Title
Expression of Bacillus subtilis levanase gene in Lactobacillus plantarum and Lactobacillus casei
Author
Wanker, E.
Leer, R.J.
Pouwels, P.H.
Schwab, H.
Centraal Instituut voor Voedingsonderzoek TNO
Publication year
1995
Abstract
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 (pE-SIEW2), 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 pE-SIEW2, 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 β-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.
Subject
Nutrition
Bacillus subtilis
Comparative Study
Genes, Bacterial
Glycoside Hydrolases
Inulin
Lactobacillus
Lactobacillus casei
Plasmids
Selection (Genetics)
Support, Non-U.S. Gov't
Transformation, Genetic
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DOI
https://doi.org/10.1007/s002530050406
TNO identifier
51991
ISSN
0175-7598
Source
Applied Microbiology and Biotechnology, 43 (2), 297-303
Document type
article