Enzyme‐catalyzed Aldol Additions

Fessner, Wolf‐Dieter

doi:10.1002/9783527619566.ch5

Cited by 39 publications

(9 citation statements)

References 286 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Directed aldol reaction is one of the most powerful carbon–carbon bond-forming procedures in synthetic organic chemistry and enables the concomitant creation of functionalized stereogenic centers and construction of chiral complex polyhydroxylated molecules [ 1 , 2 ]. Catalytic asymmetric addition of carbon nucleophiles (donor) to ketones (acceptor) is a fundamental approach to construct new tetrasubstituted stereogenic carbon centers.…”

Section: Introductionmentioning

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

View full text Add to dashboard Cite

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.

show abstract

Section: Introductionmentioning

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

View full text Add to dashboard Cite

show abstract

“…The most prominent examples are the transketolases, oxynitrilases and aldolases [1,2]. For biocatalytic applications a range of aldolases have already been examined and found useful for the stereo-controlled formation of carbon-carbon bonds [3][4][5][6]. In addition, some aldolases affect the resolution of racemates, by specifically cleaving the carbon-carbon bond of only one enantiomer [7,8].…”

Section: Introductionmentioning

confidence: 99%

Directed evolution of an industrial biocatalyst: 2‐deoxy‐D‐ribose 5‐phosphate aldolase

Jennewein

Schürmann

Wolberg

et al. 2006

Biotechnology Journal

161

131

View full text Add to dashboard Cite

Aldolases are emerging as powerful and cost efficient tools for the industrial synthesis of chiral molecules. They catalyze enantioselective carbon-carbon bond formations, generating up to two chiral centers under mild reaction conditions. Despite their versatility, narrow substrate ranges and enzyme inactivation under synthesis conditions represented major obstacles for large-scale applications of aldolases. In this study we applied directed evolution to optimize Escherichia coli 2-deoxy-D-ribose 5-phosphate aldolase (DERA) as biocatalyst for the industrial synthesis of (3R,5S)-6-chloro-2,4,6-trideoxyhexapyranoside. This versatile chiral precursor for vastatin drugs like Lipitor (atorvastatin) is synthesized by DERA in a tandem-aldol reaction from chloroacetaldehyde and two acetaldehyde equivalents. However, E. coli DERA shows low affinity to chloroacetaldehyde and is rapidly inactivated at aldehyde concentrations useful for biocatalysis. Using high-throughput screenings for chloroacetaldehyde resistance and for higher productivity, several improved variants have been identified. By combination of the most beneficial mutations we obtained a tenfold improved variant compared to wild-type DERA with regard to (3R,5S)-6-chloro-2,4,6-trideoxyhexapyranoside synthesis, under industrially relevant conditions.

show abstract

“…Biocatalysis by means of aldolases offers a unique stereoselective and green tool to perform this transformation [2,3].…”

Section: Introductionmentioning

confidence: 99%

In situ aldehyde generation for aldol addition reactions catalyzed by d-fructose-6-phosphate aldolase

Mifsud

Szekrényi

Joglar

et al. 2012

Journal of Molecular Catalysis B: Enzymatic

View full text Add to dashboard Cite

Enzyme‐catalyzed Aldol Additions

Cited by 39 publications

References 286 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

Directed evolution of an industrial biocatalyst: 2‐deoxy‐D‐ribose 5‐phosphate aldolase

In situ aldehyde generation for aldol addition reactions catalyzed by d-fructose-6-phosphate aldolase

Contact Info

Product

Resources

About