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Processing by OmpT of fusion proteins carrying the HlyA transport signal during secretion by theEscherichia coli hemolysin transport system


A fusion gene (ces-hlyA s) was constructed by ligating the genetic information for the C-terminal 60 amino acids (hlyA s) ofEscherichia coli hemolysin (H1yA) to the ces gene for a cholesterol esterase/lipase (CE) from aPseudomonas species. Part (about 30 %) of the expressed fusion protein CE-H1yAs was secreted inE. coli carryinghlyB andhlyD genes. Following the insertion between the reporter gene andhlyA s of a linker sequence that contains the information for potential cleavage sites for the outer membrane protease OmpT, two different fusion proteins (PhoA-H1yAs and CE-HlyAs) were shown to be cleaved by OmpT between the two parts during H1yB/H1yD-mediated secretion. Processed PhoA and CE accumulated in the supernatant. The efficiency of cleavage by OmpT was considerably improved by increasedompT gene dose. It was further shown that OmpT preferentially recognizes potential cleavage sites within the linker sequence.

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Correspondence to Werner Goebel.

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Communicated by E.K.F. Bautz

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Hanke, C., Hess, J., Schumacher, G. et al. Processing by OmpT of fusion proteins carrying the HlyA transport signal during secretion by theEscherichia coli hemolysin transport system. Molec. Gen. Genet. 233, 42–48 (1992). https://doi.org/10.1007/BF00587559

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Key words

  • Secretion
  • Recombinant DNA
  • Hemolysin
  • HlyB/H1yD complementation
  • OmpT protease