Expression, purification and immobilization of the intracellular invertase INVA, from Zymomonas mobilis on crystalline cellulose and Nylon-6.

Journal of Industrial Microbiology & Biotechnology
María de Los Angeles Calixto-RomoMaría Eugenia Hidalgo-Lara

Abstract

This paper presents two immobilization methods for the intracellular invertase (INVA), from Zymomonas mobilis. In the first method, a chimeric protein containing the invertase INVA, fused through its C-terminus to CBDCex from Cellulomonas fimi was expressed in Escherichia coli strain BL21 (DE3). INVA was purified and immobilized on crystalline cellulose (Avicel) by means of affinity, in a single step. No changes were detected in optimal pH and temperature when INVA-CBD was immobilized on Avicel, where values of 5.5 and 30 degrees C, respectively, were registered. The kinetic parameters of the INVA-CBD fusion protein were determined in both its free form and when immobilized on Avicel. Km and Vmax were affected with immobilization, since both showed an increase of up to threefold. Additionally, we found that subsequent to immobilization, the INVA-CBD fusion protein was 39% more susceptible to substrate inhibition than INVA-CBD in its free form. The second method of immobilization was achieved by the expression of a 6xHis-tagged invertase purified on Ni-NTA resin, which was then immobilized on Nylon-6 by covalent binding. An optimal pH of 5.5 and a temperature of 30 degrees C were maintained, subsequent to immobilization on Nylon...Continue Reading

References

Mar 1, 1991·Enzyme and Microbial Technology·J MansfeldH Dautzenberg
May 1, 1991·Agricultural and Biological Chemistry·H YanaseK Tonomura
Nov 1, 1996·Protein Science : a Publication of the Protein Society·M R BrayR A Warren
Aug 2, 2001·Bioresource Technology·S AhmadM Saleemuddin
Feb 2, 2008·Journal of Agricultural and Food Chemistry·Vanessa Vallejo-BecerraPedro Antonio García-Ruiz
Feb 1, 1987·Biotechnology and Bioengineering·C SimionescuS Dumitriu
Aug 21, 2008·Journal of Industrial Microbiology & Biotechnology·Vanessa Vallejo-BecerraMaría Eugenia Hidalgo-Lara

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Citations

Jul 16, 2010·Journal of Asian Natural Products Research·Wen-Li MeiHao-Fu Dai
Feb 26, 2010·Journal of Asian Natural Products Research·Wen-Jun DaiHao-Fu Dai
Feb 9, 2012·International Journal of Molecular Sciences·Hyemin ParkEun Gyo Lee
Sep 16, 2011·Enzyme Research·Abril Flores-MaltosCristóbal N Aguilar

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