Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21 (DE3), cloning in Escherichia coli DH5alpha, plasmid pET30 (Ek/LIC), 37 degrees Celsius, LB and 2TY medium | Zymomonas mobilis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | chimeric fusion enzyme INVA-CBD gene produced from enzyme gene invA and binding domain gene bcd from Cellulomonas fimi | Zymomonas mobilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
sucrose | - |
Zymomonas mobilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
51 | - |
sucrose | pH 5.5, 30°C, free chimeric fusion enzyme INVA-CBD | Zymomonas mobilis | |
110 | - |
sucrose | pH 5.5, 30°C, free INVA | Zymomonas mobilis | |
153 | - |
sucrose | pH 5.5, 30°C, cellulose immobilized chimeric fusion enzyme INVA-CBD | Zymomonas mobilis | |
189 | - |
sucrose | pH 5.5, 30°C, Nylon-6 immobilized INVA | Zymomonas mobilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
67000 | - |
6xHis-tagged enzyme INVA, 57 kDa from INVA, SDS-PAGE | Zymomonas mobilis |
78000 | - |
chimeric fusion enzyme INVA-CBD, 57 kDa from INVA, SDS-PAGE | Zymomonas mobilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Zymomonas mobilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
disrupted cells centrifuged, supernatant and pellet, combined purification and immobilization of chimeric gene product on Avicel (cellulose), or purification of 6-His-tagged enzyme by nickel affinity chromatography with Ni-NTA resin, elution with imidazol gradient (10-250 mM), dialyzed against 50 mM Tris-HCl, pH 7.0, and following immobilization on Nylon-6 microbeads | Zymomonas mobilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
20.6 | - |
cellulose immobilized chimeric fusion enzyme INVA-CBD | Zymomonas mobilis |
25.6 | - |
free chimeric fusion enzyme INVA-CBD | Zymomonas mobilis |
39 | - |
free INVA | Zymomonas mobilis |
456 | - |
Nylon-6 immobilized INVA | Zymomonas mobilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-D-glucopyranosyl-(1-2)-beta-D-fructofuranose | sucrose in 50 mM sodium acetate buffer, pH 5.5, 30 degrees Celsius | Zymomonas mobilis | glucose | - |
? | |
sucrose + H2O | - |
Zymomonas mobilis | beta-D-fructose + alpha-D-glucose | - |
? |
Synonyms | Comment | Organism |
---|---|---|
invertase | INVA and chimeric fusion enzyme INVA-CBD | Zymomonas mobilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
immobilized and free enzyme, INVA and chimeric fusion enzyme INVA-CBD, 50 mM acetate buffer, pH 5.5 | Zymomonas mobilis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
17 | 50 | INVA: same optimal temperature at 30°C, and similar course above that, retaining about 60% activity of the immobilized enzyme at 40°C, about 40% of the free enzyme, below 20% activity retained at 50°C | Zymomonas mobilis |
25 | 50 | chimeric fusion enzyme INVA-CBD, immobilized enzyme with higher activities above optimal temperature of 30°C, close to 60% activity at 40°C retained, while in free enzyme only about 20%, about 40% retained at 45-50°C in the immobilized enzyme while only about 20% in the free enzyme, at 25°C immobilized enzyme with about 40% activity, free enzyme with about 70% | Zymomonas mobilis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 40 | chimeric fusion enzyme INVA-CBD: free enzyme retains 90-100% activity at 25°C, while immobilized one drops to about 42% after 3 h, at 40°C free chimeric fusion enzyme INVA-CBD retains 20% activity after 20 min, while immobilized enzyme's activity ceases | Zymomonas mobilis |
30 | 50 | INVA: 100% activity retained after 7 h in free and immobilized INVA at 30°C, about 50% retained at 40°C in both enzyme forms, after 1 h at 50°C INVA activity ceases quickly retaining less than 20% activity while the immobilized INVA retains 70% activity after 10 min, 50 mM acetate buffer, pH 5.5 | Zymomonas mobilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
immobilized and free enzyme, INVA and chimeric fusion enzyme INVA-CBD, 30°C, pH 3.5-5.5: 50 mM acetate buffer, pH 6-7.5: 50 mM sodium citrate phosphate buffer, pH 7.5-8.5: 20 mM Tris-HCl buffer | Zymomonas mobilis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3.5 | 7.5 | chimeric fusion enzyme INVA-CBD: highest activity at pH 5.5, about 80% retained in immobilized enzyme at pH 6, in free enzyme about 60%. Both types with less than 50% activity below pH 5 and above pH 6.5 | Zymomonas mobilis |
3.5 | 8.5 | INVA: about 50% activity retained between pH 4.5 and 6.5 in the immobilized enzyme, about 40% in the free enzyme, no activity remains at pH 8.5 in both forms, none in the free enzyme at pH 3.5, about 30% in the immobilized enzyme | Zymomonas mobilis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
259 | - |
sucrose | Nylon-6 immobilized INVA | Zymomonas mobilis | |
283 | - |
sucrose | cellulose immobilized chimeric fusion enzyme INVA-CBD | Zymomonas mobilis | |
313 | - |
sucrose | free INVA | Zymomonas mobilis | |
393 | - |
sucrose | free chimeric fusion enzyme INVA-CBD | Zymomonas mobilis |