Enzyme-catalysed carbon-carbon bond formation: Large-scale production of Escherichia coli transketolase

Hobbs, G. R.; Mitra, Robin; Chauhan, Rashmi P.; Woodley, John M.; Lilly, M. D.

doi:10.1016/0168-1656(95)00165-4

Cited by 39 publications

(17 citation statements)

References 14 publications

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“…The three-dimensional structure of the latter enzyme [93,111] has been published (PDB accession code 1TRK) and crystallographic studies on the E. coli enzyme have commenced [94]. Homologous expression systems are established for the S. cerevisiae [155] and the E. coli [60,153] enzyme. For human TK two laboratories have reported [140] or made use of a bacterial heterologous expression system [149,171].…”

Section: Sources Of Tkmentioning

confidence: 99%

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Properties and functions of the thiamin diphosphate dependent enzyme transketolase

Schenk

Duggleby

Nixon

1998

The International Journal of Biochemistry & Cell Biology

223

164

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This review highlights recent research on the properties and functions of the enzyme transketolase, which requires thiamin diphosphate and a divalent metal ion for its activity. The transketolase-catalysed reaction is part of the pentose phosphate pathway, where transketolase appears to control the non-oxidative branch of this pathway, although the overall¯ux of labelled substrates remains controversial. Yeast transketolase is one of several thiamin diphosphate dependent enzymes whose three-dimensional structures have been determined. Together with mutational analysis these structural data have led to detailed understanding of thiamin diphosphate catalysed reactions. In the homodimer transketolase the two catalytic sites, where dihydroxyethyl groups are transferred from ketose donors to aldose acceptors, are formed at the interface between the two subunits, where the thiazole and pyrimidine rings of thiamin diphosphate are bound. Transketolase is ubiquitous and more than 30 full-length sequences are known. The encoded protein sequences contain two motifs of high homology; one common to all thiamin diphosphate-dependent enzymes and the other a unique transketolase motif. All characterised transketolases have similar kinetic and physical properties, but the mammalian enzymes are more selective in substrate utilisation than the nonmammalian representatives. Since products of the transketolase-catalysed reaction serve as precursors for a number of synthetic compounds this enzyme has been exploited for industrial applications. Putative mutant forms of transketolase, once believed to predispose to disease, have not stood up to scrutiny. However, a modi®cation of transketolase is a marker for Alzheimer's disease, and transketolase activity in erythrocytes is a measure of thiamin nutrition. The cornea contains a particularly high transketolase concentration, consistent with the proposal that pentose phosphate pathway activity has a role in the removal of light-generated radicals. #

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Section: Sources Of Tkmentioning

confidence: 99%

“…TK is also an important biocatalyst in stereospeci®c carbon±carbon bond synthesis [60]. TK accepts a large number of aldehydes as substrates [59,67,106], in particular those containing an a-hydroxy group in 2R con®guration; the products are ketoses with a 3 S,4R con®gur-ation (threo con®guration) [27,31].…”

Section: Industrial Uses Of Tkmentioning

confidence: 99%

Properties and functions of the thiamin diphosphate dependent enzyme transketolase

Schenk

Duggleby

Nixon

1998

The International Journal of Biochemistry & Cell Biology

223

164

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“…Use of b-hydroxypyruvate (b-HPA) as the donor substrate, renders the reaction irreversible (de la Haba et al 1955), and improves its potential as a catalyst for large-scale biotransformations (French et al 1995;Hobbs et al 1996;Brocklebank et al 1999;Shaeri et al 2006). Several TK activity assays could potentially be implemented in microplates.…”

Section: Introductionmentioning

confidence: 99%

Optimisation and evaluation of a generic microplate-based HPLC screen for transketolase activity

et al. 2007

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A microplate-based HPLC assay for transketolase is described for rapidly determining substrate and product concentration suitable for optimisation of biocatalytic process conditions and screening directed evolution libraries. Transketolase catalyses the enantioselective carbon-carbon bond formation of chiral keto-diol products. The assay was used to determine dissociation constants for the two cofactors required by transketolase with 5-11% error. The preparation of samples by microplate-based fermentation, cell lysis, addition of cofactor, addition of substrates was also evaluated and optimised for increased transketolase activity. The whole process enables 3-fold improved enzyme variants to be identified from a single measurement.

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“…1). Transketolase is now also available (Hobbs et al, 1996) so that it becomes feasible to consider such a scheme (Seoane, 2000). Interestingly, the starting point can be either DHAP itself or glyceraldehyde 3-phosphate (GA3P).…”

Section: Introductionmentioning

confidence: 99%

Characterization of enzymatic D‐xylulose 5‐phosphate synthesis

Shaeri

Wright²,

Rathbone³

et al. 2008

Biotech & Bioengineering

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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.

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Enzyme-catalysed carbon-carbon bond formation: Large-scale production of Escherichia coli transketolase

Cited by 39 publications

References 14 publications

Properties and functions of the thiamin diphosphate dependent enzyme transketolase

Properties and functions of the thiamin diphosphate dependent enzyme transketolase

Optimisation and evaluation of a generic microplate-based HPLC screen for transketolase activity

Characterization of enzymatic D‐xylulose 5‐phosphate synthesis

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