Structural highlights
Function
[SAV_STRAV] The biological function of streptavidin is not known. Forms a strong non-covalent specific complex with biotin (one molecule of biotin per subunit of streptavidin).
Publication Abstract from PubMed
Artificial metalloenzymes (ArMs hereafter) combine attractive features of both homogeneous catalysts and enzymes and offer the potential to implement new-to-nature reactions in living organisms. Herein we present an E. coli surface display platform for streptavidin (Sav hereafter) relying on an Lpp-OmpA anchor. The system was used for the high throughput screening of a bioorthogonal CpRu-based artificial deallylase (ADAse) that uncages an allylcarbamate-protected aminocoumarin 1. Two rounds of directed evolution afforded the double mutant S112M-K121A that displayed a 36-fold increase in surface activity vs. cellular background and a 5.7-fold increased in vitro activity compared to the wild type enzyme. The crystal structure of the best ADAse reveals the importance of mutation S112M to stabilize the cofactor conformation inside the protein.
E. coli surface display of streptavidin for directed evolution of an allylic deallylase.,Heinisch T, Schwizer F, Garabedian B, Csibra E, Jeschek M, Vallapurackal J, Pinheiro VB, Marliere P, Panke S, Ward TR Chem Sci. 2018 May 24;9(24):5383-5388. doi: 10.1039/c8sc00484f. eCollection 2018 , Jun 28. PMID:30079176[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Heinisch T, Schwizer F, Garabedian B, Csibra E, Jeschek M, Vallapurackal J, Pinheiro VB, Marliere P, Panke S, Ward TR. E. coli surface display of streptavidin for directed evolution of an allylic deallylase. Chem Sci. 2018 May 24;9(24):5383-5388. doi: 10.1039/c8sc00484f. eCollection 2018 , Jun 28. PMID:30079176 doi:http://dx.doi.org/10.1039/c8sc00484f
