表达元件优化促进重组胶原蛋白在谷氨酸棒杆菌中的表达OA北大核心CSTPCD
Efficient production of recombinant collagen in Corynebacterium glutamicum by expression elements optimization
重组胶原蛋白是一种具有广泛应用潜力的生物材料,近年来引起了生物医学、组织工程等众多领域的关注.胶原蛋白的3股螺旋结构赋予其独特的生物学功能和生物相容性,但也增加了其在微生物系统中表达的复杂性.该研究以细菌胶原蛋白V-B作为模式蛋白,通过对表达元件的优化,促进重组胶原蛋白在谷氨酸棒杆菌中的表达.首先通过启动子筛选和发酵时长优化,获得介导V-B表达的最优启动子tac-R0.接着利用RBS calculator设计核糖体结合位点(ribosomal binding site,RBS)和间隔序列(aligned spacing,AS)的突变文库,得到与tac-R0启动子搭配组合的最优RBS和AS,使V-B的产量提高至514 mg/L.此外,通过多基因表达盒的串联组合策略,将多个目的基因串联,最终V-B的产量较初始水平提高了 8.4倍,达697 mg/L,该研究为重组胶原蛋白的工业化生产提供了基础.
Recombinant collagen is a biopolymer with broad potential applications in various fields,such as biomedicine and tissue engineering.The triple-helix structure of collagen imparts unique biological functions and biocompatibilit but also increases the complexity of its expression in microbial systems.In this study,bacterial collagen V-B was used as a model protein to promote the expression of re-combinant collagen in Corynebacterium glutamicum through optimization of expression elements.Firstly,the most efficient promoter tac-R0 was obtained through promoter screening and fermentation duration optimization.Subsequently,the RBS calculator was used to design a mutation library for ribosome binding sites(RBS)and aligned spacing(AS).The RBS and AS with the highest strength were identified,which increased the yield of V-B to 514 mg/L.Furthermore,through the tandem combination of multi-gene expression cassettes,the final yield of V-B increased by 8.4-fold compared to the initial level,reaching 697 mg/L.This study lays a foundation for the industrial produc-tion of recombinant collagen.
程逸凡;张萌;许菲
江南大学生物工程学院,工业生物技术教育部重点实验室,江苏 无锡,214122
重组胶原蛋白谷氨酸棒杆菌启动子核糖体结合位点串联表达
recombinant collagenCorynebacterium glutamicumpromoterribosome binding sitesmulti-gene expression cassettes
《食品与发酵工业》 2024 (014)
1-9 / 9
国家自然科学基金(22078129)
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