Literature summary for 4.2.1.1 extracted from

De Simone, G.; Monti, S.M.; Alterio, V.; Buonanno, M.; De Luca, V.; Rossi, M.; Carginale, V.; Supuran, C.T.; Capasso, C.; Di Fiore, A.
Crystal structure of the most catalytically effective carbonic anhydrase enzyme known, SazCA from the thermophilic bacterium Sulfurihydrogenibium azorense (2015), Bioorg. Med. Chem. Lett., 25, 2002-2006 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Sulfurihydrogenibium azorense

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme incomplex with inhibitor acetazolamide, hanging drop vapor diffusion method, mixing of 0.001 ml of 8 mg/ml protein in 20 mM Tris, pH 8.3, and a 10-molar excess of the inhibitor with 0.001 m of reservoir solution containing 30% w/v PEG monomethyl ether 550, 0.05 M MgCl2, and 0.1 M HEPES, pH 7.5, and equilibration against 0.5 ml of reservoir solution, 1 week, room temperature, X-ray diffraction structure determination and analysis at 1.95 A resolution, molecular replacement using the structure of enzyme SspCA from Sulfurihydrogenibium yellowstonensis (PDB ID 4G7A) as a search model	Sulfurihydrogenibium azorense

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme incomplex with inhibitor acetazolamide, hanging drop vapor diffusion method, mixing of 0.001 ml of 8 mg/ml protein in 20 mM Tris, pH 8.3, and a 10-molar excess of the inhibitor with 0.001 m of reservoir solution containing 30% w/v PEG monomethyl ether 550, 0.05 M MgCl2, and 0.1 M HEPES, pH 7.5, and equilibration against 0.5 ml of reservoir solution, 1 week, room temperature, X-ray diffraction structure determination and analysis at 1.95 A resolution, molecular replacement using the structure of enzyme SspCA from Sulfurihydrogenibium yellowstonensis (PDB ID 4G7A) as a search model

Sulfurihydrogenibium azorense

Inhibitors

Inhibitors	Comment	Organism	Structure
acetazolamide	AZM, strong inhibitor	Sulfurihydrogenibium azorense

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	required, metalloenzyme, on the bottom of a central cavity, the catalytic zinc ion is tetrahedrally coordinated by the three histidine residues (His89, His91 and His108)	Sulfurihydrogenibium azorense

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
CO2 + H2O	Sulfurihydrogenibium azorense	-	H2CO3	-	r
CO2 + H2O	Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	-	H2CO3	-	r
H2CO3	Sulfurihydrogenibium azorense	-	CO2 + H2O	-	r
H2CO3	Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	-	CO2 + H2O	-	r
additional information	Sulfurihydrogenibium azorense	enzyme SazCA is highly active in the CO2 hydration reaction	?	-	?
additional information	Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	enzyme SazCA is highly active in the CO2 hydration reaction	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Sulfurihydrogenibium azorense	C1DTU5	-	-
Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	C1DTU5	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and gel filtration, followed by pAMBS affinity chromatography	Sulfurihydrogenibium azorense

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
CO2 + H2O	-	Sulfurihydrogenibium azorense	H2CO3	-	r
CO2 + H2O	-	Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	H2CO3	-	r
H2CO3	-	Sulfurihydrogenibium azorense	CO2 + H2O	-	r
H2CO3	-	Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	CO2 + H2O	-	r
additional information	enzyme SazCA is highly active in the CO2 hydration reaction	Sulfurihydrogenibium azorense	?	-	?
additional information	enzyme SazCA is highly active in the CO2 hydration reaction	Sulfurihydrogenibium azorense Az-Fu1 / DSM 15241 / OCM 825	?	-	?

Subunits

Subunits	Comment	Organism
homodimer	SazCA enzyme three-dimensional structure determination and analysis, structure comparison with the enzyme from Sulfurihydrogenibium yellowstonensis, PDB ID 4G7A. The SazCA monomer adopts the canonical fold of the alpha-CA family members consisting of a central ten-stranded beta-sheet surrounded by alpha- and 310-helices as well as small additional beta-sheets. The active site of each monomer within the dimer is completely accessible to the solvent and, as observed for other alpha-CAs, is located in a cavity which extends from the surface to the center of the protein	Sulfurihydrogenibium azorense

Synonyms

Synonyms	Comment	Organism
alpha-CA	-	Sulfurihydrogenibium azorense
alpha-carbonic anhydrase	-	Sulfurihydrogenibium azorense
SazCA	-	Sulfurihydrogenibium azorense
SULAZ_0541	-	Sulfurihydrogenibium azorense

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
20	-	assay at	Sulfurihydrogenibium azorense

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
4400000	-	CO2	pH 7.5, 20°C, recombinant His-tagged enzyme	Sulfurihydrogenibium azorense

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Sulfurihydrogenibium azorense

General Information

General Information	Comment	Organism
additional information	identification of the structural features responsible for the high catalytic activity of the enzyme from Sulfurihydrogenibium azorense, structure comparison with the enzyme from Sulfurihydrogenibium yellowstonensis. The active site of each monomer within the dimer is completely accessible to the solvent and, as observed for other alpha-CAs, is located in a cavity which extends from the surface to the center of the protein. On the bottom of this cavity, the catalytic zinc ion is tetrahedrally coordinated by the three histidine residues (His89, His91 and His108). Enzyme SazCA active site structure and environment, important for catalytic activity, overview	Sulfurihydrogenibium azorense

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
350000	-	CO2	pH 7.5, 20°C, recombinant His-tagged enzyme	Sulfurihydrogenibium azorense