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Effects of replacement of promoters and modification of the leader peptide region of the amy gene of Streptomyces griseus on synthesis and secretion of α-amylase by Streptomyces lividans

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Summary

Five different mutations were introduced into the leader peptide region of the α-amylase gene of Streptomyces griseus IMRU 3570. A mutation which increased the positive charge of the N-terminal region of the leader peptide enhanced the secretion of α-amylase by two- to threefold. Replacement of the native promoter of the amylase gene by the promoter of the Tn5 neo gene or by the promoter of the saf gene resulted in a 16-fold increase in α-amylase secretion. The enhanced secretion of α-amylase obtained by using the most efficient promoters was due to a correlated increase in the amount of transcript formed. The translation and secretion processes in S. lividans are not a bottleneck for enzyme secretion even at very high transcription rates, since stimulation of transcription of the α-amylase gene results in a proportionate increase in secretion of the enzyme.

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Authors and Affiliations

  1. Section of Microbiology, Department of Ecology, Genetics and Microbiology, Faculty of Biology, University of León, E-24071, León, Spain

    Tomás Vigal, José A. Gil, Antonio Daza, M. Dolores García-González, Pablo Villadas & Juan F. Martín

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  1. Tomás Vigal
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  2. José A. Gil
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  3. Antonio Daza
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  4. M. Dolores García-González
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  5. Pablo Villadas
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  6. Juan F. Martín
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Communicated by J.W. Lengeler

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Vigal, T., Gil, J.A., Daza, A. et al. Effects of replacement of promoters and modification of the leader peptide region of the amy gene of Streptomyces griseus on synthesis and secretion of α-amylase by Streptomyces lividans . Molec. Gen. Genet. 231, 88–96 (1991). https://doi.org/10.1007/BF00293826

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