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Title (Primary) Light‐dependent and aeration‐independent gram‐scale hydroxylation of cyclohexane to cyclohexanol by CYP450 harboring Synechocystis sp. PCC 6803
Author Hoschek, A.; Toepel, J.; Hochkeppel, A.; Karande, R.; Bühler, B.; Schmid, A.;
Journal Biotechnology Journal
Year 2019
Department SOMA;
Volume 14
Issue 8
Language englisch;
POF III (all) R61; T15;
Supplements https://onlinelibrary.wiley.com/action/downloadSupplement?doi=10.1002%2Fbiot.201800724&file=biot201800724-sup-0001-supporting_information_pdf.pdf
Keywords Biocatalysis; Bioprocess development; Cyanobacteria; Oxyfunctionalization; Photobiotechnology
Abstract Oxygenase‐containing cyanobacteria constitute promising whole‐cell biocatalysts for oxyfunctionalization reactions. Photosynthetic water oxidation thereby delivers the required co‐substrates, activated reduction equivalents and O2 sustainably. We developed a recombinant Synechocystis sp. PCC 6803 strain showing unprecedentedly high photosynthesis‐driven oxyfunctionalization activities and evaluated its technical applicability. The cells functionally synthesize a heterologous cytochrome P450 monooxygenase enabling cyclohexane hydroxylation. The biocatalyst‐specific reaction rate was found to be light‐dependent, reaching 26.3 ± 0.6 U gCDW‐1 at a light intensity of 150 µmolphotons m‐2 s‐1. In situ substrate supply via a two‐liquid phase system increased the initial specific activity to 39.2 ± 0.7 U gCDW‐1 and stabilized the biotransformation by preventing cell toxification. This resulted in a 10 times increased specific product yield of 4.5 gcyclohexanol gCDW‐1 as compared to the single aqueous phase system. Subsequently, the biotransformation was scaled from shake flask‐ to a 3 L stirred‐tank photo‐bioreactor setup. In situ O2 generation via photosynthetic water oxidation allowed non‐aerated process operation, thus circumventing substrate evaporation as the most critical factor limiting process performance and stability. This study for the first time exemplifies the technical applicability of cyanobacteria for aeration‐independent light‐driven oxyfunctionalization reactions involving highly toxic and volatile substrates.
ID 21866
Persistent UFZ Identifier http://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=21866
Hoschek, A., Toepel, J., Hochkeppel, A., Karande, R., Bühler, B., Schmid, A. (2019):
Light‐dependent and aeration‐independent gram‐scale hydroxylation of cyclohexane to cyclohexanol by CYP450 harboring Synechocystis sp. PCC 6803
Biotechnol. J. 14 (8), art. 1800724