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KMID : 0545119950050050257
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Journal of Microbiology and Biotechnology
1995 Volume.5 No. 5 p.257 ~ p.263
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Expression of Bacillus licheniformis α-amylase Gene in Lactobacillus casei Strains
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Kim Jeong-Hwan
Woo Sung-Hee
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Abstract
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As a first step for developing Lactobacillus strains capable of fermenting starch directly, the α-amylase gene (amyL) from Bacillus licheniformis (Kim et al., 1988. Kor. J. Appl. Microbiol. Bioeng. 16: 369-373) was introduced into Lactobacillus casei strains and the level of α-amylase expression in transformants was examined. 3 kb EcoRI fragments encompassing amyL were subcloned into the suitable lactococcal cloning vectors (pSA3, pMG36e, and pIL2530) and then recombinant plasmids were introduced into E. coli and L. casei strains by electroporation. Only one recombinant plasmid, pIL2530α was able to transform few L. casei strains tested at low efficiencies. The transformation efficiencies with the plasmid into L. casei YIT 9018 and L. casei ATCC 4646 were less than 10^2/㎍ pIL2530α. The level of amylase activities in L. casei was five to ten-fold lower than that in E. coli cells. pIL2530α was stably maintained in Lactobacillus strains in the presence of Em (5㎍/㎖) but without antibiotic selection, it was unstable so more than 95% of cells lost plasmids after a week of daily subculturing.
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