Enzyme stabilization by domain insertion into a thermophilic protein

22Citations
Citations of this article
48Readers
Mendeley users who have this article in their library.

This article is free to access.

Abstract

Insufficient kinetic stability of exoinulinase (EI) restricts its application in many areas including enzymatic transformation of inulin for production of ultra-high fructose syrup and oligofructan, as well as fermentation of inulin into bioethanol. The conventional method for enzyme stabilization involves mutagenesis and therefore risks alteration of an enzyme's desired properties, such as activity. Here, we report a novel method for stabilization of EI without any modification of its primary sequence. Our method employs domain insertion of an entire EI domain into a thermophilic scaffold protein. Insertion of EI into a loop of a thermophilic maltodextrin-binding protein from Pyrococcus furiosus (PfMBP) resulted in improvement of kinetic stability (the duration over which an enzyme remains active) at 37° C without any compromise in EI activity. Our analysis suggests that the improved kinetic stability at 37° C might originate from a raised kinetic barrier for irreversible conversion of unfolded intermediates to completely inactivated species, rather than an increased energy difference between the folded and unfolded forms. © The Author 2009. Published by Oxford University Press. All rights reserved.

Cite

CITATION STYLE

APA

Kim, C. S., Pierre, B., Ostermeier, M., Looger, L. L., & Kim, J. R. (2009). Enzyme stabilization by domain insertion into a thermophilic protein. Protein Engineering, Design and Selection, 22(10), 615–623. https://doi.org/10.1093/protein/gzp044

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free