Molecular insights into particular mutations impact on Taq polymerase dynamics and structure: a molecular dynamics...

Area of Science:

• Biotechnology
• Enzyme Engineering
• Structural Biology

Background:

• Taq DNA polymerase (Taq pol) is crucial for biotechnology but requires enhanced thermostability.
• Current methods have not fully met the demand for more stable Taq pol variants.

Purpose of the Study:

• To engineer mutations in Taq pol to improve its structural stability.
• To investigate the impact of these mutations on enzyme dynamics and interactions.

Main Methods:

• Employed sequence-based and structure-based rational design for mutagenesis.
• Introduced five specific mutations into the Taq pol structure.
• Utilized molecular dynamics simulations to analyze mutant enzyme structures and dynamics.

Main Results:

• All mutant enzymes displayed more compact structures with increased internal hydrogen bonds and secondary structure content.
• Mutants TaqG389E and TaqS290K|A353K|L365E|N384E|G389E exhibited the least structural dynamics.
• Mutations generally enhanced non-bonded interactions and positively impacted overall structural stability.

Conclusions:

• The designed mutations successfully improved the structural stability of Taq pol.
• Mutants TaqG389E and TaqS290K|A353K|L365E|N384E|G389E are more stable than wild-type Taq pol due to reduced flexibility and enhanced interactions.