Journal Article

FZJ-2018-07409

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Citrate as Cost-Efficient NADPH Regenerating Agent

Oeggl, R.FZJ* ; Neumann, T. ; Gätgens, J.FZJ* ; Romano, D. ; Noack, S.FZJ* ; Rother, D. (Corresponding author)FZJ*

2018
Frontiers Media Lausanne

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Please use a persistent id in citations: http://hdl.handle.net/2128/21137  doi:10.3389/fbioe.2018.00196

Abstract: The economically efficient utilization of NADPH or NADH-dependent enzymes requires the regeneration of consumed reduction equivalents. This cofactor regeneration is classically done via an additional substrate, and if necessary enzyme. We now demonstrate an easy-to-apply cofactor regeneration approach, which can especially be used in screening applications. Simply by applying citrate to a buffer or directly using citrate/-phosphate buffer NADPH can be regenerated by native enzymes of the TCA cycle, practically present in all aerobic living organisms. Apart from viable-culturable cells, this regeneration approach can also be applied with lyophilized cells and even crude cell extracts. This is exemplarily shown for the synthesis of 1 phenylethanol from acetophenone with several oxidoreductases. The mechanism of NADPH regeneration by TCA cycle enzymes was further investigated by a transient isotopic labeling experiment feeding [1,5-13C]citrate. This revealed that the regeneration mechanism can further be optimized by genetic modification of two competing internal citrate metabolization pathways, the glyoxylate shunt and the glutamate dehydrogenase.

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Contributing Institute(s):

1. Biotechnologie (IBG-1)

Research Program(s):

1. 581 - Biotechnology (POF3-581) (POF3-581)

Appears in the scientific report 2018

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Database coverage:
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