Catalyses a step in the 3-hydroxypropanoate/4-hydroxybutanoate cycle, an autotrophic CO2 fixation pathway found in some thermoacidophilic archaea . The enzyme from Metallosphaera sedula acts nearly equally as well on (S)-3-hydroxybutanoyl-CoA but not (R)-3-hydroxybutanoyl-CoA .
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SYSTEMATIC NAME
IUBMB Comments
3-hydroxypropionyl-CoA hydro-lyase
Catalyses a step in the 3-hydroxypropanoate/4-hydroxybutanoate cycle, an autotrophic CO2 fixation pathway found in some thermoacidophilic archaea [1]. The enzyme from Metallosphaera sedula acts nearly equally as well on (S)-3-hydroxybutanoyl-CoA but not (R)-3-hydroxybutanoyl-CoA [2].
substrate specificity of enzyme Ms3HPCD, modelling of 3-hydroxypropanoyl- and (S)-3-hydroxybutyryl-moiety binding mode of enzyme Ms3HPCD. The residues involved in the formation of the 3-hydroxypropanoate binding pocket are identified. Ms3HPCD cannot convert (R)-stereoisomer of 3-hydroxybutyryl-CoA. When (R)-3-hydroxybutyryl-CoA is used as a substrate, the positions of the 3-hydroxyl-group and the C4-moiety are reversed each other, resulting in improper positioning of the (R)-3-hydroxybutyryl-moiety in the pocket. Ms3HPCD has a tightly formed alpha3 helix near the active site, and bulky aromatic residues are located at the enoyl-group binding site, resulting in the enzyme having an optimal substrate binding site for accepting short-chain 3-hydroxyacyl-CoA as a substrate
substrate specificity of enzyme Ms3HPCD, modelling of 3-hydroxypropanoyl- and (S)-3-hydroxybutyryl-moiety binding mode of enzyme Ms3HPCD. The residues involved in the formation of the 3-hydroxypropanoate binding pocket are identified. Ms3HPCD cannot convert (R)-stereoisomer of 3-hydroxybutyryl-CoA. When (R)-3-hydroxybutyryl-CoA is used as a substrate, the positions of the 3-hydroxyl-group and the C4-moiety are reversed each other, resulting in improper positioning of the (R)-3-hydroxybutyryl-moiety in the pocket. Ms3HPCD has a tightly formed alpha3 helix near the active site, and bulky aromatic residues are located at the enoyl-group binding site, resulting in the enzyme having an optimal substrate binding site for accepting short-chain 3-hydroxyacyl-CoA as a substrate
substrate specificity of enzyme Ms3HPCD, modelling of 3-hydroxypropanoyl- and (S)-3-hydroxybutyryl-moiety binding mode of enzyme Ms3HPCD. The residues involved in the formation of the 3-hydroxypropanoate binding pocket are identified. Ms3HPCD cannot convert (R)-stereoisomer of 3-hydroxybutyryl-CoA. When (R)-3-hydroxybutyryl-CoA is used as a substrate, the positions of the 3-hydroxyl-group and the C4-moiety are reversed each other, resulting in improper positioning of the (R)-3-hydroxybutyryl-moiety in the pocket. Ms3HPCD has a tightly formed alpha3 helix near the active site, and bulky aromatic residues are located at the enoyl-group binding site, resulting in the enzyme having an optimal substrate binding site for accepting short-chain 3-hydroxyacyl-CoA as a substrate
Ms3HPCD shows an overall structure and the CoA-binding mode similar to other enoyl-CoA hydratase (ECH) family enzymes, but compared with the other ECHs, Ms3HPCD has a tightly formed alpha3 helix near the active site, and bulky aromatic residues are located at the enoyl-group binding site, resulting in the enzyme having an optimal substrate binding site for accepting short chain 3-hydroxyacyl-CoA as a substrate. Phylogenetic tree analysis. The 3HPCD homologues from the phylum Crenarchaeota have an enoyl-group binding pocket similar to that of bacterial short-chain ECHs
Ms3HPCD shows an overall structure and the CoA-binding mode similar to other enoyl-CoA hydratase (ECH) family enzymes, but compared with the other ECHs, Ms3HPCD has a tightly formed alpha3 helix near the active site, and bulky aromatic residues are located at the enoyl-group binding site, resulting in the enzyme having an optimal substrate binding site for accepting short chain 3-hydroxyacyl-CoA as a substrate. Phylogenetic tree analysis. The 3HPCD homologues from the phylum Crenarchaeota have an enoyl-group binding pocket similar to that of bacterial short-chain ECHs
3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) cycle fixes CO2 in extremely thermoacidophilic archaea and holds promise for metabolic engineering because of its thermostability and potentially rapid pathway kinetics. Modelling of the reactions and kinetics of five of the cycle enzymes: malonyl-CoA/succinyl-CoA reductase, 3-hydroxypropionyl-CoA synthetase, 3-hydroxypropionyl-CoA dehydratase, acryloyl-CoA reductase, and succinic semialdehyde reductase, that are recombinantly expressed in Escherichia coli. Reaction kinetics model of the 3HP/4HB cycle, overview
Metallosphaera sedula is a thermoacidophilic autotrophic archaeon known to utilize the 3-hydroxypropionate/4-hydroxybutyrate cycle (3-HP/4-HB cycle) as carbon fixation pathway. 3-Hydroxypropionyl-CoA dehydratase (3HPCD) is an enzyme involved in the 3-HP/4-HB cycle by converting 3-hydroxypropionyl-CoA to acryloyl-CoA
3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) cycle fixes CO2 in extremely thermoacidophilic archaea and holds promise for metabolic engineering because of its thermostability and potentially rapid pathway kinetics. Modelling of the reactions and kinetics of five of the cycle enzymes: malonyl-CoA/succinyl-CoA reductase, 3-hydroxypropionyl-CoA synthetase, 3-hydroxypropionyl-CoA dehydratase, acryloyl-CoA reductase, and succinic semialdehyde reductase, that are recombinantly expressed in Escherichia coli. Reaction kinetics model of the 3HP/4HB cycle, overview
Metallosphaera sedula is a thermoacidophilic autotrophic archaeon known to utilize the 3-hydroxypropionate/4-hydroxybutyrate cycle (3-HP/4-HB cycle) as carbon fixation pathway. 3-Hydroxypropionyl-CoA dehydratase (3HPCD) is an enzyme involved in the 3-HP/4-HB cycle by converting 3-hydroxypropionyl-CoA to acryloyl-CoA
molecular docking simulations of 3-hydroxypropanoyl-CoA and (S)-3-hydroxybutyryl-CoA to Ms3HPCD structure, overview. When (R)-3-hydroxybutyryl-CoA is used as a substrate, the positions of the 3-hydroxyl-group and the C4-moiety are reversed each other, resulting in improper positioning of the (R)-3-hydroxybutyryl-moiety in the pocket. Structural comparison of Ms3HPCD with other enoyl-CoA hydratases, Structural basis for 3-hydroxypropanoyl-CoA substrate specificity of Ms3HPCD and active site structure, overview. Glutamate residues, Glu113 and Glu133, which act as catalytic acid and base, respectively, are positioned at the active site of Ms3HPCD
molecular docking simulations of 3-hydroxypropanoyl-CoA and (S)-3-hydroxybutyryl-CoA to Ms3HPCD structure, overview. When (R)-3-hydroxybutyryl-CoA is used as a substrate, the positions of the 3-hydroxyl-group and the C4-moiety are reversed each other, resulting in improper positioning of the (R)-3-hydroxybutyryl-moiety in the pocket. Structural comparison of Ms3HPCD with other enoyl-CoA hydratases, Structural basis for 3-hydroxypropanoyl-CoA substrate specificity of Ms3HPCD and active site structure, overview. Glutamate residues, Glu113 and Glu133, which act as catalytic acid and base, respectively, are positioned at the active site of Ms3HPCD
yeast two-hybrid assay for protein interaction analysis of 3-hydroxypropionyl-CoA dehydratase and 3-hydroxypropionyl-CoA synthetase (HPCS-HPCD), method optimization, metabolic engineering, overview
yeast two-hybrid assay for protein interaction analysis of 3-hydroxypropionyl-CoA dehydratase and 3-hydroxypropionyl-CoA synthetase (HPCS-HPCD), method optimization, metabolic engineering, overview
yeast two-hybrid assay for protein interaction analysis of 3-hydroxypropionyl-CoA dehydratase and 3-hydroxypropionyl-CoA synthetase (HPCS-HPCD), method optimization, metabolic engineering, overview
molecular docking simulations of 3-hydroxypropanoyl-CoA and (S)-3-hydroxybutyryl-CoA to Ms3HPCD structure, overview. When (R)-3-hydroxybutyryl-CoA is used as a substrate, the positions of the 3-hydroxyl-group and the C4-moiety are reversed each other, resulting in improper positioning of the (R)-3-hydroxybutyryl-moiety in the pocket. Structural comparison of Ms3HPCD with other enoyl-CoA hydratases, Structural basis for 3-hydroxypropanoyl-CoA substrate specificity of Ms3HPCD and active site structure, overview. Glutamate residues, Glu113 and Glu133, which act as catalytic acid and base, respectively, are positioned at the active site of Ms3HPCD
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified enzyme in complex with CoA, hanging drop vapor diffusion method, mixing of 0.001 ml of protein solution containing 41 mg/ml in 40 mM Tris-HCl, pH8.0, with 0.001 ml of reservoir solution, containing consisting of 10% w/v PEG 8000, 0.1 M sodium-potassium phosphate, pH 6.2, 0.2 M NaCl, and 10 mM EDTA, and equilibration against 0.05 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.8 A resolution, modelling
production of the commercially promising platform chemical 3-hydroxypropionic acid (3-HP) via the propionyl-CoA pathway in genetically engineered Escherichia coli strain BL21(DE3). Propionate CoA-transferase from Megasphaera elsdenii and 3-hydroxypropionyl-CoA dehydratase (HPCD) from Chloroflexus aurantiacus are expressed along with propionyl-CoA dehydrogenase (PACD) from Candida rugosa, the 3-hydroxypropanoate titer of the resulting Escherichia coli Ec-PPH strain is improved by 6fold. When cultured at 30°C with 1% glucose in addition to propionate, 3-hydroxypropanoate production by Ec-PPH increases 2fold and 12fold compared to the cultivation at 37°C (4.23 mM) or without glucose (0.68 mM), respectively. Deletion of the ygfH gene encoding propionyl-CoA: succinate CoA-transferase from Ec-PPH (resulting in the strain Ec-DELTAY-PPH) leads to increase of 3-hydroxypropanoate production in shake flask experiments (15.04 mM), whereas the strain Ec-DELTAY-PPH with deletion of the prpC gene (encoding methylcitrate synthase in the methylcitrate cycle) produces 17.76 mM of 3-HP. The strain Ec-DELTAY-DELTAP-PPH with both ygfH and prpC genes deleted produces 24.14 mM of 3-HP, thus showing an 18fold increase in the 3-hydroxypropanoate titer in compare to the strain Ec-P. Disruption of the competing metabolic pathways. Established transgenic metabolic pathway, method, overview
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21 (DE3)-T1 by nickel affinity chromatography and gel filtration to homogeneity
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene hpcd, coexpression of 3-hydroxypropionyl-CoA dehydratase (HPCD) with propionyl-CoA dehydrogenase (PACD) encoded by gene pacd from Candida rugosa and propionate CoA-transferase (PCT) encoded by gene pct from Megasphaera elsdenii in Escherichia coli strain BL21(DE3) under control of the T7 promoter
gene hpcd, recombinant expression of His-tagged enzyme HPCD in Escherichi coli strain Rosetta 2 (DE3), coexpression with 3-hydroxypropionyl-CoA synthetase, acryloyl-CoA reductase, and succinic semialdehyde reductase. Yeast two-hybrid assay for protein interaction analysis of 3-hydroxypropionyl-CoA dehydratase and 3-hydroxypropionyl-CoA synthetase (HPCS-HPCD)
gene Msed_2001, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic tree analysis, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21 (DE3)-T1
Teufel, R.; Kung, J.; Kockelkorn, D.; Alber, B.; Fuchs, G.
3-Hydroxypropionyl-coenzyme A dehydratase and acryloyl-coenzyme A reductase, enzymes of the autotrophic 3-hydroxypropionate/4-hydroxybutyrate cycle in the Sulfolobales