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Applied and Environmental Microbiology, February 2000, p. 588-598, Vol. 66, No. 2
Unité de Génétique
Microbienne1 and Unité d'Ecologie
et de Physiologie du Système
Digestif,2 Institut National de la Recherche
Agronomique, 78352 Jouy en Josas Cedex, France
Received 28 June 1999/Accepted 28 October 1999
The extracellular Staphylococcus hyicus lipase was
expressed under the control of different promoters in Lactococcus
lactis and Bacillus subtilis. Its expression at high
and moderate levels is toxic for the former and the latter hosts,
respectively. In L. lactis, the lipase was expressed at a
high level, up to 30% of the total cellular proteins, under the
control of the inducible promoter PnisA. About 80% of the
lipase remained associated with the cells. Close to half of this amount
remained associated with the inner side of the cytoplasmic membrane as
unprocessed pre-pro-lipase. The other half was trapped by the cell wall
and partially degraded at the N-terminal end. This result suggests that
extracellular proteases degrade the lipase. Surprisingly, the kinetics
and the pattern of lipase degradation were different in the two
L. lactis subspecies, L. lactis subsp.
cremoris and L. lactis subsp.
lactis. The extracellular proteolytic systems that degrade
lipase are thus different in these closely related subspecies. The
incorrect export of the lipase is not due to an inappropriate leader
peptide but may be due to an inefficiency of several steps of lipase
secretion. We propose that (i) the S. hyicus lipase may
require a special accessory system to be correctly exported or (ii) the
kinetics of lipase synthesis may be a critical factor for proper folding.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Expression of the Staphylococcus hyicus Lipase in
Lactococcus lactis
*
Corresponding author. Mailing address: Unité de
Génétique Microbienne, Institut National de la Recherche
Agronomique, 78352 Jouy en Josas Cedex, France. Phone: 33-1 34 65 25 25. Fax: 33-1 34 65 25 21. E-mail:
renault{at}biotec.jouy.inra.fr.
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