Literature summary for 3.2.1.26 extracted from

Alvaro-Benito, M.; Polo, A.; Gonzalez, B.; Fernandez-Lobato, M.; Sanz-Aparicio, J.
Structural and kinetic analysis of Schwanniomyces occidentalis invertase reveals a new oligomerization pattern and the role of its supplementary domain in substrate binding (2010), J. Biol. Chem., 285, 13930-13941.

Crystallization (Commentary)

Crystallization (Comment)	Organism
three-dimensional structure of invertase at 2.9 A resolution and its complex with fructose at 1.9 A resolution. The monomer presents a bimodular arrangement common to other GH32 enzymes, with an N-terminal 5fold beta-propeller catalytic domain and a C-terminal alpha-sandwich domain. Structure displays an unique active site cleft shaped by both subunits. Medium size and long substrates are recognized by residues from both subunits	Schwanniomyces occidentalis

Crystallization (Comment)

Organism

three-dimensional structure of invertase at 2.9 A resolution and its complex with fructose at 1.9 A resolution. The monomer presents a bimodular arrangement common to other GH32 enzymes, with an N-terminal 5fold beta-propeller catalytic domain and a C-terminal alpha-sandwich domain. Structure displays an unique active site cleft shaped by both subunits. Medium size and long substrates are recognized by residues from both subunits

Schwanniomyces occidentalis

Protein Variants

Protein Variants	Comment	Organism
D50A	no catalytic activity	Schwanniomyces occidentalis
E230A	no catalytic activity	Schwanniomyces occidentalis
N142Y	large decrease in activity against nystose	Schwanniomyces occidentalis
N254A	large decrease in activity against nystose	Schwanniomyces occidentalis
Q176E	moderate change in catalytic activity	Schwanniomyces occidentalis
Q176S	moderate change in catalytic activity	Schwanniomyces occidentalis
Q228V	strikingly large reduction in the catalytic efficiency of sucrose , inulin and especially nystose	Schwanniomyces occidentalis
Q435A	significant decrease in catalytic efficiency against inulin	Schwanniomyces occidentalis
S281I	strikingly large reduction in the catalytic efficiency of sucrose , inulin and especially nystose	Schwanniomyces occidentalis
Y462A	significant decrease in catalytic efficiency against inulin	Schwanniomyces occidentalis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
2.4	-	sucrose	wild-type, pH 5.0	Schwanniomyces occidentalis
3.8	-	nystose	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
5	-	sucrose	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
6.4	-	sucrose	wild-type, pH 5.0	Schwanniomyces occidentalis
7	-	inulin	mutant N254A, pH 5.0	Schwanniomyces occidentalis
7	-	sucrose	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
7	-	sucrose	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
8	-	nystose	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
8	-	sucrose	mutant N254A, pH 5.0	Schwanniomyces occidentalis
8	-	nystose	mutant N254A, pH 5.0	Schwanniomyces occidentalis
8	-	nystose	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
8.1	-	nystose	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
9	-	nystose	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
9	-	sucrose	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
9	-	inulin	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
11	-	nystose	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
11	-	sucrose	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
12	-	inulin	wild-type, pH 5.0	Schwanniomyces occidentalis
13	-	sucrose	mutant S281I, pH 5.0	Schwanniomyces occidentalis
14	-	inulin	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
15	-	nystose	mutant S281I, pH 5.0	Schwanniomyces occidentalis
24	-	inulin	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
29	-	sucrose	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
44	-	inulin	mutant S281I, pH 5.0	Schwanniomyces occidentalis
47	-	inulin	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
60	-	inulin	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
575	-	inulin	mutant Q176S, pH 5.0	Schwanniomyces occidentalis

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	-	Schwanniomyces occidentalis	-	-

Organism

Organism	UniProt	Comment	Textmining
Schwanniomyces occidentalis	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
inulin + H2O	-	Schwanniomyces occidentalis	D-fructose + ?	-	?
nystose + H2O	-	Schwanniomyces occidentalis	D-fructose + D-glucose	-	?
sucrose + H2O	-	Schwanniomyces occidentalis	D-fructose + D-glucose	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
2	-	nystose	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
2	-	inulin	mutant S281I, pH 5.0	Schwanniomyces occidentalis
3	-	inulin	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
5	-	inulin	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
6	-	nystose	mutant S281I, pH 5.0	Schwanniomyces occidentalis
10	-	inulin	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
12	-	inulin	mutant N254A, pH 5.0	Schwanniomyces occidentalis
20	-	sucrose	mutant S281I, pH 5.0	Schwanniomyces occidentalis
22	-	nystose	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
25	-	inulin	wild-type, pH 5.0	Schwanniomyces occidentalis
33	-	sucrose	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
39	-	inulin	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
43	-	sucrose	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
60	-	nystose	mutant N254A, pH 5.0	Schwanniomyces occidentalis
68	-	inulin	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
76	-	nystose	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
84	-	sucrose	wild-type, pH 5.0	Schwanniomyces occidentalis
85	-	sucrose	mutant N254A, pH 5.0	Schwanniomyces occidentalis
87	-	nystose	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
88	-	sucrose	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
92	-	nystose	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
99	-	sucrose	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
106	-	sucrose	wild-type, pH 5.0	Schwanniomyces occidentalis
134	-	sucrose	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
230	-	nystose	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
262	-	sucrose	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
1710	-	inulin	mutant Q176S, pH 5.0	Schwanniomyces occidentalis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.04	-	inulin	mutant S281I, pH 5.0	Schwanniomyces occidentalis
0.06	-	inulin	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
0.2	-	nystose	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
0.4	-	nystose	mutant S281I, pH 5.0	Schwanniomyces occidentalis
0.5	-	inulin	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
0.7	-	inulin	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
1.1	-	inulin	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
1.5	-	sucrose	mutant S281I, pH 5.0	Schwanniomyces occidentalis
1.6	-	inulin	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
1.7	-	inulin	mutant N254A, pH 5.0	Schwanniomyces occidentalis
2.1	-	inulin	wild-type, pH 5.0	Schwanniomyces occidentalis
2.7	-	nystose	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
3	-	inulin	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
3	-	sucrose	mutant Q228V, pH 5.0	Schwanniomyces occidentalis
6	-	sucrose	mutant Y462A, pH 5.0	Schwanniomyces occidentalis
7.5	-	nystose	mutant N254A, pH 5.0	Schwanniomyces occidentalis
9	-	sucrose	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
10.6	-	sucrose	mutant N254A, pH 5.0	Schwanniomyces occidentalis
11	-	nystose	mutant N142Y, pH 5.0	Schwanniomyces occidentalis
11	-	sucrose	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
12.6	-	sucrose	mutant Q176E, pH 5.0	Schwanniomyces occidentalis
16.6	-	sucrose	wild-type, pH 5.0	Schwanniomyces occidentalis
20	-	nystose	mutant Q435A, pH 5.0	Schwanniomyces occidentalis
21	-	nystose	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
26.8	-	sucrose	mutant Q176S, pH 5.0	Schwanniomyces occidentalis
35	-	sucrose	wild-type, pH 5.0	Schwanniomyces occidentalis
51	-	nystose	mutant Q176E, pH 5.0	Schwanniomyces occidentalis