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Studies on lipase directed export of Escherichia coli β-lactamase in Staphylococcus carnosus

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Summary

The lipase (lip) gene of Staphylococcus hyicus was used to study the expression of the Escherichia coli β-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 (pLLβ1 to pLLβ11) was isolated and analysed. All secretion vectors caused β-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/β-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 β-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/β-lactamase hybrid proteins.

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Abbreviations

Cm:

chloramphenicol

bla gene:

beta lactamase coding gene of Escherichia coli

lip gene:

lipase-coding gene of Staphylococcus hyicus

PA:

polyacrylamide

PAGE:

PA gel electrophoresis

SDS:

sodium dodecyl sulphate

[]:

indicates plasmid-carrier state

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Authors and Affiliations

  1. Lehrstuhl für Mikrobiologie, Technische Universität München, Arcisstraße 21, D-8000, München 2, Federal Republic of Germany

    Wolfgang Liebl & Friedrich Götz

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  1. Wolfgang Liebl
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  2. Friedrich Götz
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Communicated by A. Böck

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Liebl, W., Götz, F. Studies on lipase directed export of Escherichia coli β-lactamase in Staphylococcus carnosus . Molec Gen Genet 204, 166–173 (1986). https://doi.org/10.1007/BF00330205

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  • DOI: https://doi.org/10.1007/BF00330205

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