Home About us MoEF Contact us Sitemap Tamil Website  
About Envis
Whats New
Research on Microbes
Microbiology Experts
Online Submission
Access Statistics

Site Visitors

blog tracking

International Journal of Biological Macromolecules
Volume 164, 2020, 4318-4328

Stabilization of ?-transaminase from Pseudomonas fluorescens by immobilization techniques

SusanaVelasco-Lozanoa,b, Erienne Jacksonc, Magdalena Ripollc, FernandoLópez-Gallegoa,b,d, Lorena Betancorc

Catálisis Heterogénea en Síntesis Orgánicas Selectivas, Instituto de Síntesis Química y Catálisis Homogénea (ISQCH-CSIC), University of Zaragoza, Pedro Cerbuna, 12, 50009 Zaragoza, Spain.


Transaminases are a class of enzymes with promising applications for the preparation and resolution of a vast diversity of valued amines. Their poor operational stability has fueled many investigations on its stabilization due to their biotechnological relevance. In this work, we screened the stabilization of the tetrameric ω-transaminase from Pseudomonas fluorescens (PfωTA) through both carrier-bound and carrier-free immobilization techniques. The best heterogeneous biocatalyst was the PfωTA immobilized as cross-linked enzyme aggregates (PfωTA-CLEA) which resulted after studying different parameters as the precipitant, additives and glutaraldehyde concentrations. The best conditions for maximum recovered activity (29 %) and maximum thermostability at 60ºC and 70ºC (100 % and 71 % residual activity after 1 h, respectively) were achieved by enzyme precipitation with 90% acetone or ethanol, in presence of BSA (100 mg/mL) and employing glutaraldehyde (100 mM) as cross-linker. Studies on different conditions for PfωTA-CLEA preparation yielded a biocatalyst that exhibited 31 and 4.6 times enhanced thermal stability at 60°C and 70°C, respectively, compared to its soluble counterpart. The PfωTA-CLEA was successfully used in the bioamination of 4-hydroxybenzaldehyde to 4-hydroxybenzylamine. To the best of our knowledge, this is the first report describing a transaminase cross-linked enzyme aggregates as immobilization strategy to generate a biocatalyst with outstanding thermostability.

Keywords: PLP, Cross-linked enzyme aggregates, Biocatalysis.

Copyright © 2005 ENVIS Centre ! All rights reserved
This site is optimized for 1024 x 768 screen resolution