Studies on lipase directed export of Escherichia coli beta-lactamase in Staphylococcus carnosus |
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Authors: | Wolfgang Liebl and Friedrich Götz |
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Institution: | (1) Lehrstuhl für Mikrobiologie, Technische Universität München, Arcisstraße 21, D-8000 München 2, Federal Republic of Germany |
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Abstract: | Summary The lipase (lip) gene of Staphylococcus hyicus was used to study the expression of the Escherichia coli -lactamase (bla) gene in S. carnosus. The bla gene, devoid of its promotor and most of the signal sequence, was fused to the lip structural gene at various positions. A set of 11 secretion vectors (pLL1 to pLL11) was isolated and analysed. All secretion vectors caused -lactamase production and activity in S. carnosus. However, the amount of hybrid proteins secreted was influenced by the length of the NH2-terminal lipase portion. An increased concentration, comparable to that of the native lipase, of secreted lipase/-lactamase hybrid proteins was only found when the lipase portion of the construct comprised more than 101 amino acids of the NH2-terminal region of the lipase preprotein; the proposed lipase signal peptide is 36 amino acids long. If the hybrid proteins constructed contained 101 or less amino acids of the NH2-terminal lipase preprotein, only low amounts of secreted hybrid proteins were detectable and a significant portion of the hybrid proteins and -lactamase activity was found in the cellular fraction. The results indicate that the lipase possesses adjacent to the signal peptide a peptide domain that is essential for the secretion of the lipase/-lactamase hybrid proteins.Abbreviations Cm
chloramphenicol
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bla gene
beta lactamase coding gene of Escherichia coli
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lip gene
lipase-coding gene of Staphylococcus hyicus
- PA
polyacrylamide
- PAGE
PA gel electrophoresis
- SDS
sodium dodecyl sulphate
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indicates plasmid-carrier state |
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Keywords: | Gene expression Lipase gene fusion Protein export Secretion vector Staphylococcus carnosus |
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