Expression of alpha-galactosidase gene from Leuconostoc mesenteroides SY1 in Leuconostoc citreum.
A 2.5 kb aga gene encoding alpha-galactosidase (alpha- Gal) from Leuconostoc mesenteroides SY1 was cloned into pSJE, an E. coli-Leuconostoc shuttle vector. The recombinant plasmid, pSJEaga, was introduced into Leuconostoc citreum KCTC3526 (ATCC49370) by electroporation. Transcription level of aga was the highest in cells grown on raffinose (1%, w/v) followed by cells grown on galactose, melibiose, fructose, glucose, and sucrose. Western blot using antibodies against alpha-Gal showed similar results to slot-blot results and enzyme activity measurements. All the results indicated that the heterologous aga was successfully expressed in L. citreum and its transcription was under the carbon catabolite repression (CCR).
Park
Jae-Yong
JY
Institute of Agriculture and Life Science, Gyeongsang National University,Jinju 660-701, Korea.
Jeong
Seon-Ju
SJ
Lee
Ae Ran
AR
Park
Ji Yeong
JY
Jeong
Woo Ju
WJ
Kim
Jeong Hwan
JH
D015153:Blotting, Western; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018274:Electroporation; D015870:Gene Expression; D007934:Leuconostoc / Q000201:enzymology* / Q000235:genetics; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000235:genetics / Q000502:physiology; D000519:alpha-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
D004269:DNA, Bacterial; D000519:alpha-Galactosidase