@article {5239,
	title = {Enzyme stabilization by domain insertion into a thermophilic protein.},
	journal = {Protein Engineering, Design \& Selection : PEDS},
	volume = {22},
	year = {2009},
	month = {2009 Oct},
	pages = {615-23},
	abstract = {<p>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{\textquoteright}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 degrees C without any compromise in EI activity. Our analysis suggests that the improved kinetic stability at 37 degrees 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.</p>
},
	keywords = {Aspergillus, Bacterial Proteins, Carrier Proteins, Enzyme Stability, Fungal Proteins, Glycoside Hydrolases, Kinetics, Models, Molecular, Protein Engineering, Protein Folding, Protein Structure, Pyrococcus, Recombinant Fusion Proteins, Temperature, Tertiary, Urea},
	issn = {1741-0134},
	doi = {10.1093/protein/gzp044},
	author = {Kim, Chung-Sei and Pierre, Brennal and Ostermeier, Marc and Looger, Loren L and Kim, Jin Ryoun}
}
