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Title: Metabolic engineering of Escherichia coli for the biosynthesis of 2-pyrrolidone

2-Pyrrolidone is a valuable bulk chemical with myriad applications as a solvent, polymer precursor and active pharmaceutical intermediate. A novel 2-pyrrolidone synthase, ORF27, from Streptomyces aizunensis was identified to catalyze the ring closing dehydration of γ-aminobutyrate. ORF27's tendency to aggregate was resolved by expression at low temperature and fusion to the maltose binding protein (MBP). Recombinant Escherichia coli was metabolically engineered for the production of 2-pyrrolidone from glutamate by expressing both the genes encoding GadB, a glutamate decarboxylase, and ORF27. Incorporation of a GadB mutant lacking H465 and T466, GadB_δHT, improved the efficiency of one-pot 2-pyrrolidone biosynthesis in vivo. When the recombinant E. coli strain expressing the E. coli GadB_δHT mutant and the ORF27-MBP fusion was cultured in ZYM-5052 medium containing 9. g/L of l-glutamate, 7.7. g/L of l-glutamate was converted to 1.1. g/L of 2-pyrrolidone within 31. h, achieving 25% molar yield from the consumed substrate.
Authors:
 [1] ;  [2] ;  [2] ;  [2] ;  [2] ;  [3]
  1. Univ. of California, Berkeley, CA (United States); Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States). Synthetic Biology Engineering Research Center
  2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  3. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States). Synthetic Biology Engineering Research Center, Dept. of Chemical and Biomolecular Engineering, California Inst. for Quantitative Biosciences; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division
Publication Date:
Grant/Contract Number:
AC02-05CH11231; 0540879
Type:
Published Article
Journal Name:
Metabolic Engineering Communications
Additional Journal Information:
Journal Volume: 3; Journal Issue: C; Journal ID: ISSN 2214-0301
Publisher:
Elsevier
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Science Foundation (NSF)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; 2-pyrrolidone; e.coli; glutamate; metabolic engineering; biosynthesis
OSTI Identifier:
1394663
Alternate Identifier(s):
OSTI ID: 1393091

Zhang, Jingwei, Kao, Emily, Wang, George, Baidoo, Edward E. K., Chen, Matthew, and Keasling, Jay. D.. Metabolic engineering of Escherichia coli for the biosynthesis of 2-pyrrolidone. United States: N. p., Web. doi:10.1016/j.meteno.2015.11.001.
Zhang, Jingwei, Kao, Emily, Wang, George, Baidoo, Edward E. K., Chen, Matthew, & Keasling, Jay. D.. Metabolic engineering of Escherichia coli for the biosynthesis of 2-pyrrolidone. United States. doi:10.1016/j.meteno.2015.11.001.
Zhang, Jingwei, Kao, Emily, Wang, George, Baidoo, Edward E. K., Chen, Matthew, and Keasling, Jay. D.. 2016. "Metabolic engineering of Escherichia coli for the biosynthesis of 2-pyrrolidone". United States. doi:10.1016/j.meteno.2015.11.001.
@article{osti_1394663,
title = {Metabolic engineering of Escherichia coli for the biosynthesis of 2-pyrrolidone},
author = {Zhang, Jingwei and Kao, Emily and Wang, George and Baidoo, Edward E. K. and Chen, Matthew and Keasling, Jay. D.},
abstractNote = {2-Pyrrolidone is a valuable bulk chemical with myriad applications as a solvent, polymer precursor and active pharmaceutical intermediate. A novel 2-pyrrolidone synthase, ORF27, from Streptomyces aizunensis was identified to catalyze the ring closing dehydration of γ-aminobutyrate. ORF27's tendency to aggregate was resolved by expression at low temperature and fusion to the maltose binding protein (MBP). Recombinant Escherichia coli was metabolically engineered for the production of 2-pyrrolidone from glutamate by expressing both the genes encoding GadB, a glutamate decarboxylase, and ORF27. Incorporation of a GadB mutant lacking H465 and T466, GadB_δHT, improved the efficiency of one-pot 2-pyrrolidone biosynthesis in vivo. When the recombinant E. coli strain expressing the E. coli GadB_δHT mutant and the ORF27-MBP fusion was cultured in ZYM-5052 medium containing 9. g/L of l-glutamate, 7.7. g/L of l-glutamate was converted to 1.1. g/L of 2-pyrrolidone within 31. h, achieving 25% molar yield from the consumed substrate.},
doi = {10.1016/j.meteno.2015.11.001},
journal = {Metabolic Engineering Communications},
number = C,
volume = 3,
place = {United States},
year = {2016},
month = {12}
}