Expression of alpha-galactosidase gene from Leuconostoc mesenteroides SY1 in Leuconostoc citreum

Abstract

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 ago 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 ago was successfully expressed in L. citreum and its transcription was under the carbon catabolite repression (CCR).