Virus-Based Nanoreactors with GALT Activity for Classic Galactosemia Therapy

Area of Science:

• Biotechnology and Nanomedicine
• Enzyme Engineering
• Genetic Disorders Treatment

Background:

• Classic galactosemia is a metabolic disorder requiring enzyme-replacement therapy.
• Current therapies face challenges with enzyme stability and delivery.
• Virus-like nanoparticles offer a novel platform for enzyme encapsulation.

Purpose of the Study:

• To develop enzymatic nanoreactors by encapsulating galactose-1-phosphate uridylyl-transferase (GALT) into plant virus capsids.
• To evaluate the efficacy of these nanoreactors for potential enzyme-replacement therapy in classic galactosemia.
• To assess the encapsulation efficiency, catalytic activity, and cellular uptake of the GALT-loaded nanoreactors.

Main Methods:

• Molecular self-assembly strategy for GALT encapsulation into plant virus capsids.
• Determination of encapsulation efficiency and catalytic constants at varying GALT and coat protein ratios.
• Cellular internalization studies in fibroblast, kidney, and hepatocyte cell lines.

Main Results:

• Enzymatic nanoreactors were successfully produced with GALT encapsulated in virus capsids.
• Substrate affinity was slightly reduced compared to free GALT, with an activity rate of 16% of free GALT.
• Nanoreactors showed significant internalization into hepatocytes and other cell lines.

Conclusions:

• Virus-like nanoparticles containing GALT represent an innovative enzyme preparation.
• These enzymatic nanoreactors hold promise for the treatment of classic galactosemia.
• Further development could lead to an effective enzyme-replacement therapy.