Cloned (Comment) | Organism |
---|---|
recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) pLysS | Advenella mimigardefordensis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant apoenzyme and enzyme in complex with the CoA moiety from the substrate analogue succinyl-CoA, hanging drop vapour diffusion, mixing of 0.002 ml of 10 mg/ml protein in 50 mM Tris-HCl, pH 7.4, with 0.002 ml of reservoir solution containing 0.1 M Bis-Tris, pH 6.5, and 5-20% PEG 3350, in the absence or presence of 4 mM succinyl-CoA, X-ray diffraction structure determination and analysis at 1.89 A and 2.30 A resolution, respectively, molecular replacement using the monomer of an acyl-CoA dehydrogenase (Acd) from Thermus thermophilus HB8 as search model | Advenella mimigardefordensis |
Protein Variants | Comment | Organism |
---|---|---|
Q246E | site-directed mutagenesis, substrate specificity of the mutant | Advenella mimigardefordensis |
R84K | site-directed mutagenesis, the mutant shows a complete loss of enzymatic activity | Advenella mimigardefordensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
3-sulfinopropanoyl-CoA | recombinant His6-tagged wild-type enzyme, pH 7.4, 30°C | Advenella mimigardefordensis | |
0.019 | - |
3-sulfinopropanoyl-CoA | recombinant His6-tagged mutant Q246E, pH 7.4, 30°C | Advenella mimigardefordensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-sulfinopropanoyl-CoA + H2O | Advenella mimigardefordensis | - |
propanoyl-CoA + sulfite | - |
? | |
3-sulfinopropanoyl-CoA + H2O | Advenella mimigardefordensis DPN7T | - |
propanoyl-CoA + sulfite | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Advenella mimigardefordensis | K4L7X3 | - |
- |
Advenella mimigardefordensis DPN7T | K4L7X3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) pLysS by nicke affinity chromatography and gel filtration | Advenella mimigardefordensis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
3-sulfinopropanoyl-CoA + H2O = propanoyl-CoA + sulfite | reaction mechanism, residues Arg84, Asp88 and Gln246 side chains and the isoalloxazine ring of FAD define the enzymatic environment for the desulfination reaction. FAD is not involved in electron transfer during the reaction, essential role of Arg84 in catalysis. The enzymatic reaction might progress with hydrogen-bond formation and charge transfer between the guanidinium cation of Arg84 and 3-sulfinopropanoyl-CoA, which would favour nucleophilic attack of a water molecule on the S atom of the sulfino group | Advenella mimigardefordensis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4.19 | - |
purified recombinant His6-tagged wild-type enzyme, pH 7.4, 30°C | Advenella mimigardefordensis |
5.26 | - |
purified recombinant His6-tagged mutant Q246E, pH 7.4, 30°C | Advenella mimigardefordensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-sulfinopropanoyl-CoA + H2O | - |
Advenella mimigardefordensis | propanoyl-CoA + sulfite | - |
? | |
3-sulfinopropanoyl-CoA + H2O | - |
Advenella mimigardefordensis DPN7T | propanoyl-CoA + sulfite | - |
? | |
additional information | the enzyme does not show any dehydrogenase activity | Advenella mimigardefordensis | ? | - |
? | |
additional information | the enzyme does not show any dehydrogenase activity | Advenella mimigardefordensis DPN7T | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | - |
Advenella mimigardefordensis |
Synonyms | Comment | Organism |
---|---|---|
3-sulfinopropionyl-coenzyme A desulfinase | - |
Advenella mimigardefordensis |
3SP-CoA desulfinase | - |
Advenella mimigardefordensis |
AcdDPN7 | - |
Advenella mimigardefordensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Advenella mimigardefordensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.13 | - |
3-sulfinopropanoyl-CoA | recombinant His6-tagged wild-type enzyme, pH 7.4, 30°C | Advenella mimigardefordensis | |
3.93 | - |
3-sulfinopropanoyl-CoA | recombinant His6-tagged mutant Q246E, pH 7.4, 30°C | Advenella mimigardefordensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Advenella mimigardefordensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | the enzyme is a tetramer with each subunit containing one flavin adenine dinucleotide (FAD) molecule, binding structure, overview. FAD is not involved in electron transfer during the reaction | Advenella mimigardefordensis |
General Information | Comment | Organism |
---|---|---|
evolution | the apoenzyme structure shows that AcdDPN7 belongs to the acyl-CoA dehydrogenase superfamily fold | Advenella mimigardefordensis |
metabolism | the enzyme catalyzes the key step in the the 3,3'-dithiodipropionic acid (DTDP) catabolic pathway, overview | Advenella mimigardefordensis |
additional information | Arg84 is a key residue in the desulfination reaction, the guanidinium group of the arginine is essential for desulfination, modelling of the enzyme-substrate complex, overview. Structure-function analysis and active site structure determination, modeling of substrate binding. The Arg84, Asp88 and Gln246 side chains and the isoalloxazine ring of FAD define the enzymatic environment for the desulfination reaction | Advenella mimigardefordensis |
physiological function | the enzyme catalyzes sulfur abstraction from 3-sulfinopropionyl-CoA, a key step during the catabolism of 3,3'-dithiodipropionic acid | Advenella mimigardefordensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
203.2 | - |
3-sulfinopropanoyl-CoA | recombinant His6-tagged mutant Q246E, pH 7.4, 30°C | Advenella mimigardefordensis | |
237.2 | - |
3-sulfinopropanoyl-CoA | recombinant His6-tagged wild-type enzyme, pH 7.4, 30°C | Advenella mimigardefordensis |