Immobilization of the recombinant invertase INVB from Zymomonas mobilis on Nylon-6

Abstract

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.

Potential Production of Ethanol by Saccharomyces cerevisiae Immobilized and Coimmobilized with Zymomonas mobilis: Alternative for the Reuse of a Waste Organic

“…Other studies have been developed with immobilized algal cells to remove nutrients (N and P) from wastewater, phenol and hexavalent chromium [14][15][16][17]. Similarly, the immobilization of Zymomonas and Saccharomyces have been used for the bio-ethanol production from waste materials [7][8][9][10][18][19][20].…”

Stabilization of invertase by molecular engineering

“…Previously, enzymes have been frequently stabilized by crosslinking to solid supports and entrapment within matrices (Betancor and Luckarift, 2008;Bryjak et al, 2008;Dizge et al, 2008;Mateo, et al, 2007;Polizzi et al, 2007;Vallejo-Becerra et al, 2008). The relevant immobilization methods have been reviewed by Sheldon (2007).…”

Surface Modifications for Controlled and Optimized Cell Immobilization by Adsorption: Applications in Fibrous Bed Bioreactors Containing Recombinant Cells

“…Therefore, GA may react readily with the protein in the lipid bilayer of cell membrane [27,28]. Its aldehyde groups can also cross-link the amino groups on PEI-coated materials and the surfaces to promote cell-support interaction by chemical bonding [28,29].…”

“…Thus, it can be concluded that PEI-GA modification can be an affective method to improve cell immobilization on fibrous matrices [28,29]. PEI is non-toxic and has been approved by the FDA for use in potable water [22].…”

“…GA is well known for its wide application in biological cross-linking. Therefore, PEI-GA modified fibrous matrices should have a good potential for large-scale use in recombinant cell immobilization [28,29]. Note: Cells were harvested from the stationary phase and suspended in PBS buffer.…”