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Literature summary extracted from
- 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
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 3.2.1.26 | 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 |
| 3.2.1.26 | 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)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.2.1.26 | 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
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.2.1.26 | 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 |  |
| 3.2.1.26 | 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 | |
| 3.2.1.26 | Ni2+ | inhibits native invertase INVB and invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis | |
| 3.2.1.26 | 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 | |
| 3.2.1.26 | 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]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.26 | 41 | - |
sucrose | pH 5.0, 35°C, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis | |
| 3.2.1.26 | 50 | - |
sucrose | pH 5.5, 40°C, deglycosylated invertase INVB | Zymomonas mobilis | |
| 3.2.1.26 | 50 | - |
sucrose | pH 5.5, 40°C, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis | |
| 3.2.1.26 | 83 | - |
sucrose | pH 5.0, 35°C, deglycosylated invertase INVA | Zymomonas mobilis | |
| 3.2.1.26 | 98 | - |
sucrose | pH 5.5, 40°C, native invertase INVB | Zymomonas mobilis | |
| 3.2.1.26 | 110 | - |
sucrose | pH 5.0, 35°C, native invertase INVA | Zymomonas mobilis | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.2.1.26 | 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]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.26 | 46000 | - |
deglycosylated invertase INVB, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | 46000 | - |
native invertase INVB, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | 48000 | - |
invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | 57000 | - |
deglycosylated invertase INVA, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | 57000 | - |
native invertase INVA, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | 68000 | - |
invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE | Zymomonas mobilis |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.2.1.26 | Zymomonas mobilis | - |
- |
- |
| 3.2.1.26 | Zymomonas mobilis CDBB-B603 | - |
- |
- |
Posttranslational Modification
| EC Number | Posttranslational Modification | Comment | Organism |
|---|---|---|---|
| 3.2.1.26 | 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)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.2.1.26 | - |
Zymomonas mobilis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.2.1.26 | 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 | - |
? | |
| 3.2.1.26 | 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
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.2.1.26 | ? | x * 46000, deglycosylated invertase INVB, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | ? | x * 46000, native invertase INVB, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | ? | x * 48000, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | ? | x * 57000, deglycosylated invertase INVA, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | ? | x * 57000, native invertase INVA, SDS-PAGE | Zymomonas mobilis |
| 3.2.1.26 | ? | x * 68000, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter, SDS-PAGE | Zymomonas mobilis |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.2.1.26 | INVA | - |
Zymomonas mobilis |
| 3.2.1.26 | INVB | - |
Zymomonas mobilis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.26 | 30 | - |
native invertase INVA and deglycosylated invertase INVA | Zymomonas mobilis |
| 3.2.1.26 | 35 | - |
assay at, invertase INVA | Zymomonas mobilis |
| 3.2.1.26 | 35 | - |
invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis |
| 3.2.1.26 | 40 | - |
assay at, invertase INVB | Zymomonas mobilis |
| 3.2.1.26 | 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]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.26 | 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 |
| 3.2.1.26 | 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 |
| 3.2.1.26 | 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]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.26 | 56.92 | - |
sucrose | pH 5.0, 35°C, native invertase INVA | Zymomonas mobilis | |
| 3.2.1.26 | 8210 | - |
sucrose | pH 5.5, 40°C, deglycosylated invertase INVB | Zymomonas mobilis | |
| 3.2.1.26 | 8831 | - |
sucrose | pH 5.0, 35°C, deglycosylated invertase INVA | Zymomonas mobilis | |
| 3.2.1.26 | 10396 | - |
sucrose | pH 5.5, 40°C, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis | |
| 3.2.1.26 | 12000 | - |
sucrose | pH 5.5, 40°C, native invertase INVB | Zymomonas mobilis | |
| 3.2.1.26 | 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
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.26 | 5 | - |
assay at, invertase INVA | Zymomonas mobilis |
| 3.2.1.26 | 5.5 | - |
assay at, invertase INVB | Zymomonas mobilis |
| 3.2.1.26 | 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 |
| 3.2.1.26 | 6 | - |
native invertase INVA | Zymomonas mobilis |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.26 | 0.517 | - |
sucrose | pH 5.0, 35°C, native invertase INVA | Zymomonas mobilis | |
| 3.2.1.26 | 107 | - |
sucrose | pH 5.0, 35°C, deglycosylated invertase INVA | Zymomonas mobilis | |
| 3.2.1.26 | 122 | - |
sucrose | pH 5.5, 40°C, native invertase INVB | Zymomonas mobilis | |
| 3.2.1.26 | 164 | - |
sucrose | pH 5.5, 40°C, deglycosylated invertase INVB | Zymomonas mobilis | |
| 3.2.1.26 | 208 | - |
sucrose | pH 5.5, 40°C, invertase INVB expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis | |
| 3.2.1.26 | 296 | - |
sucrose | pH 5.0, 35°C, invertase INVA expressed in Pichia pastoris under the control of the strong AOX1 promoter | Zymomonas mobilis | |
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