Expression of Bacillus subtilis levanase gene in Lactobacillus plantarum and Lactobacillus casei.
Two Lactobacillus-Escherichia coli shuttle vectors, harbouring the levanase gene from Bacillus subtilis under the control of its own promoter (pLPEW1) or behind the E. coli tac promoter (pESIEW2), were constructed. Lactobacillus plantarum showed the same growth characteristics on selective plates and in liquid media containing inulin, after transformation with either pLPEW1 or pESIEW2. L. plantarum transformed with pLPEW1 could be selected on inulin plates, indicating that levanase expression can be used as a food-grade selection system for Lactobacillus. Lactobacillus casei grew faster in inulin-containing medium than L. plantarum after transformation with pESIEW2, but did not grow when harbouring pLPEW1. Inulin-degrading activities of 90 mU/ml were found in culture medium of L. plantarum containing pLPEW1 or pESIEW2, and of 500 mU/ml in medium of L. casei (pESIEW2). Addition of 1 mM isopropyl beta-D-thiogalactoside to the culture medium had no effect on growth and levanase expression in L. plantarum (pESIEW2) and L. casei (pESIEW2) strains. Levanase produced by L. casei (pESIEW2) has a size of 75 kDa and 72 kDa, corresponding to that of unprocessed and mature B. subtilis levanase, respectively, suggesting that the protein produced is recognized and processed by a signal peptidase.
Wanker
E
E
Institut für Biotechnologie, Arbeitsgruppe Genetik, Graz, Austria.
Leer
R J
RJ
Pouwels
P H
PH
Schwab
H
H
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins; D005798:Genes, Bacterial / Q000235:genetics*; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism; D007444:Inulin / Q000378:metabolism; D007778:Lactobacillus / Q000235:genetics / Q000254:growth & development*; D007780:Lacticaseibacillus casei / Q000235:genetics / Q000254:growth & development; D010957:Plasmids / Q000235:genetics; D012641:Selection, Genetic; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D007444:Inulin; D006026:Glycoside Hydrolases; C021512:levanase