谷氨酸棒杆菌Sec分泌型基因高效表达载体的构建
Development of a novel gene expression vector for production of heterologous proteins via the general secretory (Sec) pathway in Corynebacterium glutamicum
谷氨酸棒杆菌是用于蛋白分泌生产理想宿主菌。然而，目前谷氨酸棒杆菌中Sec型分泌表达载体非常少，这限制了其在蛋白生产中的应用。本研究构建了一个新谷氨酸棒杆菌Sec型基因高效表达载体pAU5。pAU5载体使用强启动子tac-M来控制基因的组成型转录，谷氨酸棒杆菌核糖体结合位点（RBS）一致序列来起始蛋白质的翻译，强CgR_2070信号肽序列介导蛋白质的Sec途径分泌。枯草芽孢杆菌α-淀粉酶AmyE作为报告蛋白验证pAU5载体的可用性试验结果表明：在谷氨酸棒杆菌中，具有活性的重组AmyE被高效表达并完全分泌到培养基中。pAU5是一个优秀的谷氨酸棒杆菌Sec分泌型基因表达载体，其在重组蛋白的分泌生产中具有非常广泛的应用前景。
生物工程学分子生物学微生物学
微生物学谷氨酸棒杆菌表达载体pAU5Sec途径蛋白质分泌
贾慧敏,徐大庆.谷氨酸棒杆菌Sec分泌型基因高效表达载体的构建[EB/OL].(2017-06-05)[2025-10-21].http://www.paper.edu.cn/releasepaper/content/201706-76.点此复制
Corynebacterium glutamicum is recognized as a favorable host for the secretory production of heterologous proteins. However, to this date, few secretion-type gene expression vectors in C. glutamicum have been developed, which limits applications of C. glutamicum in secretory production of proteins. In this study, we report the development of a novel efficient general secretory (Sec)-dependent type gene expression vector pAU5 in C. glutamicum. The vector pAU5 employs the strong promoter tac-M to control constitutive gene transcription, the consensus ribosome binding site (RBS) sequence of C. glutamicum to ensure protein translation, and the strong Sec-type cgR_2070 signal sequence to mediate protein secretion in C. glutamicum. The α-amylase AmyE from Bacillus subtilis was used as reporter protein to test the applicability of pAU5 in C. glutamicum. The experimental results showed thatDevelopment of a novel secretion-type gene expression vector for production of heterologous proteins via the general secretory (Sec) pathway in Corynebacterium glutamicum the recombinant AmyE was efficiently expressed and completely secreted into the culture medium in an active form by C. glutamicum, suggesting that pAU5 is an efficient Sec-dependent type gene expression vector in C. glutamicum. The vector pAU5 is highly useful for secretory production of heterologous proteins in C. glutamicum.
MicrobiologyCorynebacterium glutamicumExpression vectorpAU5Sec pathwayProtein secretion
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