Molecular and General Genetics MGG

, Volume 233, Issue 1–2, pp 42–48 | Cite as

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

  • Christian Hanke
  • Jürgen Hess
  • Günter Schumacher
  • Werner Goebel


A fusion gene (ces-hlyAs) was constructed by ligating the genetic information for the C-terminal 60 amino acids (hlyAs) 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 andhlyAs 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.

Key words

Secretion Recombinant DNA Hemolysin HlyB/H1yD complementation OmpT protease 


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Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Christian Hanke
    • 2
  • Jürgen Hess
    • 1
  • Günter Schumacher
    • 2
  • Werner Goebel
    • 1
  1. 1.Institut für Genetik und MikrobiologieUniversity of WürzburgWürzburgGermany
  2. 2.Abteilung für GenetikPenzbergGermany

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