反式-4-羟基-L-脯氨酸羟化酶基因密码子优化及表达  

Improvement of Expression Level of Proline 4-hydroxylase by Codon Optimization

作　　者：张琳琳[1] 张庆[1] 李莉[1] 徐书景[1] 张金秀[1] 鞠建松[1] ZHANG Linlin ZHANG Qing LI Li XU Shujing ZHANG Jinxiu JU Jiansong(College of Life Seienee,Hebei Normal University,Hebei Shijiazhuang 050024,Chin)

机构地区：[1]河北师范大学生命科学学院,河北石家庄050024

出　　处：《河北师范大学学报（自然科学版）》2017年第4期341-347,共7页Journal of Hebei Normal University：Natural Science

基　　金：河北省自然科学基金(C2015205212;C2016205130);病原微生物生物安全国家重点实验室开放基金(SKLPBS1529)

摘　　要：L-羟脯氨酸用途非常广泛,具有较好的应用价值,利用反式-4-羟基-L-脯氨酸羟化酶能将L-脯氨酸通过生物转化法生成反式-4-羟基-L-脯氨酸.对指孢囊菌(Dactylosporangiumsp.)RH1的反式-4-羟基-L-脯氨酸羟化酶基因密码子优化及蛋白可溶性表达条件进行了探索.基因p4hy的GC摩尔百分含量由73.63降至61.45;在23℃下,加入1mmol/L IPTG及分子伴侣pG-KJE8后可溶性蛋白的含量明显增多;密码子优化后的酶蛋白P4HY具有较好的催化生产羟脯氨酸的能力,反应40h后转化率达到72.8%.Trans-4-proline hydroxylase can hydroxylate L-proline to trans-4-hydroxy-L-proline,which is a widely applied in medicine,food,biochemistry and the beauty industry.In order to improve the expression level of proline 4-hydroxylase of Dactylosporangiumsp.RH1,codon optimization and expression condition optimization have been performed.The GC content of gene p4hy was reduced from 73.65 to 61.45.The protein expression level and solubility of P4HY were improved when induced with 1mmol/L ITPG with the existence of chaperone plasmid pG-KJE8 for 6hat 23℃.The codon optimized P4HY has high catalytic activity for conversion of L-proine to trans-4-hydroxy-L-proline,with a conversion rate at 72.8%after 40h＇s reaction.

关 键 词：反式-4-羟基-L-脯氨酸羟化酶 密码子优化 指孢囊菌 羟脯氨酸 

分 类 号：Q815[生物学—生物工程]