FOOD SCIENCE ›› 2020, Vol. 41 ›› Issue (2): 202-207.doi: 10.7506/spkx1002-6630-20181105-056

• Bioengineering • Previous Articles     Next Articles

Metabolic Engineering of Escherichia coli for Production of trans-4-Hydroxy-L-Proline

LI Qiang, HAN Yakun, JIANG Shuai, ZHANG Yue, WU Heyun, XU Qingyang, XIE Xixian   

  1. (National and Local United Engineering Lab of Metabolic Control Fermentation Technology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China)
  • Received:2020-01-20 Revised:2020-01-20 Online:2020-01-25 Published:2020-01-19

Abstract: In this study, we obtained a novel L-proline-4-hydroxylase with high activity from Micromonospora sp. CNB394. Then we successfully constructed a trans-4-hydroxy-L-proline-producing strain named HYP15 by enhancing the L-proline synthesis pathway and introducing the high-activity L-proline-4-hydroxylase into Escherichia coli using clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR-Cas9)-mediated gene editing. The strain could efficiently produce trans-4-hydroxy-L-proline utilizing glucose as the carbon source. HYP15 produced 13.6 g/L trans-4-hydroxy-L-proline after 30 h of fermentation in shake flasks and produced 48.6 g/L trans-4-hydroxy-L-proline with conversion ratio of sugar to acid of 21.6% and average productivity of 1.22 g/(L·h) after 40 h of fed-batch fermentation in a 5-L bioreactor and, therefore, could be promising for industrial application.

Key words: L-proline-4-hydroxylase, trans-4-hydroxy-L-proline, Escherichia coli, CRISPR-Cas9, fermentation

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