Issue 5, 2015

Total synthesis and biochemical characterization of mirror image barnase

Abstract

In this study we synthesized and characterized mirror image barnase (B. amyloliquefaciens ribonuclease). D-Barnase was identical to L-barnase, when analyzed by liquid chromatography and mass-spectrometry. Proteolysis of the mirror image enzyme revealed that in contrast to its native counterpart, D-barnase was completely stable to digestive proteases. In enzymatic assays, D-barnase had the reciprocal chiral specificity and was fully active towards mirror image substrates. Interestingly, D-barnase also hydrolyzed the substrate of the native chirality, albeit 4000 times less efficiently. This effect was further confirmed by digesting a native 112-mer RNA with the enzyme. Additional studies revealed that barnase accommodates a range of substrates with various chiralities, but the prime requirement for guanosine remains. These studies point toward using mirror image enzymes as modern agents in biotechnology.

Graphical abstract: Total synthesis and biochemical characterization of mirror image barnase

Supplementary files

Article information

Article type
Edge Article
Submitted
14 Dec 2014
Accepted
25 Feb 2015
First published
23 Mar 2015
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2015,6, 2997-3002

Author version available

Total synthesis and biochemical characterization of mirror image barnase

A. A. Vinogradov, E. D. Evans and B. L. Pentelute, Chem. Sci., 2015, 6, 2997 DOI: 10.1039/C4SC03877K

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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