TY  - JOUR
AU  - Oeggl, Reinhard
AU  - Neumann, Timo
AU  - Gätgens, Jochem
AU  - Romano, Diego
AU  - Noack, Stephan
AU  - Rother, Dörte
TI  - Citrate as Cost-Efficient NADPH Regenerating Agent
JO  - Frontiers in Bioengineering and Biotechnology
VL  - 6
SN  - 2296-4185
CY  - Lausanne
PB  - Frontiers Media
M1  - FZJ-2018-07409
SP  - 196
PY  - 2018
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:30631764
UR  - <Go to ISI:>//WOS:000454341200001
DO  - DOI:10.3389/fbioe.2018.00196
UR  - https://juser.fz-juelich.de/record/858538
ER  -