Literature summary for 3.5.2.2 extracted from

Cai, Y.; Trodler, P.; Jiang, S.; Zhang, W.; Wu, Y.; Lu, Y.; Yang, S.; Jiang, W.
Isolation and molecular characterization of a novel D-hydantoinase from Jannaschia sp. CCS1 (2009), FEBS J., 276, 3575-3588.

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli, His-tag	Jannaschia sp. CCS1

Crystallization (Commentary)

Crystallization (Comment)	Organism
docking of the productive transition states of hydantoin, DL-p-hydroxyphenylhydantoin, 1-p-hydroxyphenylhydantoin and dihydrouracil into the active site to simulate the mode of substrate binding. The substrate binding pocket accommodates substrates with small side chains better than those with large ones. The bulky side chains of Phe63, Leu92, Phe150 and Tyr153 are in close contact with the exocyclic substituent of D-p-hydroxyphenylhydantoin	Jannaschia sp. CCS1

Crystallization (Comment)

Organism

docking of the productive transition states of hydantoin, DL-p-hydroxyphenylhydantoin, 1-p-hydroxyphenylhydantoin and dihydrouracil into the active site to simulate the mode of substrate binding. The substrate binding pocket accommodates substrates with small side chains better than those with large ones. The bulky side chains of Phe63, Leu92, Phe150 and Tyr153 are in close contact with the exocyclic substituent of D-p-hydroxyphenylhydantoin

Jannaschia sp. CCS1

Protein Variants

Protein Variants	Comment	Organism
F150C	about 5% of wild-type activity	Jannaschia sp. CCS1
F150D	about 5% of wild-type activity	Jannaschia sp. CCS1
F150E	about 5% of wild-type activity	Jannaschia sp. CCS1
F150G	about 8% of wild-type activity	Jannaschia sp. CCS1
F150H	about 5% of wild-type activity	Jannaschia sp. CCS1
F150I	about 10% of wild-type activity	Jannaschia sp. CCS1
F150K	about 5% of wild-type activity	Jannaschia sp. CCS1
F150L	about 10% of wild-type activity	Jannaschia sp. CCS1
F150M	about 15% of wild-type activity	Jannaschia sp. CCS1
F150N	about 5% of wild-type activity	Jannaschia sp. CCS1
F150P	about 5% of wild-type activity	Jannaschia sp. CCS1
F150Q	about 5% of wild-type activity	Jannaschia sp. CCS1
F150R	about 5% of wild-type activity	Jannaschia sp. CCS1
F150S	about 5% of wild-type activity	Jannaschia sp. CCS1
F150T	about 5% of wild-type activity	Jannaschia sp. CCS1
F150V	about 5% of wild-type activity	Jannaschia sp. CCS1
F150W	about 5% of wild-type activity	Jannaschia sp. CCS1
F150Y	about 18% of wild-type activity	Jannaschia sp. CCS1
F63C	about 75% of wild-type activity	Jannaschia sp. CCS1
F63D	about 15% of wild-type activity	Jannaschia sp. CCS1
F63E	about 10% of wild-type activity	Jannaschia sp. CCS1
F63G	about 50% of wild-type activity	Jannaschia sp. CCS1
F63H	about 115% of wild-type activity	Jannaschia sp. CCS1
F63I	about 80% of wild-type activity	Jannaschia sp. CCS1
F63K	about 35% of wild-type activity	Jannaschia sp. CCS1
F63L	about 105% of wild-type activity	Jannaschia sp. CCS1
F63M	about 95% of wild-type activity	Jannaschia sp. CCS1
F63N	about 110% of wild-type activity	Jannaschia sp. CCS1
F63P	about 10% of wild-type activity	Jannaschia sp. CCS1
F63Q	about 25% of wild-type activity	Jannaschia sp. CCS1
F63R	about 30% of wild-type activity	Jannaschia sp. CCS1
F63S	about 75% of wild-type activity	Jannaschia sp. CCS1
F63T	about 70% of wild-type activity	Jannaschia sp. CCS1
F63V	about 70% of wild-type activity	Jannaschia sp. CCS1
F63W	about 95% of wild-type activity	Jannaschia sp. CCS1
F63Y	about 125% of wild-type activity	Jannaschia sp. CCS1
L157A	33.6% of wild-type activity	Jannaschia sp. CCS1
L157D	11% of wild-type activity	Jannaschia sp. CCS1
L157I	94.0% of wild-type activity	Jannaschia sp. CCS1
L157V	98.3% of wild-type activity	Jannaschia sp. CCS1
L92C	about 25% of wild-type activity	Jannaschia sp. CCS1
L92D	about 10% of wild-type activity	Jannaschia sp. CCS1
L92E	about 10% of wild-type activity	Jannaschia sp. CCS1
L92F	about 95% of wild-type activity	Jannaschia sp. CCS1
L92G	about 10% of wild-type activity	Jannaschia sp. CCS1
L92H	about 15% of wild-type activity	Jannaschia sp. CCS1
L92I	about 120% of wild-type activity	Jannaschia sp. CCS1
L92K	about 20% of wild-type activity	Jannaschia sp. CCS1
L92M	about 115% of wild-type activity	Jannaschia sp. CCS1
L92N	about 10% of wild-type activity	Jannaschia sp. CCS1
L92P	about 5% of wild-type activity	Jannaschia sp. CCS1
L92Q	about 15% of wild-type activity	Jannaschia sp. CCS1
L92R	about 5% of wild-type activity	Jannaschia sp. CCS1
L92S	about 10% of wild-type activity	Jannaschia sp. CCS1
L92T	about 35% of wild-type activity	Jannaschia sp. CCS1
L92V	about 70% of wild-type activity	Jannaschia sp. CCS1
L92W	about 10% of wild-type activity	Jannaschia sp. CCS1
L92Y	about 5% of wild-type activity	Jannaschia sp. CCS1

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
18	-	DL-p-hydroxyphenylhydantoin	pH 8.0, 40°C	Jannaschia sp. CCS1

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
56000	-	4 * 56000, His-tagged recombinant protein	Jannaschia sp. CCS1
253000	-	gel filtration	Jannaschia sp. CCS1

Organism

Organism	UniProt	Comment	Textmining
Jannaschia sp. CCS1	Q28SA7	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant protein	Jannaschia sp. CCS1

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
8.2	-	40°C, pH 8.0	Jannaschia sp. CCS1

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
5,6-dihydrouracil + H2O	-	Jannaschia sp. CCS1	3-ureidopropionate	-	?
dimethylhydantoin + H2O	1.4% of the activity with dihydrouracil	Jannaschia sp. CCS1	?	-	?
DL-5-phenylhydantoin + H2O	45% of the activity with dihydrouracil	Jannaschia sp. CCS1	N-carbamoyl-D-phenylglycine	-	?
DL-p-hydroxyphenylhydantoin + H2O	7.2% of the activity with dihydrouracil	Jannaschia sp. CCS1	N-carbamoyl-D-p-hydroxyphenylglycine	-	?
hydantoin + H2O	18.7% of the activity with dihydrouracil	Jannaschia sp. CCS1	hydantoic acid	-	?
additional information	no substrate: diphenylhydantoin, 5-(hydroxymethyl)uracil, benzylhydantoin, isopropylhydantoin	Jannaschia sp. CCS1	?	-	?

Subunits

Subunits	Comment	Organism
tetramer	4 * 56000, His-tagged recombinant protein	Jannaschia sp. CCS1

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	-	Jannaschia sp. CCS1

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
4.6	-	DL-p-hydroxyphenylhydantoin	pH 8.0, 40°C	Jannaschia sp. CCS1

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.6	-	-	Jannaschia sp. CCS1

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.25	-	DL-p-hydroxyphenylhydantoin	pH 8.0, 40°C	Jannaschia sp. CCS1