title: Rational engineering of enzyme stability.
abstract: During the past 15 years there has been a continuous flow of reports describing proteins stabilized by the introduction of mutations. These reports span a period from pioneering rational design work on small enzymes such as T4 lysozyme and barnase to protein design, and directed evolution. Concomitantly, the purification and characterization of naturally occurring hyperstable proteins has added to our understanding of protein stability. Along the way, many strategies for rational protein stabilization have been proposed, some of which (e.g. entropic stabilization by introduction of prolines or disulfide bridges) have reasonable success rates. On the other hand, comparative studies and efforts in directed evolution have revealed that there are many mutational strategies that lead to high stability, some of which are not easy to define and rationalize. Recent developments in the field include increasing awareness of the importance of the protein surface for stability, as well as the notion that normally a very limited number of mutations can yield a large increase in stability. Another development concerns the notion that there is a fundamental difference between the "laboratory stability" of small pure proteins that unfold reversibly and completely at high temperatures and "industrial stability", which is usually governed by partial unfolding processes followed by some kind of irreversible inactivation process (e.g. aggregation). Provided that one has sufficient knowledge of the mechanism of thermal inactivation, successful and efficient rational stabilization of enzymes can be achieved.
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, PO Box 5040, N-1432 As. vincent.eijsink@ikbm.nlh.no
authors.forename: Vincent G H
authors.initials: VG
authors.lastname: Eijsink
authors.forename: Alexandra
authors.initials: A
authors.lastname: Bjørk
authors.forename: Sigrid
authors.initials: S
authors.lastname: Gåseidnes
authors.forename: Reidun
authors.initials: R
authors.lastname: Sirevåg
authors.forename: Bjørnar
authors.initials: B
authors.lastname: Synstad
authors.forename: Bertus
authors.initials: B
authors.lastname: van den Burg
authors.forename: Gert
authors.initials: G
authors.lastname: Vriend
mesh_terms: D001709:Biotechnology / Q000379:methods*; D004798:Enzymes / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*
chemical_list: D004798:Enzymes