Disulfide Bond Engineering of an Endoglucanase from Penicillium verruculosum to Improve Its Thermostability
| Autor: | A. V. Bashirova, Alexander V. Gusakov, Pavel A. Volkov, Mehdi D. Davari, Arkady P. Sinitsyn, Subrata Pramanik, V. A. Nemashkalov, Ulrich Schwaneberg, Aleksandra M. Rozhkova, Ivan N. Zorov |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
cellulose biodegradation 030106 microbiology rational design disulfide bonds Cellulase Catalysis Inorganic Chemistry lcsh:Chemistry 03 medical and health sciences Physical and Theoretical Chemistry endoglucanase Molecular Biology Incubation lcsh:QH301-705.5 Spectroscopy Thermostability chemistry.chemical_classification cellulase biology Chemistry Organic Chemistry Rational design protein engineering General Medicine Protein engineering Combinatorial chemistry thermostability Computer Science Applications 030104 developmental biology Enzyme lcsh:Biology (General) lcsh:QD1-999 ddc:540 biology.protein Specific activity |
| Zdroj: | International Journal of Molecular Sciences, Vol 20, Iss 7, p 1602 (2019) International Journal of Molecular Sciences Volume 20 Issue 7 International journal of molecular sciences 20(7), 1602 (2019). doi:10.3390/ijms20071602 special issue: "Special Issue "Industrial Enzymes: Structure, Function and Applications" / Special Issue Editors: Prof. Dr. Dietmar Haltrich, Guest Editor; Prof. Roland Ludwig, Guest Editor" |
| DOI: | 10.18154/rwth-2019-05816 |
| Popis: | Endoglucanases (EGLs) are important components of multienzyme cocktails used in the production of a wide variety of fine and bulk chemicals from lignocellulosic feedstocks. However, a low thermostability and the loss of catalytic performance of EGLs at industrially required temperatures limit their commercial applications. A structure-based disulfide bond (DSB) engineering was carried out in order to improve the thermostability of EGLII from Penicillium verruculosum. Based on in silico prediction, two improved enzyme variants, S127C-A165C (DSB2) and Y171C-L201C (DSB3), were obtained. Both engineered enzymes displayed a 15&ndash 21% increase in specific activity against carboxymethylcellulose and &beta glucan compared to the wild-type EGLII (EGLII-wt). After incubation at 70 ° C for 2 h, they retained 52&ndash 58% of their activity, while EGLII-wt retained only 38% of its activity. At 80 ° C, the enzyme-engineered forms retained 15&ndash 22% of their activity after 2 h, whereas EGLII-wt was completely inactivated after the same incubation time. Molecular dynamics simulations revealed that the introduced DSB rigidified a global structure of DSB2 and DSB3 variants, thus enhancing their thermostability. In conclusion, this work provides an insight into DSB protein engineering as a potential rational design strategy that might be applicable for improving the stability of other enzymes for industrial applications. |
| Databáze: | OpenAIRE |
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