Sun, Lianhong and Petrounia, Ioanna P. and Yagasaki, Makoto and Bandara, Geethani and Arnold, Frances H. (2001) Expression and stabilization of galactose oxidase in Escherichia coli by directed evolution. Protein Engineering, 14 (9). pp. 699-704. ISSN 0269-2139. doi:10.1093/protein/14.9.699. https://resolver.caltech.edu/CaltechAUTHORS:20111121-101225836
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Abstract
We have used directed evolution methods to express a fungal enzyme, galactose oxidase (GOase), in functional form in Escherichia coli. The evolved enzymes retain the activity and substrate specificity of the native fungal oxidase, but are more thermostable, are expressed at a much higher level (up to 10.8 mg/l of purified GOase), and have reduced negative charge compared to wild type, all properties which are expected to facilitate applications and further evolution of the enzyme. Spectroscopic characterization of the recombinant enzymes reveals a tyrosyl radical of comparable stability to the native GOase from Fusarium.
| Item Type: | Article | |||||||||
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| Additional Information: | © 2001 Oxford University Press. Received January 30, 2001. Accepted May 15, 2001. This work was supported by the Biotechnology Research and Development Corporation. | |||||||||
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| Subject Keywords: | directed evolution galactose oxidase random mutagenesis StEP recombination | |||||||||
| Issue or Number: | 9 | |||||||||
| DOI: | 10.1093/protein/14.9.699 | |||||||||
| Record Number: | CaltechAUTHORS:20111121-101225836 | |||||||||
| Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20111121-101225836 | |||||||||
| Official Citation: | Lianhong Sun, Ioanna P. Petrounia, Makoto Yagasaki, Geethani Bandara, and Frances H. Arnold Expression and stabilization of galactose oxidase in Escherichia coli by directed evolution Protein Eng. (2001) 14(9): 699-704 doi:10.1093/protein/14.9.699 | |||||||||
| Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | |||||||||
| ID Code: | 27885 | |||||||||
| Collection: | CaltechAUTHORS | |||||||||
| Deposited By: | Ruth Sustaita | |||||||||
| Deposited On: | 21 Nov 2011 19:07 | |||||||||
| Last Modified: | 09 Nov 2021 16:52 |
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