Cell-free biocatalytic syntheses of L-pipecolic acid: a dual strategy approach and process intensification in flow

Roura Padrosa, David; Benítez-Mateos, Ana I.; Calvey, Liam; Paradisi, Francesca (2020). Cell-free biocatalytic syntheses of L-pipecolic acid: a dual strategy approach and process intensification in flow. Green chemistry, 22(16), pp. 5310-5316. Royal Society of Chemistry 10.1039/D0GC01817A

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As an alternative to the traditional chemical synthesis or in-vivo production of L-Pipecolic acid, we have developed two ex-vivo strategies using purified and immobilised enzymes for the production of this key building block. Firstly, a transaminase capable of lysine ε-deamination was coupled with a novel pyrroline-5-carboxylate reductase, yielding 60% conversion at the 50 mM scale with free enzymes and in-situ recycling of the cofactor. A second, simpler, redox neutral system was then constructed by combining the pyrroline-5-carboxylate reductase with a lysine-6-dehydrogenase. This bienzymatic system, with catalytic amount of free cofactor yielded >99% of pipecolic acid in batch and, following co-immobilisation of both enzymes, it was applied as a packed-bed reactor in continuous flow achieving again a molar conversion of >99% with 30 min residence time, and a space-time yield up to 2.5 g/L/h. The sustainability of the system was further improved by a catch-and-release strategy to purify the product, and recovery and recycling of the cofactor.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Roura Padrosa, David; Benitez Mateos, Ana Isabel and Paradisi, Francesca

Subjects:

500 Science > 540 Chemistry

ISSN:

1463-9262

Publisher:

Royal Society of Chemistry

Language:

English

Submitter:

Francesca Paradisi

Date Deposited:

23 Jul 2020 10:20

Last Modified:

19 Sep 2020 17:35

Publisher DOI:

10.1039/D0GC01817A

BORIS DOI:

10.7892/boris.145319

URI:

https://boris.unibe.ch/id/eprint/145319

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