Integration of a multi-step heterologous pathway in Saccharomyces cerevisiae for the production of abscisic acid
Journal article, 2019

Background: The sesquiterpenoid abscisic acid (ABA) is mostly known for regulating developmental processes and abiotic stress responses in higher plants. Recent studies show that ABA also exhibits a variety of pharmacological activities. Affordable and sustainable production will be required to utilize the compound in agriculture and as a potential pharmaceutical. Saccharomyces cerevisiae is an established workhorse for the biotechnological production of chemicals. In this study, we constructed and characterised an ABA-producing S. cerevisiae strain using the ABA biosynthetic pathway from Botrytis cinerea. Results: Expression of the B. cinerea genes bcaba1, bcaba2, bcaba3 and bcaba4 was sufficient to establish ABA production in the heterologous host. We characterised the ABA-producing strain further by monitoring ABA production over time and, since the pathway contains two cytochrome P450 enzymes, by investigating the effects of overexpressing the native S. cerevisiae or the B. cinerea cytochrome P450 reductase. Both, overexpression of the native or heterologous cytochrome P450 reductase, led to increased ABA titres. We were able to show that ABA production was not affected by precursor or NADPH supply, which suggested that the heterologous enzymes were limiting the flux towards the product. The B. cinerea cytochrome P450 monooxygenases BcABA1 and BcABA2 were identified as pathway bottlenecks and balancing the expression levels of the pathway enzymes resulted in 4.1-fold increased ABA titres while reducing by-product formation. Conclusion: This work represents the first step towards a heterologous ABA cell factory for the commercially relevant sesquiterpenoid. © 2019 The Author(s).

Metabolic engineering

Synthetic biology

Plant hormone

Abscisic acid

Sesquiterpenoids

Botrytis cinerea

Saccharomyces cerevisiae

Terpenoids

Author

Maximilian Otto

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Paulo Teixeira

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Maria Vizcaino

CMSI

Florian David

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Verena Siewers

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Microbial Cell Factories

1475-2859 (ISSN)

Vol. 18 1 205

Subject Categories

Biochemistry and Molecular Biology

Microbiology

Plant Biotechnology

DOI

10.1186/s12934-019-1257-z

PubMed

31767000

More information

Latest update

1/9/2020 2