工程学院生物资源与生物能源研究中心,江苏无锡214122
  • 2. 江南大学工业生物技术教育部重点实验室,江苏无锡214122
  • 折叠

摘要

Abstract

According to the sequence of pullulanase gene from Bacillus deramificans (NCBI accession number: BD140672), the gene encoding mature peptide of pullulanase was synthesized and designated BdP. BdP was inserted into the downstream of the DNA fragment encoding signal peptide of pUC980, a secreting expression vector of Bacillus. The constructed plasmid was designated pUC980-BiP. Subsequently, pUC980-BdP was introduced into Bacillus amyloliquefaciens BF7658, an industrial mesophilic amylase producer. Active pullulanase was expressed by recombinant B. amyloliquefaciens. Recombinant pullulanase was secreted into medium, the optimal time of pullulanase production was 48 h and the enzymatic activity was 2.8 U/mL. Its optimum pH was 5.0 and its optimum temperature was 60 ℃. More than 50 % of its initial enzymatic activity was still detectable after incubated at 60 ℃ for 3 h. The enzymatic property showed that the recombinant pullulanase could be used in saccharification process of starch .

关键词

普鲁兰酶/解淀粉芽孢杆菌/Bacillus deramificans

Key words

pullulanase/ Bacillus amyloliquefaciens/ Bacillus deramificans

引用本文复制引用

孙娟娟,沈微,石贵阳,王正祥..普鲁兰酶基因在解淀粉芽孢杆菌BF7658菌株中的分泌表达[J].工业微生物,2012,42(2):18-22,5.

基金项目

国家863高技术研究发展计划(2006AA10Z307,2006AA020204). (2006AA10Z307,2006AA020204)

工业微生物

OACSCDCSTPCD

1001-6678

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