Influence of a cell-wall-associated protease on production of alpha-amylase by Bacillus subtilis.
AmyL, an extracellular alpha-amylase from Bacillus licheniformis, is resistant to extracellular proteases secreted by Bacillus subtilis during growth. Nevertheless, when AmyL is produced and secreted by B. subtilis, it is subject to considerable cell-associated proteolysis. Cell-wall-bound proteins CWBP52 and CWBP23 are the processed products of the B. subtilis wprA gene. Although no activity has been ascribed to CWBP23, CWBP52 exhibits serine protease activity. Using a strain encoding an inducible wprA gene, we show that a product of wprA, most likely CWBP52, is involved in the posttranslocational stability of AmyL. A construct in which wprA is not expressed exhibits an increased yield of alpha-amylase. The potential role of wprA in protein secretion is discussed, together with implications for the use of B. subtilis and related bacteria as hosts for the secretion of heterologous proteins.
School of Microbiological, Immunological, and Virological Sciences, The Medical School, University of Newcastle upon Tyne, Newcastle upon Tyne, NE2 4HH, United Kingdom.
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Stephenson
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Harwood
D001412:Bacillus subtilis / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins; D015153:Blotting, Western; D002473:Cell Wall / Q000737:chemistry / Q000201:enzymology; D004269:DNA, Bacterial / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic; D000516:alpha-Amylases / Q000096:biosynthesis*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D000516:alpha-Amylases; D012697:Serine Endopeptidases; C104015:WprA protein, Bacillus subtilis