理性改造光脱羧酶并用于反式脂肪酸选择性脱羧反应  

Rational Evolution of a Fatty Acid Photodecarboxylase for Highly Selective Photodecarboxylation of trans Fatty Acid

作　　者：曾森海 鲍雨妍 曹奡 黄小强 Senhai Zeng;Yuyan Bao;Ao Cao;Xiaoqiang Huang(State Key Laboratory of Coordination Chemistry,Chemistry and Biomedicine Innovation Center,School of Chemistry and Chemical Engineering,Nanjing University,Nanjing 210023,China)

机构地区：[1]配位化学国家重点实验室,南京大学化学与生物医药创新研究院,南京大学化学化工学院,南京210023

出　　处：《大学化学》2023年第4期243-252,共10页University Chemistry

基　　金：国家自然科学基金面上项目(22277053);江苏省自然科学基金青年科学基金项目(BK20220760)。

摘　　要：立足光酶催化合成和酶的理性改造等研究前沿,设计了一例光酶催化的反式脂肪酸高选择性脱羧去除实验。具体地,本实验借助分子对接技术,理性设计脂肪酸光脱羧酶Cv FAP的突变体V453E-Cv FAP,并实验验证该突变体能够选择性地催化反油酸的脱羧反应,从而实现顺式脂肪酸的富集。实验涉及培养细胞、诱导酶表达、提取酶液、设计反应以及产物的定性、定量分析等内容,非常适合在化学生物学实验或综合化学实验课程中开设。Photoenzymatic synthesis and rational engineering of enzymes are among the most emerging research areas.Herein,a highly selective photodecarboxylation of trans fatty acid was achieved by rational repurposing of the photodecarboxylase.Specifically,via molecular docking,a mutant of the photodecarboxylase CvFAP,namely V453ECvFAP,was rationaly designed,constructed and subjected to the photoactivated decarboxylation of fatty acids.It turned out that the mutant was highly selective towards elaidic acid,thereby enriching oleic acid.Experimental trainings like cells culturing,protein expression,reaction design,and qualitative and quantitative analysis of products were included.The experiment is easy to conduct and safe for undergraduate chemistry experiment.

关 键 词：酶的理性改造 脂肪酸光脱羧酶Cv FAP 反式脂肪酸 光酶催化 脱羧反应 

分 类 号：G64[文化科学—高等教育学] O6[文化科学—教育学]