Development of in-situ product removal strategies in biocatalysis applying scaled-down unit operations

Publication: Research - peer-reviewJournal article – Annual report year: 2016

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An experimental platform based on scaled-down unit operations combined in a plug-and-play manner enables easy and highly flexible testing of advanced biocatalytic process options such as in-situ product removal (ISPR) process strategies. In such a platform it is possible to compartmentalize different process steps while operating it as a combined system, giving the possibility to test and characterize the performance of novel process concepts and biocatalysts with minimal influence of inhibitory products. Here the capabilities of performing process development by applying scaled-down unit operations are highlighted through a case study investigating the asymmetric synthesis of 1-methyl-3-phenylpropylamine (MPPA) using ω-transaminase, an enzyme in the sub-family of amino transferases (ATAs). An on-line HPLC system was applied to avoid manual sample handling and to semi-automatically characterize ω-transaminases in a scaled-down packed-bed reactor (PBR) module, showing MPPA as a strong inhibitor. To overcome the inhibition, a two-step liquid-liquid extraction (LLE) ISPR concept was tested using scaled-down unit operations combined in a plug-and-play manner. Through the tested ISPR concept, it was possible to continuously feed the main substrate benzylacetone (BA) and extract the main product MPPA throughout the reaction, thereby overcoming the challenges of low substrate solubility and product inhibition. The tested ISPR concept achieved a product concentration of 26.5 gMPPA·L–1, a purity up to 70% gMPPA·L–1 and a recovery in the range of 80% mol·mol–1 of MPPA in 20 hours, with the possibility to increase the concentration, purity and recovery further. This article is protected by copyright. All rights reserved
Original languageEnglish
JournalBiotechnology and Bioengineering
Volume114
Issue number3
Pages (from-to)600–609
ISSN0006-3592
DOIs
StatePublished - 2017
CitationsWeb of Science® Times Cited: 0

    Keywords

  • In-situ product removal (ISPR), Substrate supply, Scale-down unit operations, Biocatalysis, Process intensification, Chiral amines, Amino transferase (ATAs)
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