TY - JOUR ID - 3786 TI - An Efficient Procedure for the Production of trans-4-Hydroxy-L-proline Using Recombinantly Expressed Proline Hydroxylase JO - Scientia Iranica JA - SCI LA - en SN - 1026-3098 AU - Chen, Jiaojiao AU - Gu, Dandan AU - Li, Tianyun AU - Ju, Jiansong AU - Xue, Zhangwei AU - Li, Cunhui AU - Yan, Jin AU - Zhang, Jinxiu AU - Wang, Li-an AD - College of Life Sciences, Hebei Normal University, No. 20, Road East of 2nd Ring South, Yuhua District, Shijiazhuang, 050024, China AD - Hebei Brant Pharmaceutical Co., LTD, Mayu Industrial Park, Jinzhou, 052260, China AD - Beijing Hongyisifang Radiation Technology Co., LTD, Beijing, 101113, China Y1 - 2015 PY - 2015 VL - 22 IS - 6 SP - 2350 EP - 2357 KW - proline KW - hydroxy-L-proline KW - proline hydroxylases KW - optimization KW - conversion efficiency KW - truncation DO - N2 - Due to the codon usage and high G+C contentof thetrans-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 the various induction conditions with respect to the enzymatic activity of the engineered strain, including the concentration of the inducer IPTG, the cell density before the inducer was added, the induction temperature and the induction time, the conversion efficiency was more than 97%within 48 h. UR - https://scientiairanica.sharif.edu/article_3786.html L1 - https://scientiairanica.sharif.edu/article_3786_73f15af548506a6e3236da0e135bbb7f.pdf ER -