Heterologous protein secretion in Lactococcus lactis is enhanced by the Bacillus subtilis chaperone-like protein PrsA.
The Bacillus subtilis lipoprotein PrsA enhances the yield of several homologous and heterologous exported proteins in B. subtilis by being involved in the posttranslocational stage of the secretion process. In this work, we have studied the effect of B. subtilis PrsA on the secretion of Bacillus amyloliquefaciens alpha-amylase (AmyQ), a target protein for PrsA, and Bacillus licheniformis penicillinase (PenP) a nontarget protein for PrsA, in Lactococcus lactis. Two compatible plasmids were constructed and introduced into L. lactis strain NZ9000: one high copy plasmid, expressing the AmyQ gene (amyQ) or the PenP gene (penP), and one low copy plasmid, expressing the PrsA encoding gene (prsA). When amyQ and prsA were simultaneously expressed under the nisin-inducible promoter P( nisA ), Western blotting experiments revealed a 15- to 20-fold increase in the total yield of AmyQ and a sixfold increase in secreted AmyQ activity, compared to a control strain lacking prsA. When expressed under the same induction conditions, PrsA had no effect on the secretion or total yield of PenP. These results show that the secretion yield of some heterologous proteins can be significantly increased in L. lactis when coproduced with the B. subtilis PrsA protein.
Lindholm
Agneta
A
Division of Microbiology and Epidemiology, Department of Basic Veterinary Sciences, University of Helsinki, Helsinki 00014, Finland.
Ellmén
Ulla
U
Tolonen-Martikainen
Marja
M
Palva
Airi
A
D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology*; D015153:Blotting, Western; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D008074:Lipoproteins / Q000235:genetics* / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology*; D010405:Penicillinase / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014169:Transformation, Bacterial; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; C082160:prsA protein, bacteria; D000516:alpha-Amylases; D010405:Penicillinase