Gene cloning of the maoA gene and overproduction of a soluble monoamine oxidase from Klebsiella aerogenes.
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.
Department of Fermentation Technology, Faculty of Engineering, Hiroshima University, Japan.
H
H
Sugino
K
K
Ishibashi
M
M
Sakaue
M
M
Yamashita
Y
Y
Murooka
D002872:Chromosome Deletion; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular / Q000379:methods; D005798:Genes, Bacterial; D005822:Genetic Vectors; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics*; D008995:Monoamine Oxidase / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping
D007527:Isoenzymes; D011994:Recombinant Proteins; D008995:Monoamine Oxidase