A 'push-pull-restrain' strategy to improve citronellol production in Saccharomyces cerevisiae
| Autor: | Mingdong Yao, Wen-Hai Xiao, Ying-Jin Yuan, Ying Wang, Guo-Zhen Jiang |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Saccharomyces cerevisiae Proteins Iridoid medicine.drug_class Monoterpene Acyclic Monoterpenes Saccharomyces cerevisiae Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Metabolic engineering 03 medical and health sciences chemistry.chemical_compound Farnesyl diphosphate synthase 010608 biotechnology medicine Overproduction 030304 developmental biology Citronellol 0303 health sciences biology Chemistry Geranyltranstransferase biology.organism_classification Yeast Biochemistry Metabolic Engineering biology.protein Monoterpenes Biotechnology |
| Zdroj: | Metabolic engineering. 66 |
| ISSN: | 1096-7184 |
| Popis: | Microbial production of monoterpenes has attracted increasing attention in recent years. Up to date, there are only few reports on the biosynthesis of the monoterpene alcohol citronellol that is widely used as fragrant and pharmaceutical intermediates. Here, we engineered Saccharomyces cerevisiae by employing a “push-pull-restrain” strategy to improve citronellol production based on the reduction of geraniol. Starting from a engineered geraniol-producing strain, different reductases were investigated and the best performing iridoid synthase from Catharanthus roseus (CrIS) resulted in 285.89 mg/L enantiomerically pure S-citronellol in shake flasks. Geranyl diphosphate (GPP), the most important precursor for monoterpenes, was enhanced by replacing the wild farnesyl diphosphate synthase (Erg20) with the mutant Erg20F96W, increasing the citronellol titer to 406.01 mg/L without negative influence on cell growth. Moreover, we employed synthetic protein scaffolds and protein fusion to colocalize four sequential enzymes to achieve better substrate channeling along with the deletion of an intermediate degradation pathway gene ATF1, which elevated the citronellol titer to 972.02 mg/L with the proportion of 97.8% of total monoterpenes in YPD medium. Finally, the engineered strain with complemented auxotrophic markers produced 8.30 g/L of citronellol by fed-batch fermentation, which was the highest citronellol titer reported to date. The multi-level engineering strategies developed here demonstrate the potential of monoterpenes overproduction in yeast, which can serve as a generally applicable platform for overproduction of other monoterpenes. |
| Databáze: | OpenAIRE |
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