Literature summary for 3.13.1.4 extracted from

Schuermann, M.; Meijers, R.; Schneider, T.R.; Steinbuechel, A.; Cianci, M.
3-Sulfinopropionyl-coenzyme A (3SP-CoA) desulfinase from Advenella mimigardefordensis DPN7(T): crystal structure and function of a desulfinase with an acyl-CoA dehydrogenase fold (2015), Acta Crystallogr. Sect. D, 71, 1360-1372.

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) pLysS	Advenella mimigardefordensis

Crystallization (Commentary)

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

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

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 [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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Advenella mimigardefordensis	K4L7X3	-	-
Advenella mimigardefordensis DPN7T	K4L7X3	-	-

Purification (Commentary)

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

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	a

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Subunits	Comment	Organism
tetramer	-	Advenella mimigardefordensis

Synonyms

Synonyms	Comment	Organism
3-sulfinopropionyl-coenzyme A desulfinase	-	Advenella mimigardefordensis
3SP-CoA desulfinase	-	Advenella mimigardefordensis
AcdDPN7	-	Advenella mimigardefordensis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Advenella mimigardefordensis

Turnover Number [1/s]

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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Advenella mimigardefordensis

Cofactor

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

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 [mM/s]

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