Park Jae-Yong, Jeong Seon-Ju, Lee Ae Ran, Park Ji Yeong, Jeong Woo Ju, Kim Jeong Hwan
Institute of Agriculture and Life Science, Gyeongsang National University,Jinju 660-701, Korea.
J Microbiol Biotechnol. 2007 Dec;17(12):2081-4.
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).
将来自肠系膜明串珠菌SY1的编码α-半乳糖苷酶（α-Gal）的2.5 kb aga基因克隆到大肠杆菌-明串珠菌穿梭载体pSJE中。通过电穿孔将重组质粒pSJEaga导入柠檬明串珠菌KCTC3526（ATCC49370）。aga的转录水平在以棉子糖（1%，w/v）生长的细胞中最高，其次是以半乳糖、蜜二糖、果糖、葡萄糖和蔗糖生长的细胞。使用抗α-Gal抗体的蛋白质免疫印迹显示结果与狭缝印迹结果和酶活性测量结果相似。所有结果表明，异源aga在柠檬明串珠菌中成功表达，其转录受碳分解代谢物阻遏（CCR）调控。
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