Exploration of Enzyme Diversity by Integrating Bioinformatics with Expression Analysis and Biochemical Characterization
| Autor: | Zbynek Prokop, Petra Babková, Pavel Vanacek, Jan Brezovsky, Jiri Damborsky, Veronika Stepankova, Lukas Daniel, Eva Sebestova, Pavel Dvorak, Radka Chaloupková, Šárka Bidmanová |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification In silico 030106 microbiology General Chemistry Computational biology Biology Catalysis 03 medical and health sciences Structural bioinformatics 030104 developmental biology Enzyme chemistry Identification (biology) Sequence space (evolution) Gene Dehalogenase Haloalkane dehalogenase |
| Zdroj: | ACS Catalysis |
| ISSN: | 2155-5435 |
| DOI: | 10.1021/acscatal.7b03523 |
| Popis: | Millions of protein sequences are being discovered at an incredible pace, representing an inexhaustible source of biocatalysts. Here, we describe an integrated system for automated in silico screening and systematic characterization of diverse family members. The workflow consists of (i) identification and computational characterization of relevant genes by sequence/structural bioinformatics, (ii) expression analysis and activity screening of selected proteins, and (iii) complete biochemical/biophysical characterization and was validated against the haloalkane dehalogenase family. The sequence-based search identified 658 potential dehalogenases. The subsequent structural bioinformatics prioritized and selected 20 candidates for exploration of protein functional diversity. Out of these 20, the expression analysis and the robotic screening of enzymatic activity provided 8 soluble proteins with dehalogenase activity. The enzymes discovered originated from genetically unrelated Bacteria, Eukaryota, and also A... |
| Databáze: | OpenAIRE |
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