生物加工过程Issue(6)：44-51,8.DOI:10.3969/j.issn.1672-3678.2014.06.009
脯氨酸4羟化酶在大肠杆菌中的密码子优化表达及对反式4羟脯氨酸生物合成的作用
Expression of codon-optimized proline-4-hydroxylase in Escherichia coli and its effect on trans-4-hydroxyproline biosynthesis
摘要
Abstract
To improve expression of proline 4⁃hydroxylase in Escherichia coli,the DNA sequence encoding proline 4⁃hydroxylase was adjusted based on the codon bias of E�coli and the mRNA secondary structure�The optimized proline⁃4⁃hydroxylase gene was overexpressed in E�coli BL21 ( DE3 ) under a tryptophan tandem promoter in pUC19 vector�In shaking flask culture, trans⁃4⁃hydroxyproline was accumulated to ( 0�492 ± 0�034 ) g/L in 8 h with L⁃proline as substrate�For an enhanced trans⁃4⁃hydroxyproline production by E�coli BL21 ( DE3 ) , the effect of constant feeding strategy on trans⁃4⁃hydroxyproline production in a fermentor was studied�With constant feeding rate at 18 g/h,a maximum trans⁃4⁃hydroxyproline titer of 42�5 g/L with a high trans⁃4⁃hydroxyproline productivity (0�966 g/(L·h)) was achieved.关键词
重组大肠杆菌/脯氨酸 4 羟化酶/反式 4 羟脯氨酸/密码子优化/发酵/条件优化Key words
recombinant Escherichia coli/proline-4-hydroxylase/trans-4-hydroxyproline/codon optimization/fermentation/optimization分类
生物科学引用本文复制引用
刘合栋,袁春伟,张震宇..脯氨酸4羟化酶在大肠杆菌中的密码子优化表达及对反式4羟脯氨酸生物合成的作用[J].生物加工过程,2014,(6):44-51,8.基金项目
国家自然科学基金（30800018、30970058）；高等学校博士学科点专项科研基金（200802951036）；工业生物技术教育部重点实验室主任基金（ KLIB ZR200801）；江苏省自然科学基金 （）
