RESEARCH ARTICLE


A Novel Fluorogenic Coumarin Substrate for Monitoring Acid Phosphatase Activity at Low pH Environment



Desuo Yang1, Zongxiao Li1, Yubo Allan Diwu3, Hanzhuo Fu2, Jinfang Liao3, Chunmei Wei3, Zhenjun Diwu*, 3
1 Department of Chemistry & Chemical Engineering, Baoji University of Arts & Science, Shaanxi, China
2 Northwest University, Department of Chemistry, 229 North Taibei Road, Xian, Shaanxi, China
3 ABD Bioquest, Inc., 923 Thompson Place, Sunnyvale, CA 94085, USA


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© Yang et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the ABD Bioquest, Inc., 923 Thompson Place, Sunnyvale, CA 94085, USA; E-mail: jack@abdbioquest.com


Abstract

This article described the synthesis and application of 6-chloro-8-fluoro-4-methylumbelliferone phosphate (CF-MUP) in analyzing acid phosphatase activity. Compared to the existing MUP, the new coumarin phosphate, CF-MUP, demonstrateed much higher sensitivity and was more robust for detecting the activity of acid phosphatase than the classic substrate 4-methylumbelliferone phosphate (MUP). The product of enzyme reaction, 6-chloro-8-fluoro-4-methylumbelliferone (CF-MU) possesses strong fluorescence at ~450 nm with low pKa (4.7), high fluorescence quantum yield and pH independence in the physiological pH range. This new fluorescence dye, CF-MU, is a convenient tool for assays with buffer pH between 4.5 and 8.