ScienceAsia - Journal of The Science Society of Thailand

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.

Chemical modification and blocking ratio assessment of Taq DNA polymerases using a 2,4,6-trinitrobenzenesulfonic acid (TNBS)-based assay

Jin Wang^a, Wenhao Hu^a, Heqing Wang^a, Hong Dai^a, Guochun Wang^a, Zhidan Luo^a,b,, Henghao Xu^a,b,

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,*

Jin Wang^a, Wenhao Hu^a, Heqing Wang^a, Hong Dai^a, Guochun Wang^a, Zhidan Luo^a,b,, Henghao Xu^a,b,