Research articles
ScienceAsia 50 (2024):ID 2024015 1-6 |doi:
10.2306/scienceasia1513-1874.2024.015
Chemical modification and blocking ratio assessment of Taq
DNA polymerases using a 2,4,6-trinitrobenzenesulfonic acid
(TNBS)-based assay
Jin Wanga, Wenhao Hua, Heqing Wanga, Hong Daia, Guochun Wanga, Zhidan Luoa,b,*, Henghao Xua,b,*
ABSTRACT: Mainstreaming modification approaches of commercial hot-start Taq DNA polymerases include antibody
modification, nucleic acid adaptor modification, and chemical modification, among which the chemical modification
possesses merits of lower cost and higher efficiency excluding animal-derived matters compared with the former
two. In chemical modification, the overall modification ratio of lysine residues is a key parameter to indicate the
blocking ratio of modified enzyme without compromising its stability. In this study, 2,4,6-trinitrobenzenesulfonic acid
(TNBS)-based assay was applied for chemically modified Taq DNA polymerase activity blocking ratio for the first time.
The results showed that at the modification ratio of about 50%, the enzyme activity was completely blocked. This
detection method?s features were easy-operation and time-saving for the screening of optimized anhydride modified
Taq DNA polymerase in multiple incubation conditions. Notably, this new protocol provided a practical reference for
identification of novel chemical compounds that can modify DNA polymerases, thereby facilitating the development of
advanced enzymes.
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| a |
Jiangsu Key Laboratory of Marine Biological Resources and Environment, Jiangsu Key Laboratory of Marine
Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005 China |
| b |
Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang 222005
China |
* Corresponding author, E-mail: lzd@jou.edu.cn, xuhh@jou.edu.cn
Received 10 Jan 2023, Accepted 20 May 2023
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