Literature summary for 3.2.1.22 extracted from

Fernandez-Leiro, R.; Pereira-Rodriguez, A.; Cerdan, M.E.; Becerra, M.; Sanz-Aparicio, J.
Structural analysis of Saccharomyces cerevisiae alpha-galactosidase and its complexes with natural substrates reveals new insights into substrate specificity of GH27 glycosidases (2010), J. Biol. Chem., 285, 28020-28033.

Search for more literature for 3.2.1.22

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structures of tetrameric Saccharomyces cerevisiae alpha-galactosidase and its complexes with the substrates melibiose and raffinose have been determined to 1.95, 2.40, and 2.70 A resolution. The monomer folds into a catalytic (alpha/beta)8 barrel and a C-terminal beta-sandwich domain with unassigned function	Saccharomyces cerevisiae

Protein Variants

Protein Variants	Comment	Organism
A41Y	kcat decreased compared to wild-tpye, Km (substrate: 4-nitrophenyl-alpha-D-galactopyranoside) or (substrate: melibiose) decreased compared to wild-type, Km (raffinose) increased compared to wild-type	Saccharomyces cerevisiae
D149A	mutant shows no activity	Saccharomyces cerevisiae
D209A	mutant shows no activity	Saccharomyces cerevisiae
N252A	mutant is highly unstable, no kinetic data available	Saccharomyces cerevisiae
Q251A	kcat and Km (substrate: 4-nitrophenyl-alpha-D-galactopyranoside) decreased compared to wild-type	Saccharomyces cerevisiae
Q251A	kcat decreased compared to wild-tpye, Km (substrate: 4-nitrophenyl-alpha-D-galactopyranoside) decreased compared to wild-type, Km (melibiose) increased compared to wild-type	Saccharomyces cerevisiae
Y232R	mutant is highly unstable, no kinetic data available	Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.24	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant Q251W, pH 7.4, 30°C	Saccharomyces cerevisiae
1.4	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant Q251A, pH 7.4, 30°C	Saccharomyces cerevisiae
3.7	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
4.5	-	4-nitrophenyl-alpha-D-galactopyranoside	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
5.8	-	melibiose	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
11.2	-	melibiose	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
54.1	-	raffinose	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
87.9	-	raffinose	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
254	-	melibiose	mutant Q251A, pH 7.4, 30°C	Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
80000	-	analytical ultracentrifugation under denaturing condition, monomer	Saccharomyces cerevisiae
320000	-	analytical ultracentrifugation under non-denaturing condition, tetramer	Saccharomyces cerevisiae

Organism

Organism	UniProt	Comment	Textmining
Saccharomyces cerevisiae	P04824	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl-alpha-D-galactopyranoside + H2O	-	Saccharomyces cerevisiae	4-nitrophenol + alpha-D-galactopyranose	-	?
melibiose + H2O	-	Saccharomyces cerevisiae	alpha-D-galactose + alpha-D-glucose	-	?
raffinose + H2O	-	Saccharomyces cerevisiae	sucrose + alpha-D-galactose	-	?

Subunits

Subunits	Comment	Organism
tetramer	analytical ultracentrifugation under non-denaturing condition, 4 * 80000 Da	Saccharomyces cerevisiae

Synonyms

Synonyms	Comment	Organism
alpha-galactosidase	-	Saccharomyces cerevisiae
ScAGal	-	Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
16	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant Q251W, pH 7.4, 30°C	Saccharomyces cerevisiae
46.8	-	raffinose	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
90	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
91	-	raffinose	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
113	-	melibiose	mutant Q251A, pH 7.4, 30°C	Saccharomyces cerevisiae
131	-	melibiose	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
170	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant Q251A, pH 7.4, 30°C	Saccharomyces cerevisiae
193	-	melibiose	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
286	-	4-nitrophenyl-alpha-D-galactopyranoside	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Saccharomyces cerevisiae

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.45	-	melibiose	mutant Q251A, pH 7.4, 30°C	Saccharomyces cerevisiae
0.8	-	raffinose	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
0.9	-	raffinose	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
17	-	melibiose	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
22	-	melibiose	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
24	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant A41Y, pH 7.4, 30°C	Saccharomyces cerevisiae
63.5	-	4-nitrophenyl-alpha-D-galactopyranoside	wild-type, pH 7.4, 30°C	Saccharomyces cerevisiae
66	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant Q251W, pH 7.4, 30°C	Saccharomyces cerevisiae
121	-	4-nitrophenyl-alpha-D-galactopyranoside	mutant Q251A, pH 7.4, 30°C	Saccharomyces cerevisiae

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Release 2023.1 (January 2023)