Literature summary for 3.2.1.26 extracted from

Perez de los Santos, A.I.; Cayetano-Cruz, M.; Gutierrez-Anton, M.; Santiago-Hernandez, A.; Plascencia-Espinosa, M.; Farres, A.; Hidalgo-Lara, M.E.
Improvement of catalytical properties of two invertases highly tolerant to sucrose after expression in Pichia pastoris. Effect of glycosylation on enzyme properties (2016), Enzyme Microb. Technol., 83, 48-56 .

Application

Application	Comment	Organism
biofuel production	the expression of INVA and INVB from Zymomonas mobilis in Pichia pastoris yields new catalysts with improved catalytic properties, making them suitable candidates for ethanol production from cane molasses	Zymomonas mobilis
industry	the expression of INVA and INVB from Zymomonas mobilis in Pichia pastoris yields new catalysts with improved catalytic properties, making them suitable candidates for a number of industrial applications	Zymomonas mobilis

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Pichia pastoris under the control of the strong AOX1 promoter. The expression levels of INVA(AOX1) (1660 U/mg) and INVB(AOX1) (1993 U/mg) in Pichia pastoris are 9- and 7-fold higher than the expression levels observed for the native INVA and INVB proteins in Zymomonas mobilis. Glycosylation of INVA(AOX1) and INVB(AOX1) plays an important role in their thermal stability, catalytic efficiency, and tolerance to sucrose	Zymomonas mobilis

Cloned (Comment)

Organism

expression in Pichia pastoris under the control of the strong AOX1 promoter. The expression levels of INVA(AOX1) (1660 U/mg) and INVB(AOX1) (1993 U/mg) in Pichia pastoris are 9- and 7-fold higher than the expression levels observed for the native INVA and INVB proteins in Zymomonas mobilis. Glycosylation of INVA(AOX1) and INVB(AOX1) plays an important role in their thermal stability, catalytic efficiency, and tolerance to sucrose

Zymomonas mobilis

Inhibitors

Inhibitors	Comment	Organism
Cu2+	inhibits invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, native invertase INVB and invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
Hg2+	inhibits native invertase INVA, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, native invertase INVB and native invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
Ni2+	inhibits native invertase INVB and invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
sucrose	invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter and invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter are highly active at sucrose concentrations of up to 400 and 300 mM, respectively	Zymomonas mobilis
Zn2+	inhibits native invertase INVB and invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
41	-	sucrose	pH 5.0, 35°C, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
50	-	sucrose	pH 5.5, 40°C, deglycosylated invertase INVB	Zymomonas mobilis
50	-	sucrose	pH 5.5, 40°C, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
83	-	sucrose	pH 5.0, 35°C, deglycosylated invertase INVA	Zymomonas mobilis
98	-	sucrose	pH 5.5, 40°C, native invertase INVB	Zymomonas mobilis
110	-	sucrose	pH 5.0, 35°C, native invertase INVA	Zymomonas mobilis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	the invertase activity increases by 80% (INVAAOX1) and by 20% (INVBAOX1) in the presence of Mn2+ at 1 mM and 5 mM, respectively	Zymomonas mobilis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
46000	-	deglycosylated invertase INVB, SDS-PAGE	Zymomonas mobilis
46000	-	native invertase INVB, SDS-PAGE	Zymomonas mobilis
48000	-	invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE	Zymomonas mobilis
57000	-	deglycosylated invertase INVA, SDS-PAGE	Zymomonas mobilis
57000	-	native invertase INVA, SDS-PAGE	Zymomonas mobilis
68000	-	invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE	Zymomonas mobilis

Organism

Organism	UniProt	Comment	Textmining
Zymomonas mobilis	-	-	-
Zymomonas mobilis CDBB-B603	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	after deglycosylation of the enzymes, denoted D-INVAAOX1and D-INVBAOX1, they exhibit a 1.3- and 3-fold lower catalytic efficiency (107 and 164 s-1*mM-1, respectively), and a 1.3- to 5-fold lower thermal stability than the glycosylated forms at temperatures of 35-45°C. After deglycosylation no effect is observed in optimal pH, being of 5.5 for INVAAOX1, INVBAOX1, D-INVAAOX1 and D-INVBAOX1	Zymomonas mobilis

Purification (Commentary)

Purification (Comment)	Organism
-	Zymomonas mobilis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
sucrose	INVA(AOX1) and INVB(AOX1) are highly active at sucrose concentrations of up to 400 and 300 mM, respectively. The tolerance to sucrose decreases to 300 mM for D-INVA(AOX1)	Zymomonas mobilis	beta-D-fructose + alpha-D-glucose	-	?
sucrose	INVA(AOX1) and INVB(AOX1) are highly active at sucrose concentrations of up to 400 and 300 mM, respectively. The tolerance to sucrose decreases to 300 mM for D-INVA(AOX1)	Zymomonas mobilis CDBB-B603	beta-D-fructose + alpha-D-glucose	-	?

Subunits

Subunits	Comment	Organism
?	x * 46000, deglycosylated invertase INVB, SDS-PAGE	Zymomonas mobilis
?	x * 46000, native invertase INVB, SDS-PAGE	Zymomonas mobilis
?	x * 48000, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE	Zymomonas mobilis
?	x * 57000, deglycosylated invertase INVA, SDS-PAGE	Zymomonas mobilis
?	x * 57000, native invertase INVA, SDS-PAGE	Zymomonas mobilis
?	x * 68000, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE	Zymomonas mobilis

Synonyms

Synonyms	Comment	Organism
INVA	-	Zymomonas mobilis
INVB	-	Zymomonas mobilis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	native invertase INVA and deglycosylated invertase INVA	Zymomonas mobilis
35	-	assay at, invertase INVA	Zymomonas mobilis
35	-	invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
40	-	assay at, invertase INVB	Zymomonas mobilis
40	-	native invertase INVB, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter and deglycosylated invertase INVB	Zymomonas mobilis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
35	-	t1/2 of invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter is 900 min, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter is 80 min	Zymomonas mobilis
35	45	after deglycosylation of the enzymes, denoted D-INVAAOX1and D-INVBAOX1, they exhibit a 1.3- to 5-fold lower thermal stability than the glycosylated forms at temperatures of 35-45°C	Zymomonas mobilis
45	-	t1/2 of invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter is 480 min, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter is 50 min	Zymomonas mobilis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
56.92	-	sucrose	pH 5.0, 35°C, native invertase INVA	Zymomonas mobilis
8210	-	sucrose	pH 5.5, 40°C, deglycosylated invertase INVB	Zymomonas mobilis
8831	-	sucrose	pH 5.0, 35°C, deglycosylated invertase INVA	Zymomonas mobilis
10396	-	sucrose	pH 5.5, 40°C, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
12000	-	sucrose	pH 5.5, 40°C, native invertase INVB	Zymomonas mobilis
12149	-	sucrose	pH 5.0, 35°C, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5	-	assay at, invertase INVA	Zymomonas mobilis
5.5	-	assay at, invertase INVB	Zymomonas mobilis
5.5	-	invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, deglycosylated invertase INVA, native invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter, native invertase INVB and deglycosylated invertase INVB	Zymomonas mobilis
6	-	native invertase INVA	Zymomonas mobilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.517	-	sucrose	pH 5.0, 35°C, native invertase INVA	Zymomonas mobilis
107	-	sucrose	pH 5.0, 35°C, deglycosylated invertase INVA	Zymomonas mobilis
122	-	sucrose	pH 5.5, 40°C, native invertase INVB	Zymomonas mobilis
164	-	sucrose	pH 5.5, 40°C, deglycosylated invertase INVB	Zymomonas mobilis
208	-	sucrose	pH 5.5, 40°C, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis
296	-	sucrose	pH 5.0, 35°C, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter	Zymomonas mobilis