Total synthesis and biochemical characterization of mirror image barnase
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-b...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Royal Society of Chemistry, The
2017
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| Online Access: | http://hdl.handle.net/1721.1/109283 https://orcid.org/0000-0002-5508-0963 https://orcid.org/0000-0002-9383-2185 |
| Summary: | 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. |
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