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000858538 1001_ $$0P:(DE-Juel1)168384$$aOeggl, Reinhard$$b0
000858538 245__ $$aCitrate as Cost-Efficient NADPH Regenerating Agent
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000858538 520__ $$aThe economically efficient utilization of NADPH or NADH-dependent enzymes requires the regeneration of consumed reduction equivalents. This cofactor regeneration is classically done via an additional substrate, and if necessary enzyme. We now demonstrate an easy-to-apply cofactor regeneration approach, which can especially be used in screening applications. Simply by applying citrate to a buffer or directly using citrate/-phosphate buffer NADPH can be regenerated by native enzymes of the TCA cycle, practically present in all aerobic living organisms. Apart from viable-culturable cells, this regeneration approach can also be applied with lyophilized cells and even crude cell extracts. This is exemplarily shown for the synthesis of 1 phenylethanol from acetophenone with several oxidoreductases. The mechanism of NADPH regeneration by TCA cycle enzymes was further investigated by a transient isotopic labeling experiment feeding [1,5-13C]citrate. This revealed that the regeneration mechanism can further be optimized by genetic modification of two competing internal citrate metabolization pathways, the glyoxylate shunt and the glutamate dehydrogenase.
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000858538 7001_ $$0P:(DE-Juel1)169334$$aNeumann, Timo$$b1
000858538 7001_ $$0P:(DE-Juel1)129023$$aGätgens, Jochem$$b2
000858538 7001_ $$0P:(DE-HGF)0$$aRomano, Diego$$b3
000858538 7001_ $$0P:(DE-Juel1)129050$$aNoack, Stephan$$b4
000858538 7001_ $$0P:(DE-Juel1)144643$$aRother, Dörte$$b5$$eCorresponding author
000858538 773__ $$0PERI:(DE-600)2719493-0$$a10.3389/fbioe.2018.00196$$gVol. 6, p. 196$$p196$$tFrontiers in Bioengineering and Biotechnology$$v6$$x2296-4185$$y2018
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