Gene cloning of the maoA gene and overproduction of a soluble monoamine oxidase from Klebsiella aerogenes |
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Authors: | Hiroyuki Sugino Kaname Ishibashi Masashi Sakaue Mitsuo Yamashita Yoshikatsu Murooka |
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Institution: | (1) Department of Fermentation Technology, Faculty of Engineering, Hiroshima University, Kagamiyama 1, 724 Higashi-Hiroshima, Japan |
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Abstract: | Summary We cloned the structural gene for monoamine oxidase (maoA) from Klebsiella aerogenes into a pKI212 vector in an maoA mutant strain of K. aerogenes. Deletion analysis and complementation tests of the recombinant plasmid showed that the maoA gene was located entirely within a 4.1-kb segment. In an maoA mutant strain harbouring the cloned maoA gene, synthesis of monoamine oxidase was induced by addition of tyramine and related compounds. Transfer of a plasmid containing the maoA gene into a monoamine oxidase-producing strain of K. aerogenes W70 resulted in about a 30- to 40-fold increase in total production of the enzyme. When cells of K. aerogenes carrying the plasmid containing the maoA gene were grown with tyramine, more than 85% of the monoamine oxidase was produced in soluble form, whereas the parent strain W70 produced most monoamine oxidase as the membrane-bound form.
Offprint requests to: Y. Murooka |
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