Modification of chimeric (2S, 3S)-butanediol dehydrogenase based on structural information
| Autor: | Kaito Mochizuki, Sadaharu Ui, Takuji Oyama, Tomohito Shimegi, Masami Kusunoki, Takashi Ohtsuki |
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| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular Dehydrogenase Biology Corynebacterium medicine.disease_cause Crystallography X-Ray Biochemistry Substrate Specificity Bacterial Proteins Structural Biology Catalytic Domain Klebsiella Enzyme Stability medicine Cloning Molecular chemistry.chemical_classification Mutation Brevibacterium saccharolyticum General Medicine Alcohol Oxidoreductases Enzyme chemistry Structural Homology Protein Butanediol dehydrogenase Mutagenesis Site-Directed Substrate specificity |
| Zdroj: | Protein and peptide letters. 22(3) |
| ISSN: | 1875-5305 |
| Popis: | A chimeric (2S, 3S)-butanediol dehydrogenase (cLBDH) was engineered to have the strict (S)-configuration specificity of the (2S, 3S)-BDH (BsLBDH) derived from Brevibacterium saccharolyticum as well as the enzymatic stability of the (2R, 3S)-BDH (KpMBDH) from Klebsiella pneumonia by swapping the domains of two native BDHs. However, while cLBDH possesses the stability, it lacks the specificity. In order to assist in the design a BDH having strict substrate specificity, an X-ray structural analysis of a cLBDH crystal was conducted at 1.58 A. The results obtained show some readily apparent differences around the active sites of cLBDH and BsLBDH. Based on this structural information, a novel (2S, 3S)-BDH having a preferred specificity was developed by introducing a V254L mutation into cLBDH. The influence of this mutation on the stability of cLBDH was not evaluated. Nevertheless, the technique described herein is an effective method for the production of a tailor-made BDH. |
| Databáze: | OpenAIRE |
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