| High soluble expression of D-amino acid oxidase in Escherichia coli regulated by a native promoter. | |
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MedLine Citation:
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PMID: 18696261 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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To express high-active soluble D-amino acid oxidase (DAAO), a constitutive plasmid that is regulated by a native hydantoinase promoter (P(Hase)), was constructed. A D-amino acid oxidase gene (dao) was ligated with the P(Hase) and cloned into pGEMKT to constitutively express protein of DAAO without the use of any inducer such as isopropyl beta-D-1-thiogalactopyranoside which is poisonous to the cells and environment. The ribosome binding site region, host strain, and fermentation conditions were optimized to increase the expression level. When cultivated in a 5-m(3) fermenter, the enzyme activity of JM105/pGEMKT-R-DAAO grown at 37 degrees C was found to be 32 U/mL and increase 16-fold over cells of BL21(DE3)/pET-DAAO grown at 28 degrees C. These results indicate the success of our approaches to overproducing DAAO in soluble form in Escherichia coli. |
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Authors:
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Yangqiu Liu; Qiang Li; Hongyu Zhu; Jichu Yang |
Publication Detail:
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Type: Journal Article Date: 2008-08-12 |
Journal Detail:
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Title: Applied biochemistry and biotechnology Volume: 158 ISSN: 1559-0291 ISO Abbreviation: Appl. Biochem. Biotechnol. Publication Date: 2009 Aug |
Date Detail:
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Created Date: 2009-07-08 Completed Date: 2009-09-23 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8208561 Medline TA: Appl Biochem Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 313-22 Citation Subset: IM |
Affiliation:
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Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, Beijing 100084, People's Republic of China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amidohydrolases
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genetics D-Amino-Acid Oxidase / genetics, metabolism* Escherichia coli / enzymology*, genetics*, growth & development Fermentation Gene Expression Regulation, Bacterial* Promoter Regions, Genetic / genetics* |
| Chemical | |
Reg. No./Substance:
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EC 1.4.3.3/D-Amino-Acid Oxidase; EC 3.5.-/Amidohydrolases; EC 3.5.2.2/dihydropyrimidinase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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