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Title (Primary) Anaerobic C-H oxyfunctionalization: Coupling of nitrate reduction and quinoline hydroxylation in recombinant Pseudomonas putida
Author Ütkür, F.Ö.; Schmid, A.; Bühler, B.;
Journal Biotechnology Journal
Year 2019
Department SOMA;
Volume 14
Issue 8
Language englisch;
POF III (all) T15;
Supplements https://onlinelibrary.wiley.com/action/downloadSupplement?doi=10.1002%2Fbiot.201800615&file=biot201800615-sup-0001-%E2%94%9C%D0%ACtk%E2%94%9C%E2%95%9Dr_et_al_SI.docx
Keywords biocatalysis; industrial biotechnology; metabolic engineering; bacteria; biotransformation; cellular engineering; genetic engineering; redox biocatalysis; anaerobic respiration; hydroxylation
Abstract Whole‐cell biocatalysis for C‐H oxyfunctionalization depends on and often is limited by O2 mass transfer. In contrast to oxygenases, molybdenum hydroxylases use water instead of O2 as oxygen donor and thus have the potential to relieve O2 mass transfer limitations. Molybdenum hydroxylases may even allow anaerobic oxyfunctionalization when coupled to anaerobic respiration. To evaluate this option, the coupling of quinoline hydroxylation to denitrification was tested under anaerobic conditions employing Pseudomonas putida 86, capable of aerobic growth on quinoline. P. putida 86 reduced both nitrate and nitrite, but at low rates, which did not enable significant growth and quinoline hydroxylation. Introduction of the nitrate reductase from Pseudomonas aeruginosa enabled considerable specific quinoline hydroxylation activity (6.9 U gCDW‐1) under anaerobic conditions with nitrate as electron acceptor and 2‐hydroxyquinoline as the sole product (further metabolization depends on O2). Hydroxylation‐derived electrons were efficiently directed to nitrate, accounting for 38% of the respiratory activity. This study shows that molybdenum hydroxylase‐based whole‐cell biocatalysts enable completely anaerobic carbon oxyfunctionalization, when coupled to alternative respiration schemes such as nitrate respiration.
ID 21902
Persistent UFZ Identifier http://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=21902
Ütkür, F.Ö., Schmid, A., Bühler, B. (2019):
Anaerobic C-H oxyfunctionalization: Coupling of nitrate reduction and quinoline hydroxylation in recombinant Pseudomonas putida
Biotechnol. J. 14 (8), art. 1800615