Single-pot enzymatic reaction sequence for the synthesis of D-glyceraldehyde-3-phosphate

Getachew Shibabaw Molla, P. J. Venkatesh, A. Liese

Research output: Contribution to journalConference abstract in journalResearchpeer-review


A single-pot enzymatic reaction sequence has been designed for the synthesis of a key intermediate metabolite called D-glyceraldehyde-3-phosphate(D-GAP). The reaction sequence consistsof three enzymes and usesD-F16BP as a starting material (Fig.).The rabbit muscle aldolase (RAMA)-catalyzed reaction step is in favor of D-F16BP formation with an equilibrium constant of 10–4M [1]. Therefore, the single-pot reaction sequence has been designed to shift the equilibrium and regenerate NADH. The capability of the reaction sequence was demonstrated by the enhanced equilibrium conversion from 1.4% to 96%. The reaction system was optimized in detail with regard to activity and stability of the enzymes and stability of the cofactors (NADH and NAD+) and D-GAP. Reaction kinetics models for each of the enzymes were formulated. The effect of co-substances on the activity ofthe enzymes was evaluated. The results elucidated that D-F16BP and sn-G3P do not influence the activity of FDH and HCOO does not affect the activity of RAMA, whereas D-F16BP and HCOOsuppress the activity of sn-G3PDH. From the kinetics perspective and due to the stability of the enzymes, a continuous operation based on an enzyme membrane reactor was preferred
Original languageEnglish
Issue number9
Pages (from-to)1401–1426
Publication statusPublished - 2014
Externally publishedYes
EventProcessNet-Jahrestagung und 31. DECHEMA-Jahrestagung der Biotechnologen 2014 - Aachen, Germany
Duration: 30 Sept 20142 Oct 2014


ConferenceProcessNet-Jahrestagung und 31. DECHEMA-Jahrestagung der Biotechnologen 2014


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