Enzymatic synthesis of rare sugars with l-rhamnulose-1-phosphate aldolase from Thermotoga maritima MSB8

Li, Zijie; Wu, Xuehong; Cai, Li; Duan, Shenglin; Liu, Jia; Yuan, Peng; Nakanishi, Hideki; Gao, Xiao‐Dong

doi:10.1016/j.bmcl.2015.07.027

Cited by 11 publications

(5 citation statements)

References 15 publications

(18 reference statements)

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“…Apart from the “Izumoring strategy”, aldol additions catalyzed by aldolases afford a promising alternative for rare sugar synthesis through joining two smaller fragments. − Among all aldolases, the dihydroxyacetone phosphate (DHAP)-dependent aldolases are very popularly used for building carbon–carbon bonds in synthesizing carbohydrates and their derivatives. − In our previous study, we adopted a one-pot four-enzyme system based on different DHAP-dependent aldolases and successfully used it to produce various rare ketoses. − In this system (Scheme ), we had to supply both fragments for the aldol reaction: glycerol 3-phosphate was used to produce DHAP through oxidation by glycerol phosphate oxidase (GPO), whereas the acceptor aldehyde had to be added externally. The by-product hydrogen peroxide from the oxidation step must be degraded into oxygen and water by catalase.…”

Section: Introductionmentioning

confidence: 99%

One-Pot Multienzyme Synthesis of Rare Ketoses from Glycerol

Cai

et al. 2020

J. Agric. Food Chem.

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A facile approach is introduced here for the synthesis of rare ketoses from glycerol and d-/l-glyceraldehyde (d-/l-GA). The reactions were carried out in a one-pot multienzyme fashion in which the only carbon source is glycerol. In the enzymatic cascade, glycerol is phosphorylated and then oxidized at C2 to afford dihydroxyacetone phosphate (DHAP), the key donor for enzymatic aldol reaction. Meanwhile, the primary alcohol of glycerol is also oxidized to give the acceptor molecule GA in situ (d- or l-isomer could be formed stereospecifically with either alditol oxidase or horse liver alcohol dehydrogenase). Different DHAP-dependent aldolases were used to generate the aldol adducts (rare ketohexose phosphates) with various stereoconfigurations and diastereomeric ratios. It is worth noting that the enzyme that catalyzes the phosphorylation reaction in the first step could also help recycle the phosphate in the last step to provide free rare sugar molecules. This study provides a useful method for rare ketose synthesis on a 100 mg to g scale, starting from relatively inexpensive materials which solved the problem of supplying both glycerol 3-phosphate and GA in our previous work. It also demonstrates an example of green synthesis due to highly efficient carbon usage and recycling of cofactors.

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Section: Introductionmentioning

confidence: 99%

One-Pot Multienzyme Synthesis of Rare Ketoses from Glycerol

Cai

et al. 2020

J. Agric. Food Chem.

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“…It should be noted that the d ‐allulose and d ‐sorbose production by the control strain should be a result of the basal expression of RhaD , FucA , and YqaB from the genomic DNA (Figure 2B,C). Excluding the activities of RhaD, FucA, and YqaB from the genome, we concluded that RhaD‐YqaB produced both d ‐allulose and d ‐sorbose, [ 28,46 ] while FucA‐YqaB produced only d ‐allulose. [ 29 ] These results indicated that both RhaD and FucA worked as aldolases, and YqaB worked as a phosphatase.…”

Section: Resultsmentioning

confidence: 99%

Engineering an efficient whole‐cell catalyst for d‐allulose production from glycerol

Zhang

Zhao

et al. 2023

Biotechnology Journal

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d‐Allulose has many health‐benefiting properties and therefore sustainably applied in food, pharmaceutical, and nutrition industries. The aldol reaction‐based route is a very promising alternative to Izumoring strategy in d‐allulose production. Remarkable studies reported in the past cannot get rid of by‐product formation and costly purified enzyme usage. In the present study, we explored the glycerol assimilation by modularly assembling the d‐allulose synthetic cascade in Escherichia coli envelop. We achieved an efficient whole‐cell catalyst that produces only d‐allulose from cheap glycerol feedstock, eliminating the involvement of purified enzymes. Detailed process optimization improved the d‐allulose titer by 1500.00%. Finally, the production was validated in 3‐L scale using a 5‐L fermenter, and 5.67 g L−1 d‐allulose was produced with a molar yield of 31.43%.

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“…We further investigated whether this unexpected reaction is common within the RhuA family.W eu sed as equencedriven approach [13] to build as mall library representative of the functional diversity of the RhuA family,u sing experimentally confirmed RhuA [14,15] and C2JUR4 from Lactobacillus rhamnosus as references.W es elected at otal of 21 proteins,all annotated as RhuA, with ahigh degree of amino acid sequence diversity (Table S1). Their corresponding genes were cloned (Table S2) and the proteins overexpressed.…”

Section: Methodsmentioning

confidence: 99%

Synthesis of Branched‐Chain Sugars with a DHAP‐Dependent Aldolase: Ketones are Electrophile Substrates of Rhamnulose‐1‐phosphate Aldolases

et al. 2018

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Dihydroxyacetone phosphate (DHAP)-dependent rhamnulose aldolases display an unprecedented versatility for ketones as electrophile substrates. We selected and characterized a rhamnulose aldolase from Bacteroides thetaiotaomicron (RhuABthet) to provide a proof of concept. DHAP was added as a nucleophile to several α-hydroxylated ketones used as electrophiles. This aldol addition was stereoselective and produced branched-chain monosaccharide adducts with a tertiary alcohol moiety. Several aldols were readily obtained in good to excellent yields (from 76 to 95 %). These results contradict the general view that aldehydes are the only electrophile substrates for DHAP-dependent aldolases and provide a new C-C bond-forming enzyme for stereoselective synthesis of tertiary alcohols.

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Enzymatic synthesis of rare sugars with l-rhamnulose-1-phosphate aldolase from Thermotoga maritima MSB8

Cited by 11 publications

References 15 publications

One-Pot Multienzyme Synthesis of Rare Ketoses from Glycerol

One-Pot Multienzyme Synthesis of Rare Ketoses from Glycerol

Engineering an efficient whole‐cell catalyst for d‐allulose production from glycerol

Synthesis of Branched‐Chain Sugars with a DHAP‐Dependent Aldolase: Ketones are Electrophile Substrates of Rhamnulose‐1‐phosphate Aldolases

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