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Expression of ${\alpha}$-Galactosidase Gene from Leuconostoc mesenteroides SY1 in Lactobacillus brevis 2.14  

Lee, Kang-Wook (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University)
Park, Ji-Yeong (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University)
Park, Jae-Yong (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University)
Chun, Ji-Yeon (Department of Food Science and Technology, Sunchon National University)
Kim, Jeong-Hwan (Institute of Agriculture & Life Science, Gyeongsang National University)
Publication Information
Food Science and Biotechnology / v.17, no.5, 2008 , pp. 1115-1118 More about this Journal
Abstract
${\alpha}$-Galactosidase gene (aga) from Leuconostoc mesenteroides SY1 was expressed in a heterologous host, Lactobacillus brevis 2.14 using an Escherichia coli-Leuconostoc shuttle vector, pSJE. pSJEaga (pSJE carrying aga) was introduced into Lactobacillus brevis 2.14 by electroporation and transformation efficiency was $1.1{\times}10^3$ per ${\mu}g$ DNA. L. brevis transformants (TFs) showed higher ${\alpha}$-galactosidase (${\alpha}$-Gal) activities than cells containing pSJE. Transcription levels of aga in L. brevis 2.14 grown on different carbon sources (1%, w/v) were examined by slot blot analysis. Aga transcript levels and ${\alpha}$-Gal activities were higher in cells grown on melibiose, raffinose, and galactose than cells on glucose, sucrose, and fructose. Western blot result showed that L. brevis 2.14 harboring pSJEaga produced much more ${\alpha}$-Gal when grown on melibiose than on glucose.
Keywords
Leuconostoc ${\alpha}$-galactosidase; heterologous gene expression; Lactobacillus brevis; electroporation; pSJE;
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