Engineering ionic liquid-tolerant cellulases for biofuels production
| Autor: | Douglas S. Clark, Paul W. Wolski, Harvey W. Blanch, Craig M. Dana |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Ionic Liquids Lignocellulosic biomass Bioengineering Cellulase Protein Engineering 01 natural sciences Biochemistry Fungal Proteins 03 medical and health sciences chemistry.chemical_compound Hydrolysis Enzymatic hydrolysis Enzyme Stability Cellulases Cellulose Molecular Biology Trichoderma reesei Fungal protein Chromatography biology 010405 organic chemistry fungi Imidazoles Aminoacyltransferases biology.organism_classification 0104 chemical sciences 030104 developmental biology Talaromyces chemistry Biofuels Ionic liquid biology.protein Directed Molecular Evolution Biotechnology |
| Zdroj: | Protein Engineering Design and Selection. 29:117-122 |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/gzv066 |
| Popis: | Dissolution of lignocellulosic biomass in certain ionic liquids (ILs) can provide an effective pretreatment prior to enzymatic saccharification of cellulose for biofuels production. Toward the goal of combining pretreatment and enzymatic hydrolysis, we evolved enzyme variants of Talaromyces emersonii Cel7A to be more active and stable than wild-type T. emersonii Cel7A or Trichoderma reesei Cel7A in aqueous-IL solutions (up to 43% (w/w) 1,3-dimethylimdazolium dimethylphosphate and 20% (w/w) 1-ethyl-3-methylimidazolium acetate). In general, greater enzyme stability in buffer at elevated temperature corresponded to greater stability in aqueous-ILs. Post-translational modification of the N-terminal glutamine residue to pyroglutamate via glutaminyl cyclase enhanced the stability of T. emersonii Cel7A and variants. Differential scanning calorimetry revealed an increase in melting temperature of 1.9-3.9°C for the variant 1M10 over the wild-type T. emersonii Cel7A in aqueous buffer and in an IL-aqueous mixture. We observed this increase both with and without glutaminyl cyclase treatment of the enzymes. |
| Databáze: | OpenAIRE |
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