Immobilization of the recombinant invertase INVB from Zymomonas mobilis on Nylon-6.
The recombinant invertase INVB (re-INVB) from Zymomonas mobilis was immobilized on microbeads of Nylon-6, by means of covalent bonding. The enzyme was strongly and successfully bound to the support. The activity of the free and immobilized enzyme was determined, using 10% (w/v) sucrose, at a temperature ranging between 15 and 60 degrees C and a pH ranging between 3.5 and 7. The optimal pH and temperature for the immobilized enzyme were 5.5 and 25 degrees C, respectively. Immobilization of re-INVB on Nylon-6 showed no significant change in the optimal pH, but a difference in the optimal temperature was evident, as that for the free enzyme was shown to be 40 degrees C. The values for kinetic parameters were determined as: 984 and 98 mM for Kappm of immobilized and free re-INVB, respectively. Kappcat values for immobilized and free enzymes were 6.1x10(2) and 1.2x10(4) s(-1), respectively, and immobilized re-INVB showed Vappmax of 158.73 micromol h min(-1) mg(-1). Immobilization of re-INVB on Nylon-6 enhanced the thermostability of the enzyme by 50% at 30 degrees C and 70% at 40 degrees C, when compared to the free enzyme. The immobilization system reported here may have future biotechnological applications, owing to the simplicity of the immobilization technique, the strong binding of re-INVB to the support and the effective thermostability of the enzyme.
Vallejo-Becerra
Vanessa
V
Departamento de Biotecnología y Bioingeniería, CINVESTAV, IPN. Av. Instituto Politécnico Nacional 2508, Col. San Pedro Zacatenco, 07360, Mexico D. F., Mexico.
Vásquez-Bahena
Jazmín Magdalena
JM
Santiago-Hernández
José Alejandro
JA
Hidalgo-Lara
María Eugenia
ME
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002209:Caprolactam / Q000031:analogs & derivatives / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008567:Membranes, Artificial; D011108:Polymers / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017355:Zymomonas / Q000201:enzymology*; D043324:beta-Fructofuranosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D011108:Polymers; D011994:Recombinant Proteins; C009916:nylon 6; D002209:Caprolactam; D043324:beta-Fructofuranosidase