Related Experiment Video
Updated: Dec 7, 2025
08:37
Light-Controlled Fermentations for Microbial Chemical and Protein Production
Published on: March 22, 2022
4.5K
Stereospecific linalool production utilizing two-phase cultivation system in Pantoea ananatis
Yasushi Hoshino1, Mika Moriya1, Akiko Matsudaira1
1Research Institute for Bioscience Products & Fine Chemicals, Ajinomoto Co., Inc. 1-1, Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan.
Journal of Biotechnology
|September 27, 2020
Summary
This study engineered Pantoea ananatis to produce stereospecific (S)- and (R)-linalool using specific linalool synthase genes. Optimized production yielded significant quantities of enantiomerically pure linalool, overcoming challenges in natural racemic production.
Area of Science:
- Metabolic Engineering
- Synthetic Biology
- Biotechnology
Background:
- Linalool, a monoterpene alcohol, is widely used for its floral scent in food, beverages, and fragrances.
- Natural linalool is typically a racemic mixture, making enantiomer-specific production challenging.
- Stereospecific linalool is valuable for targeted applications in perfumery and flavor industries.
Purpose of the Study:
- To develop a microbial platform for the efficient production of enantiomerically pure (S)- and (R)-linalool.
- To identify and characterize linalool synthase (LINS) genes from various organisms for stereospecific linalool synthesis.
- To optimize the host strain and fermentation conditions for enhanced linalool yield and purity.
Main Methods:
- Screened 16 linalool synthase (LINS) genes from plants and bacteria in Pantoea ananatis host.
- Co-expressed optimized LINS genes (AaLINS for (S)-linalool, ScLINS for (R)-linalool) with a mutated farnesyl diphosphate synthase (ispA*).
- Employed two-phase cultivation with isopropyl myristate to mitigate linalool volatilization and toxicity, and inactivated membrane-bound glucose dehydrogenase to boost carbon flux.
Main Results:
- Identified Actinidia arguta LINS (AaLINS) for (S)-linalool and Streptomyces clavuligerus LINS (ScLINS) for (R)-linalool, achieving 100% enantiomeric excess.
- Optimized strains produced 5.60 µg/L (S)-linalool and 3.71 µg/L (R)-linalool from 60.0 µg/L glucose.
- Two-phase cultivation and metabolic modifications significantly enhanced linalool production and stability.
Conclusions:
- Engineered Pantoea ananatis successfully produces high-purity (S)- and (R)-linalool using specific LINS genes and optimized fermentation.
- This microbial platform offers a viable alternative to traditional methods for obtaining enantiomerically pure linalool.
- Further metabolic engineering could lead to even higher yields for industrial applications.
