Literature summary for 3.2.1.26 extracted from

Vallejo-Becerra, V.; Vasquez-Bahena, J.M.; Santiago-Hernandez, J.A.; Hidalgo-Lara, M.E.
Immobilization of the recombinant invertase INVB from Zymomonas mobilis on Nylon-6 (2008), J. Ind. Microbiol. Biotechnol., 35, 1289-1295.

Search for more literature for 3.2.1.26

Cloned(Commentary)

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

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

Inhibitors	Comment	Organism	Structure
sucrose	-	Zymomonas mobilis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
51	-	sucrose	pH 5.5, 30Â°C, free chimeric fusion enzyme INVA-CBD	Zymomonas mobilis
110	-	sucrose	pH 5.5, 30Â°C, free INVA	Zymomonas mobilis
153	-	sucrose	pH 5.5, 30Â°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 [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

Organism	UniProt	Comment	Textmining
Zymomonas mobilis	-	-	-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms	Comment	Organism
invertase	INVA and chimeric fusion enzyme INVA-CBD	Zymomonas mobilis

Temperature Optimum [°C]

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 Range [°C]

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 [°C]

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

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 Range

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 [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

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Release 2023.1 (January 2023)