Cascade of bioreactors in series for conversion of 3-phospho-d-glycerate into d-ribulose-1,5-bisphosphate: kinetic parameters of enzymes and operation variables

Coupled chemo(enzymatic) reactions in continuous flow

“…An illustrative example is the continuous conversion of D-3-phosphoglycerate ( 57 ) into D-ribulose 1,5-bisphosphate ( 58 ) in a series of eleven packed-bed reactors containing immobilized enzymes (Scheme 17) [41]. The aim of the process is to continuously regenerate D- 58 , which is used as an acceptor for the biocatalytic fixation of CO 2 resulting in the formation of two molecules of D- 57 .…”

“…E1: Phosphoglycerate kinase; E2: Glycerate phosphate dehydrogenase; E3: Triose phosphate isomerase; E4: Aldolase/FBP; E5: Phosphatase/FBPase; E6: Transketolase; E7: Aldolase/SuBP; E8: Phosphatase/SBPase; E9: Transketolase; E10: Epimerase; E11: Phosphoribulokinase [41]. …”

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“…After adjustment of pH using controllers and pH-stat the solution is fed to immobilized Rubisco reactors [18] where acceptor D-Ribulose-1,5-bisphosphate (RuBP) after CO 2 fixation is converted into 3-phosphoglycerate [16,17]. However, inasmuch as the recycling of acceptor RuBP is central to continuous CO 2 fixation, we have invented a novel scheme (Figure 1), which proceeds with no loss of CO 2 (unlike cellular biochemical systems) in 11 steps in a series of bioreactors [20]. This scheme is very different from generation of RuBP from D-glucose for start-up process [21] and employing 11 steps in different reactors requiring large volume and weight.…”

“…DNA sequencing of the plasmid identified the mutant. Recombinant wild-type and mutant (R38Q) 3-phosphoglycerate kinase (PGK) were purified to apparent homogeneity as described previously [20] have been shown in Figure 3A. The wild-type and mutant protein was incubated with 10 mM 3-phosphoglycerate barium salt (3PGA) in 50 mM Tris-Cl buffer, pH 7.5 containing 50 mM NaCl for overnight at room temperature.…”

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“…After adjustment of pH using controllers and pH-stat the solution is fed to immobilized Rubisco reactors [12] where acceptor D-Ribulose-1,5-bisphosphate (RuBP) after CO 2 fixation is converted into 3-phosphoglycerate [8,9]. We have invented a novel scheme which proceeds with no loss of CO 2 (unlike cellular biochemical systems) in 11 steps in a series of bioreactors [13]. …”