Liebherr, Raphaela B. and Renner, Max and Gorris, Hans H. (2014) A Single Molecule Perspective on the Functional Diversity of in Vitro Evolved beta-Glucuronidase. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 136 (16). pp. 5949-5955. ISSN 0002-7863,
Full text not available from this repository. (Request a copy)Abstract
The mechanisms that drive the evolution of new enzyme activity have been investigated by comparing the kinetics of wild-type and in vitro evolved beta-glucuronidase (GUS) at the single molecule level. Several hundred single GUS molecules were separated in large arrays of 62 500 ultrasmall reaction chambers etched into the surface of a fused silica slide to observe their individual substrate turnover rates in parallel by fluorescence microscopy. Individual GUS molecules feature long-lived but divergent activity states, and their mean activity is consistent with classic Michaelis-Menten kinetics. The large number of single molecule substrate turnover rates is representative of the activity distribution within an entire enzyme population. Partially evolved GUS displays a much broader activity distribution among individual enzyme molecules than wild-type GUS. The broader activity distribution indicates a functional division of work between individual molecules in a population of partially evolved enzymes that-as so-called generalists-are characterized by their promiscuous activity with many different substrates.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ENZYME MOLECULES; ESCHERICHIA-COLI; CATALYZED-HYDROLYSIS; DYNAMIC DISORDER; WILD-TYPE; PROTEIN; GALACTOSIDASE; EVOLUTION; THERMOSTABILITY; EVOLVABILITY; |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Nov 2019 09:17 |
| Last Modified: | 14 Nov 2019 09:17 |
| URI: | https://pred.uni-regensburg.de/id/eprint/10297 |
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