DHAP-dependent aldolases from (hyper)thermophiles: biochemistry and applications

Falcicchio, Pierpaolo; Loo, Suzanne Wolterink-van; Franssen, M.C.R.; Oost, John van der

doi:10.1007/s00792-013-0593-x

Cited by 22 publications

(9 citation statements)

References 112 publications

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“…DHAP-dependent aldolases have been widely investigated for the synthesis of several new deoxy or phosphorylated sugars and iminocyclitols [ 22 – 24 ]. Naturally, this class of enzyme utilizes DHAP as the donor substrate and accepts a broad range of acceptor aldehydes.…”

Section: Resultsmentioning

confidence: 99%

Biosynthesis of dendroketose from different carbon sources using in vitro and in vivo metabolic engineering strategies

Yang

Zhu

et al. 2018

Biotechnol Biofuels

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BackgroundAsymmetric aldol-type C–C bond formation with ketones used as electrophilic receptor remains a challenging reaction for aldolases as biocatalysts. To date, only one kind of dihydroxyacetone phosphate (DHAP)-dependent aldolases has been discovered and applied to synthesize branched-chain sugars directly using DHAP and dihydroxyacetone (DHA) as substrate. However, the unstable and high-cost properties of DHAP limit large-scale application. Therefore, biosynthesis of branched-chain sugar from low-cost and abundant carbon sources is essential.ResultsThe detailed catalytic property of l-rhamnulose-1-phosphate aldolase (RhaD) and l-fuculose-1-phosphate aldolase (FucA) from Escherichia coli in catalyzing the aldol reactions with DHA as electrophilic receptors was characterized. Furthermore, we calculated the Bürgi–Dunitz trajectory using molecular dynamics simulations, thereby revealing the original sources of the catalytic efficiency of RhaD and FucA. A multi-enzyme reaction system composed of formolase, DHA kinase, RhaD, fructose-1-phosphatase, and polyphosphate kinase was constructed to in vitro produce dendroketose, a branched-chain sugar, from one-carbon formaldehyde. The conversion rate reached 86% through employing a one-pot, two-stage reaction process. Moreover, we constructed two artificial pathways in Corynebacterium glutamicum to obtain this product in vivo starting from glucose or glycerol. Fermentation with glycerol as feedstock produced 6.4 g/L dendroketose with a yield of 0.45 mol/mol glycerol, representing 90% of the maximum theoretical value. Additionally, the dendroketose production reached 36.3 g/L with a yield of 0.46 mol/mol glucose when glucose served as the sole carbon resource.ConclusionsThe detailed enzyme kinetics data of the two DHAP-dependent aldolases with DHA as electrophilic receptors were presented in this study. In addition, insights into this catalytic property were given via in silico simulations. Moreover, the cost-effective synthesis of dendroketose starting from one-, three-, and six-carbon resources was achieved through in vivo and in vitro metabolic engineering strategies. This rare branched-chain ketohexose may serve as precursor to prepare 4-hydroxymethylfurfural and branched-chain alkanes using chemical method.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1293-7) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Biosynthesis of dendroketose from different carbon sources using in vitro and in vivo metabolic engineering strategies

Yang

Zhu

et al. 2018

Biotechnol Biofuels

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“…Despite these advantages, DHAP-dependent aldolases from hyperthermophilic organisms have been barely used in synthesis (Falcicchio et al 2014). Recent example are those described by Wever group, in which rhamnulose-1-phosphate aldolase (Rha-1PA) from Thermotoga maritima -a hyperthermophilic eubacterium with an optimal growth temperature of 80 °Cwas used soluble in a one-pot four-enzyme catalytic cascade for the simple and inexpensive synthesis of a variety of non-natural heterocycles or immobilized on epoxy beads in a continuous flow reactor for the synthesis of various aldol products (Babich et al 2011;Babich et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima

Oroz‐Guinea

Sánchez-Moreno

Mena³

et al. 2014

Appl Microbiol Biotechnol

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The TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PA Tm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co(2+) as a divalent metal ion cofactor, instead of Zn(2+) as its E. coli homologue, and exhibited a maximum of activity at 95 °C. Furthermore, the enzyme displayed a high stability against extreme reaction conditions, retaining 90 % of its activity in the presence of 40 % of acetonitrile and showing a half-life greater than 3 h at 115 °C. The kinetic parameters at room temperature (R/T) were also studied; the K M was calculated to be 3.6 ± 0.33 mM, while k cat/K M was found to be 0.7 × 10(3) s(-1) M(-1). Given these characteristics, Rha-1PA Tm is an attractive enzyme for use as a biocatalyst for industrial applications, offering intriguing possibilities for practical biocatalysis.

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“…Thus there is a real need to find new aldolases able to display broader donor specificities, in particular, for nonphosphorylated substrates to increase the chemist’s toolbox of versatile biocatalysts 6c. g The discovery of fructose‐6‐phosphate aldolases (FSAA coli and FSAB coli ) in Escherichia coli has opened up exciting new perspectives 7. Indeed, FSAs coli possesses the unique capacity to react efficiently with at least four different non‐phosphorylated donor substrates (dihydroxyacetone, DHA; hydroxyacetone, HA; hydroxybutanone and glycolaldehyde) and various acceptors 7a.…”

Section: Introductionmentioning

confidence: 99%

“…8 Given the extraordinary potential of FSAs, using wild‐type or variants, many syntheses were described to prepare especially antidiabetic (glycosidase inhibitors), anti‐infective compounds or phosphorylated sugars as metabolites 6c. 9 Since their discovery in 2001, no new natural efficient DHA aldolases have been identified6g from other microorganisms unlike most other aldolases. However, E. coli transaldolase B (TalB coli ) and DHAP‐dependent rhamnulose aldolase display a slight aldol promiscuous activity towards DHA that has been improved by mutagenesis 10…”

Section: Introductionmentioning

confidence: 99%

Genome Mining for Innovative Biocatalysts: New Dihydroxyacetone Aldolases for the Chemist’s Toolbox

et al. 2015

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Stereoselective carboligating enzymes were discovered by a genome mining approach to extend the biocatalysis toolbox. Seven hundred enzymes were selected by sequence comparison from diverse prokaryotic species as representatives of the aldolase (FSA) family diversity. The aldol reaction tested involved dihydroxyacetone (DHA) and glyceraldehyde‐3‐phosphate. The hexose‐6‐phosphate formation was monitored by mass spectrometry. Eighteen enzymes annotated either as transaldolases or aldolases were found to exhibit a DHA aldolase activity. Remarkably, six of them proven as aldolases, and not transaldolases, shared very limited similarities with those currently described. Multiple sequence alignment performed on all enzymes revealed a Tyr in the new DHA aldolases as found in FSAcoli instead of a Phe usually found in transaldolases. Four of these DHA aldolases were biochemically characterised in comparison with FSAcoli. In particular, an aldolase from Listeria monocytogenes exhibited interesting catalytic properties.

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DHAP-dependent aldolases from (hyper)thermophiles: biochemistry and applications

Cited by 22 publications

References 112 publications

Biosynthesis of dendroketose from different carbon sources using in vitro and in vivo metabolic engineering strategies

Biosynthesis of dendroketose from different carbon sources using in vitro and in vivo metabolic engineering strategies

Hyperthermophilic aldolases as biocatalyst for C–C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima

Genome Mining for Innovative Biocatalysts: New Dihydroxyacetone Aldolases for the Chemist’s Toolbox

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