SECRETION OF STREPTOCOCCUS BOVIS α-AMYLASE BY CORYNEBACTERIUM GLUTAMICUM USING CSPB PROMOTER AND CSPASIGNAL SEQUENCE
Corynebacterium glutamicum is an important microorganism in the industrial production of amino acids. However, for industrial fermentations, the direct use of soluble starch instead of enzyme-treated starch is difficult, because enzymes that hydrolyze soluble starch are lacked in C. glutamicum. We herein constructed a strain of C. glutamicum synthesizing and secreting α-amylase from Streptococcus bovis 148 (AmyA) for the efficient utilization of soluble starch. For the promoter and signal sequence, we chose the cspB promoter of C. glutamicum and the cspA signal peptide of Corynebacterium ammoniagenes. The fusion protein was successfully expressed and secreted into the culture broth in C. glutamicum by SDS-PAGE and Western blot analysis. The measurement of AmyA activity showed that AmyA in the culture was in an active form. The thin-layer chromatography (TLC) and colorimetric method suggested that soluble starch was hydrolyzed by AmyA and the amount of starch was decreased during fernmentation. Although some high-molecular-weight degradation products remained in the culture broth, AmyA secreted by C. glutamicum effectively utilized soluble starch as carbon and energy substrate for bacterial growth.
Corynebacterium glutamicum Starch degradation α-Amylase protein secretion
Yao Wenjuan Deng Xizozhao Chu Chunli Liu Miao Zheng Pu Sun Zhihao Zhang Yun
College of Biological Science and Technology, China Pharmaceutical University, Nanjing 210009, China Huadong Research Institute for Medicine and Biotechniques, Nanjing 210002, China School of Biotechnology, Jiangnan University, Wuxi 214122, China
国际会议
The 6th International Forum on Post-genome Technologies(6IFPT)(第六届国际后基因组生命科学技术学术论坛)
北京
英文
427-430
2009-09-17（万方平台首次上网日期，不代表论文的发表时间）