Light-driven hydroxylation of testosterone by Synechocystis sp. PCC 6803 expressing the heterologous CYP450 monooxygenase CYP110D1

• Type: Article in International Scientific Journal
• Date: 2022

Authors

Mascia, F (Author) Other; Pereira, SB (Author) Other; Pacheco, CC (Author) Other; Oliveira, P (Author) FCUP; Solarczek, J (Author) Other; Schallmey, A (Author) Other; Kourist, R (Author) Other; Alphand, V (Author) Other; Paula Tamagnini (Author) FCUP

Journal

Title: Green Chemistry

Vol. 24

Pages: 6156-6167

ISSN: 1463-9262

Publisher: Royal Society of Chemistry

Indexing

ISI Web of Knowledge - 1 Citation

Scopus - 14 Citations

Other information

Authenticus ID: P-00W-V8M

DOI: 10.1039/d1gc04714k

Abstract (EN): The selective hydroxylation of steroids through chemical synthesis is a complex reaction with a high environmental impact. The use of photoautotrophic microorganisms expressing heterologous monooxygenases could overcome this problem by fueling the reaction with electrons and O-2 derived from the light-dependent oxidation of water, occurring during photosynthesis. Here, the light-driven selective hydroxylation of testosterone into 15 beta-hydroxytestosterone was achieved using whole-cells of the unicellular cyanobacterium Synechocystis sp. PCC 6803 expressing the heterologous CYP450 monooxygenase, CYP110D1. Additionally, the reaction conditions including cell density, aeration, and substrate concentration were optimized, leading to a maximum specific activity of 1 U g(CDW)(-1). This value is about 2-fold higher than the one achieved using the model heterotrophic bacterium, E. coli, in which was necessary to express not only CYP110D1 but also its electron partners, and to use glucose as a sacrificial electron donor. Altogether, the results obtained here demonstrate the higher efficiency and sustainability (94% atom economy) of testosterone hydroxylation using our engineered Synechocystis chassis, compared to biocatalysis with heterotrophic microorganisms or chemical synthesis.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 13