Expression of Bacillus amyloliquefaciens amylase and Vibrio alginolyticus protease A fusion genes.
Previously we reported [Deane, S. M., Maharaj, R., Robb, F. T. & Woods, D. R. (1987) Journal of General Microbiology 133, 2295-2302] that the production of a Vibrio alginolyticus SDS-resistant alkaline serine protease (Pro A) cloned in Escherichia coli was characterized by a 12 h delay between the synthesis of an inactive precursor and secretion of active Pro A. Replacement of the V. alginolyticus promoter region by the alpha-amylase promoter region from Bacillus amyloliquefaciens resulted in the simultaneous synthesis and secretion of Pro A in E. coli. The V. alginolyticus pro A gene cloned on a shuttle vector did not produce active Pro A in Bacillus subtilis. Although Pro A has a typical Gram-positive signal sequence, it was not functional in B. subtilis. Replacement of the Pro A signal sequence with the alpha-amylase signal sequence resulted in the production of active Pro A in B. subtilis.
Department of Microbiology, University of Cape Town, South Africa.
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D000595:Amino Acid Sequence; D000681:Amylases / Q000235:genetics* / Q000378:metabolism; D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004591:Electrophoresis, Polyacrylamide Gel; D005780:Gelatin; D015964:Gene Expression Regulation, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D014733:Vibrio / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D005780:Gelatin; D000681:Amylases; C022325:aspartic proteinase A; D016282:Aspartic Acid Endopeptidases