Immobilization of S-methyltransferase

Area of Science:

• Biochemistry
• Enzyme Technology

Background:

• S-methyltransferase is crucial for various biological processes.
• Solubilizing and immobilizing enzymes are key steps in biocatalysis development.

Purpose of the Study:

• To solubilize and immobilize S-methyltransferase from pig liver microsomes.
• To evaluate the impact of immobilization on enzyme activity and stability.
• To investigate substrate specificity changes after immobilization.

Main Methods:

• Solubilization of S-methyltransferase using Zwittergent.
• Immobilization onto agarose and copolymerization with acrylamide.
• Assay of specific enzyme activity with varying substrate chain lengths (C1-C7).
• Assessment of enzyme stability at -20 degrees C.

Main Results:

• Agarose-bound S-methyltransferase exhibited a specific activity of 0.87 nmol/min/mg, while acrylamide-bound showed 0.55 nmol/min/mg.
• Soluble enzyme activity increased with substrate chain length (0.5 to 6.3 nmol/min/mg).
• Agarose immobilization resulted in constant specific activity (1.1 nmol/min/mg) across C1-C7 substrates.
• Agarose-immobilized enzyme showed twice the stability of the soluble form; acrylamide-immobilized enzyme was less stable.

Conclusions:

• Agarose immobilization is a promising method for enhancing S-methyltransferase stability and broadening substrate specificity.
• Immobilization on agarose normalizes substrate activity, making it suitable for diverse applications.
• Enzyme immobilization strategies significantly impact enzyme performance and stability.