高效藤壶赖氨酰氧化酶的体外重组表达体系构建  

Establishment of High-Efficiency Recombinant Barnacle-Derived Lysyl Oxidase Expression System

作　　者：刘伟治[1] 滕潞瑶 张小康 王露露 LIU Wei-Zhi;TENG Lu-Yao;ZHANG Xiao-Kang;WANG Lu-Lu(Key Laboratory of Marine Genetics and Breeding, Ministry of Education, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China)

机构地区：[1]中国海洋大学海洋生命学院,海洋生物遗传学与育种教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2020年第9期120-125,共6页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(31472258)资助。

摘　　要：藤壶(Balanus)是重要的海洋污损生物。研究报道在藤壶胶形成过程中,赖氨酰氧化酶(Lysyl oxidase,LOX)起了至关重要的作用。但迄今未见藤壶来源的LOX的重组表达和性质研究的报道,这已成为理解藤壶胶形成机制的瓶颈。为解决此问题,本研究聚焦海洋污损生物藤壶来源的LOX,通过系统筛选原核表达载体和LOX结构域,首次实现了藤壶来源的赖氨酰氧化酶(BalLOX)的高效可溶性表达,表达产率为0.28 g/L,是文献报道的哺乳动物来源LOX的373倍。研究结果表明重组表达的BalLOX具有较高的生物酶活性。该研究为后续深入理解藤壶胶形成机制及LOX在藤壶粘附中的角色和功能奠定了重要基础。Barnacles are important marine biofoulings.Previous studies demonstrated that lysyl oxidase(LOX)plays a crucial role in the formation of barnacle cement.However,no report on the recombinant expression and properties of the barnacle-derived LOX has been reported until now,which has become the bottleneck in understanding the mechanism of barnacle cement formation.In order to solve this issue,this study focused on the LOX from marine biofouling barnacles.And we achieved the high-soluble expression of active barnacle-derived lysyl oxidase(BalLOX)for the first time by screening different prokaryotic expression vectors and LOX domain.The expression yield is 0.28 g/L,which is 373 times higher than mammalian-derived LOX reported previously.The results show that the recombinant BalLOX has high enzyme activity.This study laid a crucial foundation for further understanding of the mechanism of barnacle cement formation and the role of LOX in barnacle adhesion.

关 键 词：赖氨酰氧化酶 藤壶 重组表达 高效 酶活 

分 类 号：Q55[生物学—生物化学] Q78