PEG modification increases thermostability and inhibitor resistance of Bst DNA polymerase.
Biosci Biotechnol Biochem
; 88(7): 768-775, 2024 Jun 21.
Article
in En
| MEDLINE
| ID: mdl-38734885
ABSTRACT
Polyethylene glycol modification (PEGylation) is a widely used strategy to improve the physicochemical properties of various macromolecules, especially protein drugs. However, its application in enhancing the performance of enzymes for molecular biology remains underexplored. This study explored the PEGylation of Bst DNA polymerase, determining optimal modification reaction conditions. In comparison to the unmodified wild-type counterpart, the modified Bst DNA polymerase exhibited significantly improved activity, thermal stability, and inhibitor tolerance during loop-mediated isothermal amplification. When applied for the detection of Salmonella in crude samples, the modified enzyme demonstrated a notably accelerated reaction rate. Therefore, PEGylation emerges as a viable strategy for refining DNA polymerases, helping in the development of novel molecular diagnostic reagents.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
Polyethylene Glycols
/
Enzyme Stability
/
DNA-Directed DNA Polymerase
Language:
En
Journal:
Biosci Biotechnol Biochem
Journal subject:
BIOQUIMICA
/
BIOTECNOLOGIA
Year:
2024
Type:
Article
Affiliation country:
China