Photobiocatalytic Oxyfunctionalization with High Reaction Rate using a Baeyer–Villiger Monooxygenase from Burkholderia xenovorans in Metabolically Engineered Cyanobacteria - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue ACS Catalysis Année : 2022

Photobiocatalytic Oxyfunctionalization with High Reaction Rate using a Baeyer–Villiger Monooxygenase from Burkholderia xenovorans in Metabolically Engineered Cyanobacteria

Hanna Grimm

Résumé

Baeyer−Villiger monooxygenases (BVMOs) catalyze the oxidation of ketones to lactones under very mild reaction conditions. This enzymatic route is hindered by the requirement of a stoichiometric supply of auxiliary substrates for cofactor recycling and difficulties with supplying the necessary oxygen. The recombinant production of BVMO in cyanobacteria allows the substitution of auxiliary organic cosubstrates with water as an electron donor and the utilization of oxygen generated by photosynthetic water splitting. Herein, we report the identification of a BVMO from Burkholderia xenovorans (BVMO Xeno) that exhibits higher reaction rates in comparison to currently identified BVMOs. We report a 10fold increase in specific activity in comparison to cyclohexanone monooxygenase (CHMO Acineto) in Synechocystis sp. PCC 6803 (25 vs 2.3 U g DCW −1 at an optical density of OD 750 = 10) and an initial rate of 3.7 ± 0.2 mM h −1. While the cells containing CHMO Acineto showed a considerable reduction of cyclohexanone to cyclohexanol, this unwanted side reaction was almost completely suppressed for BVMO Xeno , which was attributed to the much faster lactone formation and a 10-fold lower K M value of BVMO Xeno toward cyclohexanone. Furthermore, the whole-cell catalyst showed outstanding stereoselectivity. These results show that, despite the self-shading of the cells, high specific activities can be obtained at elevated cell densities and even further increased through manipulation of the photosynthetic electron transport chain (PETC). The obtained rates of up to 3.7 mM h −1 underline the usefulness of oxygenic cyanobacteria as a chassis for enzymatic oxidation reactions. The photosynthetic oxygen evolution can contribute to alleviating the highly problematic oxygen mass-transfer limitation of oxygendependent enzymatic processes.
Fichier principal
Vignette du fichier
acscatal.1c04555.pdf (2.1 Mo) Télécharger le fichier
Origine : Publication financée par une institution
Licence : CC BY - Paternité

Dates et versions

hal-03864568 , version 1 (21-11-2022)

Licence

Paternité

Identifiants

Citer

Elif Erdem, Lenny Malihan-Yap, Leen Assil-Companioni, Hanna Grimm, Giovanni Davide Barone, et al.. Photobiocatalytic Oxyfunctionalization with High Reaction Rate using a Baeyer–Villiger Monooxygenase from Burkholderia xenovorans in Metabolically Engineered Cyanobacteria. ACS Catalysis, 2022, 12 (1), pp.66-72. ⟨10.1021/acscatal.1c04555⟩. ⟨hal-03864568⟩
28 Consultations
40 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More