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Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production
Published on: September 20, 2016
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Systematic Engineering to Enhance 8-Hydroxygeraniol Production in Yeast
Herong Wang1, Guozhen Jiang1, Nan Liang1
1Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
Journal of Agricultural and Food Chemistry
|March 1, 2023
Summary
This study enhances microbial production of 8-hydroxygeraniol, a natural insect repellent. Engineering yeast achieved over 1.0 g/L, the highest reported microbial titer, via optimized P450 activity and reduced byproduct formation.
Area of Science:
- Biotechnology
- Synthetic Biology
- Microbial Engineering
Background:
- 8-Hydroxygeraniol is a valuable component of insect pheromones and secretions, with potential as a biological insect repellent.
- Microbial biosynthesis offers a sustainable and green alternative for producing 8-hydroxygeraniol.
- The enzymatic conversion by P450 geraniol-8-hydroxylase (G8H) is a key bottleneck in microbial 8-hydroxygeraniol production.
Purpose of the Study:
- To enhance the microbial production of 8-hydroxygeraniol using engineered *Saccharomyces cerevisiae*.
- To overcome the limitations in geraniol hydroxylation and reduce degradation pathways.
- To achieve industrially relevant titers of 8-hydroxygeraniol through metabolic engineering and fermentation optimization.
Main Methods:
- Optimized the interaction between geraniol-8-hydroxylase (G8H) and cytochrome P450 reductase (CPR).
- Engineered the endoplasmic reticulum and improved nicotinamide adenine dinucleotide phosphate (NADPH) supply.
- Deleted genes encoding enzymes (ADH6 and ARI1) responsible for 8-hydroxygeraniol reduction.
- Implemented a carbon restriction strategy in fed-batch fermentation.
Main Results:
- Increased 8-hydroxygeraniol titer by 2.1-fold (158.1 mg/L) through initial engineering strategies.
- Further elevated production to 238.9 mg/L at the shake flask level after deleting key reductase genes.
- Achieved a record titer exceeding 1.0 g/L in 5.0 L fed-batch fermentation, representing the highest reported microbial yield to date.
Conclusions:
- Successfully enhanced *de novo* biosynthesis of 8-hydroxygeraniol in *Saccharomyces cerevisiae*.
- Demonstrated the effectiveness of integrated P450 engineering strategies in microbes.
- Established a robust microbial platform for sustainable production of 8-hydroxygeraniol.
