Abstract
Controlling the catalytic properties of enzymes remain an important challenge in chemistry and biotechnology. We have recently established a strategy for altering enzyme specificity in which the addition of proxy monobodies, synthetic binding proteins, modulates the specificity of an otherwise unmodified enzyme. Here, in order to examine its broader applicability, we employed the strategy on Candida rugosa lipase 1 (CRL1), an enzyme with a tunnel-like substrate binding site. We successfully identified proxy monobodies that restricted the substrate specificity of CRL1 toward short-chain fatty acids. The successes with this enzyme system and a β-galactosidase used in the previous work suggest that our strategy can be applied to diverse enzymes with distinct architectures of substrate binding sites.
References
Mar 29, 1994·Biochemistry·P GrochulskiM Cygler
Jul 9, 1998·Protein Science : a Publication of the Protein Society·S BroccaR D Schmid
Nov 5, 1999·Annual Review of Microbiology·K E JaegerM T Reetz
Nov 21, 2000·Biotechnology and Bioengineering·N A TurnerE N Vulfson
Aug 30, 2002·Protein Engineering·Jutta SchmittJürgen Pleiss
Mar 1, 1965·The Biochemical Journal·N M GREEN
Jun 1, 2007·Journal of Agricultural and Food Chemistry·Li-Chiun LeeJei-Fu Shaw
Apr 2, 2010·Nature Structural & Molecular Biology·John WojcikShohei Koide
Sep 30, 2010·Journal of Agricultural and Food Chemistry·Chih-Chung YenJei-Fu Shaw
Apr 27, 2011·Proceedings of the National Academy of Sciences of the United States of America·Ryan N GilbrethShohei Koide
Dec 27, 2011·Journal of Molecular Biology·Akiko KoideShohei Koide
Aug 28, 2013·Proceedings of the National Academy of Sciences of the United States of America·Fern ShaOliver Hantschel
Oct 10, 2013·BioMed Research International·Neelam GurungVivek Rai
Jul 10, 2014·Current Opinion in Structural Biology·Wai Shun Mak, Justin B Siegel
Feb 5, 2015·Angewandte Chemie·Hans RenataFrances H Arnold
Feb 11, 2015·Proteins·Laura J Kingsley, Markus A Lill
Apr 17, 2015·The FEBS Journal·Kazuhiko IshikawaShotaro Yamaguchi
Jun 16, 2015·Industrial & Engineering Chemistry Research·Han XiaoHuimin Zhao
Sep 1, 2015·Nature Chemical Biology·Shun-Ichi TanakaShohei Koide