Heterologous gene expression of bovine plasmin in Lactococcus lactis |
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Authors: | J Arnau E Hjerl-Hansen H Israelsen |
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Institution: | (1) Biotechnological Institute, Anker Engelunds Vej 1, Building 227, DK-2800 Lyngby, Denmark Tel.: + 45 45 87 66 99, Fax: + 45 45 93 28 88 e-mail: arnau@biobase.dk, DK |
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Abstract: | Heterologous production of bovine plasmin was studied in the industrially relevant bacterium Lactococcus lactis. Two sets of lactococcal gene expression signals were coupled to the region of the plasmin gene coding for the serine protease
domain. When the promoter region of the prtP gene was used, plasmin was detected mainly intracellularly in strain BPL25 by Western blot hybridization. The intracellular
presence of plasmin led to physiological stress. Expression of the plasmin gene driven by the promoter and complete signal
sequence of the lactococcal usp45 gene resulted in efficient plasmin secretion in strain BPL420. Cell lysis was observed in strains producing plasmin fragments
including the catalytic domain, but not in control strains, which only produced a non-catalytic region of plasmin. The plasmin
produced was shown to be biologically active.
Received: 2 December 1996 / Received revision: 17 March 1997 / Accepted: 27 April 1997 |
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