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利用运动发酵单胞菌中PhoD的天然信号肽构建新型分泌表达系统。

Construction of a novel secretion expression system guided by native signal peptide of PhoD in Zymomonas mobilis.

作者信息

Wu Bo, He Ming-Xiong, Feng Hong, Shui Zong-Xia, Tang Xiao-Yu, Hu Qi-Chun, Zhang Yi-Zheng

机构信息

a College of Life Sciences , Sichuan University , Chengdu , China.

出版信息

Biosci Biotechnol Biochem. 2014;78(4):708-13. doi: 10.1080/09168451.2014.896736. Epub 2014 May 9.

Abstract

In the current study, three native signal peptides (SPs) from PhoC, PhoD, and ZMO0331were investigated and compared to construct novel secretion expression systems in Zymomonas mobilis. The secretion expression of target protein, α-amylase from Bacillus amyloliquefaciens (BAA), guided by PhoD's SP resulted in more hydrolysis of starch than that by the other two SPs. Extracellular and intracellular α-amylase activities of the strain containing PhoD's SP were also higher than the other two strains containing PhoC or ZMO0331's SP. In addition, the evidence by alcohol dehydrogenase activity assay further confirmed that the starch hydrolysis was resulted from the secretion expression of BAA rather than the breakage of cells. Our results indicated that the SP of PhoD is able to serve as a promising candidate to assist secretion expression of heterogeneous genes in Z. mobilis. This will contribute to development of engineered Z. mobilis strains converting starch into ethanol.

摘要

在当前研究中,对来自PhoC、PhoD和ZMO0331的三种天然信号肽(SPs)进行了研究和比较,以构建运动发酵单胞菌中的新型分泌表达系统。由PhoD的信号肽引导的目标蛋白——解淀粉芽孢杆菌α-淀粉酶(BAA)的分泌表达,比其他两种信号肽导致的淀粉水解更多。含有PhoD信号肽的菌株的细胞外和细胞内α-淀粉酶活性也高于含有PhoC或ZMO0331信号肽的其他两种菌株。此外,乙醇脱氢酶活性测定的证据进一步证实,淀粉水解是由BAA的分泌表达引起的,而不是细胞破裂。我们的结果表明,PhoD的信号肽能够作为一个有前景的候选者,来协助运动发酵单胞菌中异源基因的分泌表达。这将有助于开发将淀粉转化为乙醇的工程化运动发酵单胞菌菌株。

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