Vanella, Rosario and Nash, Michael A. and Fernández De Santaella, Jaime and Ren, Jin. (2023) Enzyme Cascade with Horseradish Peroxidase Readout for High-Throughput Screening and Engineering of Human Arginase-1. Analytical Chemistry, 95 (18). pp. 7150-7157.
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Official URL: https://edoc.unibas.ch/96381/
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Abstract
We report an enzyme cascade with horseradish peroxidase-based readout for screening human arginase-1 (hArg1) activity. We combined the four enzymes hArg1, ornithine decarboxylase, putrescine oxidase, and horseradish peroxidase in a reaction cascade that generated colorimetric or fluorescent signals in response to hArg1 activity and used this cascade to assay wild-type and variant hArg1 sequences as soluble enzymes and displayed on the surface of Escherichia coli. We screened a curated 13-member hArg1 library covering mutations that modified the electrostatic environment surrounding catalytic residues D128 and H141, and identified the R21E variant with a 13% enhanced catalytic turnover rate compared to wild type. Our scalable one-pot single-step arginase assay with continuous kinetic readout is amenable to high-throughput screening and directed evolution of arginase libraries and testing drug candidates for arginase inhibition.
Faculties and Departments: | 05 Faculty of Science > Departement Chemie > Chemie > Synthetic Systems (Nash) |
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UniBasel Contributors: | Vanella, Rosario and Nash, Michael |
Item Type: | Article |
Article Subtype: | Further Journal Contribution |
Publisher: | American Chemical Society |
ISSN: | 0003-2700 |
e-ISSN: | 1520-6882 |
Language: | English |
Language: | English |
Identification Number: | |
edoc DOI: | |
Last Modified: | 15 May 2024 09:22 |
Deposited On: | 15 May 2024 09:22 |
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