Characterization of enzymatic D‐xylulose 5‐phosphate synthesis

Abstract

In this article we report on the characterization of the enzymatic synthesis of D-xylulose 5-phosphate using triosephosphate isomerase and transketolase. Two potential starting substrates are possible with this scheme. The data presented here allow a comparison of both routes for the synthesis, based on experimental information on reaction kinetics. Operational guidelines are proposed which should assist in the scale-up of such syntheses.

One‐Pot Cascade Reactions using Fructose‐6‐phosphate Aldolase: Efficient Synthesis of D‐Arabinose 5‐Phosphate, D‐Fructose 6‐Phosphate and Analogues

“…Selectively phosphorylated chiral sugars are of importance as central metabolites in the non-oxidative pentose phosphate pathway, in the Calvin cycle and of particular interest for the study of various metabolic diseases. [4] Given the vital roles for such compounds many methods have been developed for their synthesis. Enzymatic methods ensure mild reaction conditions and furthermore, tedious protection and deprotection of functional groups often resulting in low yields can be circumvented.…”

“…However efficient applications were illustrated in the preparative synthesis of glucose-6-phosphate [5a] and dihydroxyacetone phosphate (DHAP). [5b-c] Another alternative is the use of C-C bond formation catalysed by transketolase [4] alone, combined in cascade reaction with an aldolase [8] Numerous methods based on enzymatic reaction cascades were described for the preparation of DHAP, as well as chemical methods. [8c,9] Recently, two thermophilic aldolases (D-fructose-1,6-bisphosphate aldolase (FBA) and deoxyribose aldolase (DERA)) were combined in vivo (E. coli) for the synthesis of deoxyribose-5-phosphate in an artificial biosynthetic pathway.…”

Real‐time pH monitoring of industrially relevant enzymatic reactions in a microfluidic side‐entry reactor (μSER) shows potential for pH control

“…[19, 20]. The synthetic utility of transketolasecatalyzed reactions is based on the excellent selectivity and versatility of the two-carbon chain elongation of suitable aldehydes to more complex chiral compounds in combination with the irreversibility of this reaction by the use of hydroxypyruvate as carbon donor [21][22][23][24][25].…”

“…[19, 20]. The synthetic utility of transketolasecatalyzed reactions is based on the excellent selectivity and versatility of the two-carbon chain elongation of suitable aldehydes to more complex chiral compounds in combination with the irreversibility of this reaction by the use of hydroxypyruvate as carbon donor [21][22][23][24][25].In this contribution, we demonstrate real-time monitoring of pH in a microfluidic reactor as a further step towards their use as novel process development tools for biocatalysis. To achieve this, optical pH sensor layers with dual lifetime referencing [26] were integrated in a microfluidic side-entry reactor.…”

Production of biocommodities and bioelectricity by cell‐free synthetic enzymatic pathway biotransformations: Challenges and opportunities

“…For example, fructose is produced from glucose as catalyzed by glucose isomerase (Bhosale et al, 1996;Vasic-Racki, 2006), and beta-lactam antibiotics are semi-synthesized by amidases (Demain, 2004;Vasic-Racki, 2006). Relatively complicated chemical reactions can be mediated by multi-enzymes in one pot, for example, cellulose hydrolysis by the synergetic action of endoglucanase, cellobiohydrolase, and beta-glucosidase (Zhang and Lynd, 2004), synthesis of chiral alcohols with NAD(P)H regeneration (Hummel, 1999;Wichmann and Vasic-Racki, 2005), synthesis of carbohydrates (Meyer et al, 2007;Shaeri et al, 2008), and biosynthesis of polymers (Chi et al, 2008). The use of multiple enzymes in one pot has numerous benefits: fewer unit operations, smaller reactor volume, higher volumetric and space-time yields, shorter cycle times, and less waste generation.…”