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Hollmann, F.

Publications and source records attributed to Hollmann, F..

2 recordsLinked to original sources

The atmospheric pressure capillary plasma jet is well-suited to supply H2O2 for plasma-driven biocatalysis

Plasma-generated H2O2 can be used to fuel biocatalytic reactions that require H2O2 as co-substrate such as the conversion of ethylbenzene to (R)-1-phenylethanol ((R)-1-PhOl) catalyzed by unspecific peroxygenase from Agrocybe aegerita (rAaeUPO). Immobilization was recently shown to protect biocatalysts from inactivation by highly reactive plasma-produced species, however, H2O2 supply by the employed plasma sources ({micro}APPJ and DBD) was limiting for rAaeUPO performance. In this study we evaluated a recently introduced capillary plasma jet for suitability to supply H2O2 in situ. H2O2 production was modulated by varying the water concentration in the feed gas, providing a greater operating window for applications in plasma-driven biocatalysis. In a static system after 80 min of biocatalysis, a turnover number of 44,199 mol(R)-1-PhOl mol-1rAaeUPO was achieved without significant enzyme inactivation. By exchanging the reaction solution every 5 min, a total product yield of 122 {micro}mol (R)-1-PhOl was achieved in 700 min run time, resulting in a total turnover number of 174,209 mol(R)-1-PhOl mol-1rAaeUPO. We conclude that the capillary plasma jet, due to its flexibility regarding feed gas, admixtures, and power input, is well-suited for in situ H2O2 generation for plasma-driven biocatalysis tailoring to enzymes with high H2O2 turnover.

biochemistry↗

A versatile in situ cofactor enhancing system for meeting cellular demands for engineered metabolic pathways

Cofactor imbalance obstructs the productivities of metabolically engineered cells. Herein, we employed a minimally perturbing system, xylose reductase and lactose (XR/lactose), to increase levels of a pool of sugar-phosphates which are connected to the biosynthesis of NAD(P)H, FAD, FMN and ATP in Escherichia coli. The XR/lactose system could increase the amounts of the precursors of these cofactors and was tested with three different metabolically engineered cell systems (fatty alcohol biosynthesis, bioluminescence light generation and alkane biosynthesis) with different cofactor demands. Productivities of these cells were increased 2-4-fold by the XR/lactose system. Untargeted metabolomic analysis revealed different metabolite patterns among these cells; demonstrating that only metabolites involved in relevant cofactor biosynthesis were altered. The results were also confirmed by transcriptomic analysis. Another sugar reducing system (glucose dehydrogenase, GDH) could also be used to increase fatty alcohol production but resulted in less yield enhancement than XR. This work demonstrates that the approach of increasing cellular sugar phosphates can be a generic tool to increase in vivo cofactor generation upon cellular demand for synthetic biology. TeaserUse of sugar and sugar reductase to increase sugar phosphates for enhancing in situ synthesis of cofactors upon cellular demand for synthetic biology.

synthetic biology↗