Optimization of the masking molecule for active-site-protected immobilization of Taq DNA polymerase and its application

Gumjung Lim; Joungmok Kim; Jeong Hee Kim

doi:10.1016/j.ab.2012.09.033

Optimization of the masking molecule for active-site-protected immobilization of Taq DNA polymerase and its application

Anal Biochem. 2013 Jan 15;432(2):139-41. doi: 10.1016/j.ab.2012.09.033. Epub 2012 Oct 1.

Authors

Gumjung Lim¹, Joungmok Kim, Jeong Hee Kim

Affiliation

¹ Department of Chemistry, Kyung Hee University, Seoul 130-701, Republic of Korea.

PMID: 23036837
DOI: 10.1016/j.ab.2012.09.033

Abstract

The method of oriented and activity-preserved immobilization of biologically active proteins based on concepts of active-site masking and kinetic control was further developed in this study. Minimal requirements for the masking DNA molecule were found to be a 5'overhang of 5-7 nucleotides and a double-stranded region of 11-13 bp to retain approximately 70% of the enzyme activity. The amplification range of protected immobilized (PIM) Taq DNA polymerase was over 1.2 kb. These data suggest that PIM Taq DNA polymerase can be used for various commercial applications.

MeSH terms

Catalytic Domain
Enzymes, Immobilized / chemistry
Enzymes, Immobilized / metabolism
Kinetics
Polymerase Chain Reaction*
Reverse Transcriptase Polymerase Chain Reaction
Taq Polymerase / chemistry
Taq Polymerase / metabolism*

Substances

Enzymes, Immobilized
Taq Polymerase