Engineered disulfide bond

As for aqualysin I, the disulfide bond seems to be one of the causes of its thermostability, because the enzyme is unstable in the presence of 2-mercaptoethanol.15) Accordingly, increasing the stability of subtilisin E was attempted by engineering disulfide bonds at the sites corresponding to those of aqualysin I. 

Various biophysical probes indicate that CcP has two sites of interaction with cytochrome c although it is less clear if both sites are involved in electron transfer. To probe this question, CcP and cytochrome c have been site-specifically cross-linked using engineered disulfide bonds to generate a covalent complex that mimics the noncovalent complex. CcP in the covalent complex is unable to accept electrons from free ferrocytochrome c. A second cross-link was designed to block... 

Knight, E., Jr. Fathey, D. Human fibroblast interferon an improved purification. J. Biol. Chem. 1981,256, 3609-3611. Wells, J.A. Powers, D.B. In vivo formation and stability of engineered disulfide bonds in subtilisin. J. Biol. Chem. 1986, 261, 6564-6570. 

PeUequer, J.-L., Chen, S.-w. W. (2006). Multi-template approach to modeling engineered disulfide bonds. Proteins Structure, Function, and Bioinformatics, 65(1), 192—202. http //dx. doi.org/10.1002/prot.21059. 

D. Engineering Disulfide Bonds in Bamase VII. Chaperonin-Facilitated Protein Folding... 

Mansfeld J, Vriend G, Dijkstra BW, Veltman OR, van Den BB, Venema G, Ulbrich-Hofmann R, Eijsink VG (1997) Extreme stabilization of a thermolysin-like protease by an engineered disulfide bond. J Biol Chem 272 11152-11156 Matsuda A, Matsuyama K, Yamamoto K, Ichikawa S, Komatsu K (1987) Cloning and characterization of the genes for two distinct cephalosporin acylases from a Pseudomonas strain. J Bacteriol 169 5815-5820 Matsumura I, Wallingford JB, Surana NK, Vize PD, Ellington AD (1999) Directed evolution of the surface chemistry of the reporter enzyme beta-glucuronidase. Nat Biotechnol 17 696-701... 

Durrschmidt, P. Differentiation between conformational and autoproteolytic stability of the neutral protease from Bacillus stearothermophilus containing an engineered disulfide bond. EurJ Biochem 2001,268, 3612-3618. 

---