Literature summary for 4.2.1.8 extracted from

Zhang, Q.; Gao, F.; Peng, H.; Cheng, H.; Liu, Y.; Tang, J.; Thompson, J.; Wei, G.; Zhang, J.; Du, Y.; Yan, J.; Gao, G.F.
Crystal structures of Streptococcus suis mannonate dehydratase (ManD) and its complex with substrate: genetic and biochemical evidence for a catalytic mechanism (2009), J. Bacteriol., 191, 5832-5837.

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Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Streptococcus suis

Crystallization (Commentary)

Crystallization (Comment)	Organism
ManD in native form and in complex with its substrate D-mannonate and Mn2+ ion, hanging drop vapor diffusion method, using 0.2 M potassium-sodium tartrate, 0.1 M sodium citrate (pH 6.5), and 1 M ammonium sulfate (native form) or using 0.2 M potassium sodium tartrate tetrahydrate, 0.1 M trisodium citrate dehydrate (pH 6.5), 1 M ammonium sulfate, and 15 mM D-mannonate	Streptococcus suis

Protein Variants

Protein Variants	Comment	Organism
H311A	the activity of H311A is only 2.4% that of the native wild type enzyme	Streptococcus suis
Y325F	catalytically inactive	Streptococcus suis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
1.9	-	D-mannonate	mutant enzyme H311A, at pH 7.5 and 37°C	Streptococcus suis
3	-	D-mannonate	wild type enzyme, at pH 7.5	Streptococcus suis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	each ManD monomer roughly has one Mn2+ ion-binding site	Streptococcus suis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
41000	-	2 * 41000, SDS-PAGE	Streptococcus suis
80000	-	gel filtration	Streptococcus suis

Organism

Organism	UniProt	Comment	Textmining
Streptococcus suis	A4VVI4	serotype 2	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-NTA column chromatography, Superdex 200 gel filtration	Streptococcus suis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Mannonate	-	Streptococcus suis	2-Dehydro-3-deoxy-D-gluconate + H2O	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * 41000, SDS-PAGE	Streptococcus suis

Synonyms

Synonyms	Comment	Organism
ManD	-	Streptococcus suis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
4	-	ManD exhibits no enzymatic activity at 4°C	Streptococcus suis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.09	-	D-mannonate	mutant enzyme H311A, at pH 7.5 and 37°C	Streptococcus suis
5.88	-	D-mannonate	wild type enzyme, at pH 7.5 and 37°C	Streptococcus suis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.047	-	D-mannonate	mutant enzyme H311A, at pH 7.5 and 37°C	Streptococcus suis
1.96	-	D-mannonate	wild type enzyme, at pH 7.5 and 37°C	Streptococcus suis

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Release 2023.1 (January 2023)