Issue 16, 2020

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

Abstract

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−1 h−1. 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.

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

Supplementary files

Article information

Article type
Paper
Submitted
28 5月 2020
Accepted
21 7月 2020
First published
21 7月 2020
This article is Open Access
Creative Commons BY license

Green Chem., 2020,22, 5310-5316

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

D. Roura Padrosa, A. I. Benítez-Mateos, L. Calvey and F. Paradisi, Green Chem., 2020, 22, 5310 DOI: 10.1039/D0GC01817A

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