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Stability of the recombinant plasmid carrying theBacillus amyloliquefaciens α-amylase gene inB. subtilis

  • Applied Microbiology
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Summary

Theα-amylase gene ofBacillus amyloliquefaciens has previously been cloned into pUB110 to give the recombinant plasmid, pKTH10 (Palva 1982. Gene 19:81–87). Strains transformed by this plasmid are promising candidates for industrialα-amylase production. The stability of pKTH10 was determined in variousB. subtilis strains possessing specific alleles which affect the level ofα-amylase secretion.B. subtilis strains carrying pKTH10 were cultivated in starch-containing medium for up to 50 generations without antibiotic selection and then screened for the presence of pKTH10. The plasmid proved stable enough (< 1.0% cured after 50 generations) for industrial batchwise enzyme production in two strains, but in asacU9 strain (thesacU9 mutation increases concominantly the production ofα-amylase levansucrase and proteases) 99.9% of cells had lost pKTH10 after 50 generations, although the parental plasmid (pUB110) was stable in this strain (0.09% cured after 50 generations). The instability of pKTH10 in thesacU9 strain seems somehow to be related to high expression of the clonedα-amylase gene: when grown in a medium restrictingα-amylase production, only 0.53% ofsacU9 cells had lost pKTH10 after 50 generations.

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  1. Research Laboratories of the Finnish State Alcohol Company, Alko Ltd, POB 350, SF-00101, Helsinki, Finland

    Jari Olavi Vehmaanperä & Matti Pellervo Korhola

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  1. Jari Olavi Vehmaanperä
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  2. Matti Pellervo Korhola
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Vehmaanperä, J.O., Korhola, M.P. Stability of the recombinant plasmid carrying theBacillus amyloliquefaciens α-amylase gene inB. subtilis . Appl Microbiol Biotechnol 23, 456–461 (1986). https://doi.org/10.1007/BF02346060

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  • DOI: https://doi.org/10.1007/BF02346060

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