Coupling S-adenosylmethionine-dependent methylation to growth: Design and uses

Area of Science:

• Biochemistry
• Molecular Biology
• Enzyme Engineering

Background:

• Methylation is crucial for various cellular processes.
• Improving enzyme activity is key for biotechnology and drug development.
• Existing methods for enzyme engineering have limitations.

Purpose of the Study:

• To develop a novel selection design coupling S-adenosylmethionine-dependent methylation to growth.
• To enhance the activity of methyltransferases and other enzymes.
• To apply this design for drug discovery and enzyme engineering in different organisms.

Main Methods:

• Coupling S-adenosylmethionine-dependent methylation to a growth phenotype.
• Employing adaptive laboratory evolution in Escherichia coli.
• Utilizing catechol O-methyltransferase and its inhibitors for drug discovery applications.
• Implementing the selection design in Saccharomyces cerevisiae.

Main Results:

• Achieved a 2-fold improvement in enzyme activities for N-type and O-type methyltransferases.
• Enhanced the activity of an acetyltransferase by linking it to a methylation pathway.
• Demonstrated successful application in drug discovery targeting catechol O-methyltransferase.
• Successfully implemented the selection design in both E. coli and S. cerevisiae.

Conclusions:

• The developed selection design effectively couples methylation to growth, enabling enzyme activity enhancement.
• This approach is versatile, applicable to various enzymes and organisms, including for drug discovery.
• The study provides a powerful tool for enzyme engineering and the development of novel therapeutics.