The in vivo synthesis of plant sesquiterpenes by Escherichia coli

The in vivo synthesis of plant sesquiterpenes by Escherichia coli Three plant genes encoding (+)‐ै‐cadinene, 5‐epi‐aristolochene, and vetispiradiene cyclases were expressed in Escherichia coli to evaluate the potential of this bacterium to synthesize sesquiterpenes in vivo. Various growth temperatures, carbon sources, and host strains were examined to optimize terpene production. The highest levels of sesquiterpene production occurred when the enzymes were expressed in strain DH5ॅ from the trc promoter (Ptrc) of the high‐copy plasmidpTrc99A in M9 medium supplemented with 0.2% (v/v) glycerol at 30°C for 5‐epi‐aristolochene and vetispiradiene and 37°C for (+)‐ै‐cadinene. The highest concentrations of sesquiterpenes observed were 10.3 μg of (+)‐ै‐cadinene, 0.24 μg of 5‐epi‐aristolochene (measured as (+)‐ै‐cadinene equivalents), and 6.4 μg of vetispiradiene (measured as (+)‐ै‐cadinene equivalents) per liter of culture. These sesquiterpene production levels are >500‐fold lower than carotenoid production, both of which are synthesized from endogenous trans‐farnesyl diphosphate (FDP) in E. coli. Based on these results, we conclude that the limiting factor for sesquiterpene synthesis in E. coli is the poor expression of the cyclase enzyme and not supply of the FDP precursor. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 497–503, 2001. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology and Bioengineering Wiley

The in vivo synthesis of plant sesquiterpenes by Escherichia coli

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
Copyright © 2001 John Wiley & Sons, Inc.
ISSN
0006-3592
eISSN
1097-0290
D.O.I.
10.1002/bit.10037
Publisher site
See Article on Publisher Site

Abstract

Three plant genes encoding (+)‐ै‐cadinene, 5‐epi‐aristolochene, and vetispiradiene cyclases were expressed in Escherichia coli to evaluate the potential of this bacterium to synthesize sesquiterpenes in vivo. Various growth temperatures, carbon sources, and host strains were examined to optimize terpene production. The highest levels of sesquiterpene production occurred when the enzymes were expressed in strain DH5ॅ from the trc promoter (Ptrc) of the high‐copy plasmidpTrc99A in M9 medium supplemented with 0.2% (v/v) glycerol at 30°C for 5‐epi‐aristolochene and vetispiradiene and 37°C for (+)‐ै‐cadinene. The highest concentrations of sesquiterpenes observed were 10.3 μg of (+)‐ै‐cadinene, 0.24 μg of 5‐epi‐aristolochene (measured as (+)‐ै‐cadinene equivalents), and 6.4 μg of vetispiradiene (measured as (+)‐ै‐cadinene equivalents) per liter of culture. These sesquiterpene production levels are >500‐fold lower than carotenoid production, both of which are synthesized from endogenous trans‐farnesyl diphosphate (FDP) in E. coli. Based on these results, we conclude that the limiting factor for sesquiterpene synthesis in E. coli is the poor expression of the cyclase enzyme and not supply of the FDP precursor. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 497–503, 2001.

Journal

Biotechnology and BioengineeringWiley

Published: Dec 5, 2001

Keywords: isoprenoid; sesquiterpene; cyclase; (+)‐ै‐cadinene; 5‐ epi ‐aristolochene; vetispiradiene

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