Engineering cyanobacterial chassis for improved electron supply toward a heterologous ene-reductase

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

Authors

Spasic, J (Author) Other; Oliveira, P (Author) FCUP; Pacheco, C (Author) Other; Kourist, R (Author) Other; Paula Tamagnini (Author) FCUP

Journal

Title: Journal of Biotechnology

Vol. 360

Pages: 152-159

ISSN: 0168-1656

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 1 Citation

Scopus - 7 Citations

Other information

Authenticus ID: P-00X-G8G

DOI: 10.1016/j.jbiotec.2022.11.005

Abstract (EN): Cyanobacteria are noteworthy hosts for industrially relevant redox reactions, owing to a light-driven cofactor recycling system using water as electron donor. Customizing Synechocystis sp. PCC 6803 chassis by redirecting electron flow offers a particularly interesting approach to further improve light-driven biotransformations. Therefore, different chassis expressing the heterologous ene-reductase YqjM (namely Delta hoxYH, Delta flv3, Delta ndhD2 and Delta hoxYH Delta flv3) were generated/evaluated. The results showed the robustness of the chassis, that exhibited growth and oxygen evolution rates similar to Synechocystis wild-type, even when expressing YqjM. By engineering the electron flow, the YqjM light-driven stereoselective reduction of 2-methylmaleimide to 2-methylsuccinimide was significantly enhanced in all chassis. In the best performing chassis (Delta hoxYH, lacking an active bidirectional hydrogenase) a 39 % increase was observed, reaching an in vivo specific activity of 116 U g(DCW)(-)1 and an initial reaction rate of 16.7 mM h(-1). In addition, the presence of the heterologous YqjM mitigated substrate toxicity, and the conversion of 2-methylmaleimide increased oxygen evolution rates, in particular at higher light intensity. In conclusion, this work demonstrates that rational engineering of electron transfer pathways is a valid strategy to increase in vivo specific activities and initial reaction rates in cyanobacterial chassis harboring oxidoreductases.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8