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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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Correspondence to Juan F. Martín.

Additional information

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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Key words

  • Protein secretion
  • Leader peptide
  • In vitro mutagenesis
  • Streptomyces
  • α-Amylase