Study of alkaline phosphatase interaction with putrescine using multi-spectroscopic and docking methods

Area of Science:

• Biochemistry
• Enzyme kinetics
• Protein structure analysis

Background:

• Enzyme optimization is crucial for industrial applications.
• Solvent engineering, including polyamine use, stabilizes enzymes.
• Putrescine is a polyamine with potential enzyme-stabilizing properties.

Purpose of the Study:

• To investigate the effect of putrescine on bovine alkaline phosphatase (BALP) structure and activity.
• To understand the interaction mechanism between putrescine and BALP.
• To explore putrescine as a potential enzyme stabilizer.

Main Methods:

• UV-vis spectroscopy to study enzyme-ligand binding.
• Fluorescence spectroscopy to analyze enzyme conformational changes.
• Circular dichroism (CD) spectroscopy for structural analysis.
• Enzyme kinetics assays to determine activity changes.
• Molecular docking to predict binding interactions.

Main Results:

• Putrescine binds to BALP, altering its UV-vis and fluorescence spectra via static quenching.
• Putrescine interacts with BALP through van der Waals forces and hydrogen bonding.
• CD spectra indicate structural modifications in BALP upon putrescine binding.
• Putrescine activates BALP, as evidenced by kinetic parameter changes.
• Docking studies confirm hydrogen bonds, van der Waals forces, and hydrophobic interactions in the BALP-putrescine complex.

Conclusions:

• Putrescine effectively binds to and modifies the structure of bovine alkaline phosphatase.
• Putrescine enhances BALP activity, demonstrating its potential as an enzyme stabilizer.
• The interaction involves multiple non-covalent forces, elucidated by spectroscopic and computational methods.