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Characterization of the $alpha$-Galactosidase Gene from Leuconostoc mesenteroides SY1
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  • Characterization of the $alpha$-Galactosidase Gene from Leuconostoc mesenteroides SY1
저자명
KIM. JONG HWAN,PARK. JAE-YONG,JEONG. SEON-JU,CHUN. JIYEON,LEE. JONG HOON,CHUNGZ. DAE KYUN,KIM. JEONG HWAN
간행물명
Journal of microbiology and biotechnology
권/호정보
2005년|15권 4호|pp.800-808 (9 pages)
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한국미생물생명공학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Leuconostoc mesenteroides SY1, an isolate from kimchi, was able to ferment $alpha$-galactosides, such as melibiose and raffinose. $alpha$-Galactosidase ($alpha$-Gal) activity was higher in cells grown on melibiose and raffinose than cells grown on galactose, sucrose, and fructose. $alpha$-Gal activity was not detected in cells grown on glucose, indicating the operation of carbon catabolite repression (CCR). A 6 kb DNA fragment was PCR amplified using a primer set based on the nucleotide sequence of a putative $alpha$-galactosidase gene (aga) from L. mesenteroides ATCC 8293. Nucleotide sequencing of the 6 kb fragment confirmed the presence of aga and other genes involved in the galactosides utilization, and the gene order was galR (transcriptional regulator)-aga-gaIK (galactokinase)-gaIT (galactose-1-phosphate uridylyltransferase). Northern blotting experiment showed that aga, gaIK, and gaIT constituted the same operon, that the transcription was induced by galactosides, such as melibiose and raffinose, whereas gaIR was independently transcribed as a monocistronic gene, and that the level of transcription was fairly constant. The aga was overexpressed in E. coli BL21 (DE3) using pET26b(+) vector, and $alpha$-Gal was accumulated in E. coli as an inclusion body.