Rational Engineering of Heparinase III from Pedobacter schmidteae to Improve the Thermostability via Distal Mutagenesis

Key Points

• The aim is to enhance the thermostability of Heparinase III from Pedobacter schmidteae through distal mutagenesis.
• Engineering of Heparinase III (M2a) from Pedobacter schmidteae via mutagenesis.
• Analysis of intramolecular interactions and protein surface charge using molecular dynamics simulations.
• Comparison of thermostability and specific activity of M2a relative to wild type (WT).
• Thermostability of M2a was increased by 23.66 °C compared to WT.
• Specific activity of M2a was 104.16% relative to WT.
• Enhanced local rigidity and increased surface potential contributed to stability improvement.