TY - JOUR
T1 - Directed evolution and expression tuning of geraniol synthase for efficient geraniol production in escherichia coli
AU - Tashiro, Miki
AU - Fujii, Akira
AU - Kawai-Noma, Shigeko
AU - Saito, Kyoichi
AU - Umeno, Daisuke
N1 - Funding Information:
This work was supported by Grants-in-Aid for Scientific Research on Innovative Areas from MEXT, Japan (JSPS KAKENHI Grant Number 16K21725, 15H04189, 15K14228). D.U. is supported by the PRESTO (JST) and the MEXT/JSPS Grant-in-Aid for Scientific Research on Innovative Areas 23108507. M.T. is supported by a JSPS fellowship for young scientists. This work is financially supported by The Hamaguchi Foundation for the Advancement of Biochemistry, the Futaba Electronics Memorial Foundation, and the Shorai Foundation for Science and Technology.
Publisher Copyright:
© 2017 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation.
PY - 2017
Y1 - 2017
N2 - To achieve an efficient production of geraniol and its derivatives in Escherichia coli, we aimed to improve the activity of geraniol synthase (GES) through a single round of mutagenesis and screening for higher substrate consumption. We isolated GES variants that outperform their parent in geraniol production. The analysis of GES variants indicated that the expression level of GES was the bottleneck for geraniol synthesis. Over-expression of the mutant GESM53 with a 5′-untranslated sequence designed for high translational efficiency, along with the additional expression of mevalonate pathway enzymes, isopentenyl pyrophosphate isomerase, and geranyl pyrophosphate synthase, yielded 300 mg/L/12 h geraniol and its derivatives (>1000 mg/L/42 h in total) in a shaking flask.
AB - To achieve an efficient production of geraniol and its derivatives in Escherichia coli, we aimed to improve the activity of geraniol synthase (GES) through a single round of mutagenesis and screening for higher substrate consumption. We isolated GES variants that outperform their parent in geraniol production. The analysis of GES variants indicated that the expression level of GES was the bottleneck for geraniol synthesis. Over-expression of the mutant GESM53 with a 5′-untranslated sequence designed for high translational efficiency, along with the additional expression of mevalonate pathway enzymes, isopentenyl pyrophosphate isomerase, and geranyl pyrophosphate synthase, yielded 300 mg/L/12 h geraniol and its derivatives (>1000 mg/L/42 h in total) in a shaking flask.
KW - High-throughput screening
KW - Metabolic engineering
KW - Mevalonate pathway
KW - Monoterpene
KW - RBS score
UR - http://www.scopus.com/inward/record.url?scp=85034669902&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85034669902&partnerID=8YFLogxK
U2 - 10.2323/jgam.2017.01.006
DO - 10.2323/jgam.2017.01.006
M3 - Article
C2 - 28954964
AN - SCOPUS:85034669902
SN - 0022-1260
VL - 63
SP - 287
EP - 295
JO - Journal of General and Applied Microbiology
JF - Journal of General and Applied Microbiology
IS - 5
ER -