An efficient procedure for the production of trans-4-hydroxy-L-proline using recombinantly expressed proline hydroxylase

被引：0

作者：

Chen, J. J. ^{[1
]}

Gu, D. D. ^{[1
]}

Li, T. Y. ^{[1
]}

Ju, J. S. ^{[1
]}

Xue, Zh. W. ^{[2
]}

Li, C. H. ^{[2
]}

Yan, J. ^{[3
]}

Zhang, J. X. ^{[1
,4
]}

Wang, L. A. ^{[1
]}

机构：

[1] Hebei Normal Univ, Coll Life Sci, 20 Rd East 2nd Ring South, Shijiazhuang 050024, Peoples R China

[2] Hebei Brant Pharmaceut Co LTD, Jinzhou 052260, Peoples R China

[3] Beijing Hongyisifang Radiat Technol Co LTD, Beijing 101113, Peoples R China

[4] Nankai Univ, State Key Lab Med Chem Biol, Tianjin 300071, Peoples R China

来源：

SCIENTIA IRANICA | 2015年 / 22卷 / 06期

关键词：

Proline; Hydroxy-L-proline; Proline hydroxylases; Optimization; Conversion efficiency; Truncation; ESCHERICHIA-COLI; STREPTOMYCES-GRISEOVIRIDUS; HEAT-SHOCK; 4-HYDROXYLASE; PURIFICATION; PROTEINS; CLONING; BIOSYNTHESIS;

D O I：

暂无

中图分类号：

T [工业技术];

学科分类号：

08 ;

摘要：

Due to the codon usage and high G+C content of the trans-4-proline-L-hydroxylase gene from the Dactylosporangium sp. strain RH1, the whole gene was optimized and cloned into several vectors for expression. In biotransformations with resting cells, the activity of the enzyme was investigated. The in-house modified plasmid pET-M-3C was found to yield the highest enzymatic activity. Additionally, after the primary fragment screening, the conversion efficiency of fragment 1-257 aa was enhanced from 76.60% to 88.97% compared with the full-length proline 4-hydroxylase within 60 h; we also found that truncation of the gene improved the solubility of the encoded protein. After optimizing various induction conditions with respect to the enzymatic activity of the engineered strain, the conversion efficiency was more than 97% within 48 h. (C) 2015 Sharif University of Technology. All rights reserved.

引用

页码：2350 / 2357

页数：8

共 23 条

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