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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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Evolution of an Enzyme from a Noncatalytic Nucleic Acid Sequence
Rachel Gysbers1, Kha Tram2, Jimmy Gu3
11] Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main St. W., Hamilton, ON L8S 4K1, Canada [2] Origins Institute, McMaster University, 1280 Main St. W., Hamilton, ON L8S 4K1, Canada.
Scientific Reports
|June 20, 2015
Summary
Enzymes can emerge from functional polymers through molecular evolution. This study demonstrates DNA evolving into a catalytic DNAzyme, revealing key mutations driving this transformation.
Area of Science:
- Biochemistry
- Molecular Biology
- Origins of Life
Background:
- The origin of enzymes, crucial biological catalysts, from non-living matter is a long-standing scientific enigma.
- Understanding enzyme genesis is key to comprehending early life evolution and synthetic biology.
Purpose of the Study:
- To investigate the potential for functional polymers to evolve catalytic activity under laboratory conditions.
- To demonstrate that enzymes can arise from non-catalytic sequences through directed molecular evolution.
Main Methods:
- Selection and amplification of a 50-nucleotide DNA sequence encoding cattle albumin mRNA.
- Application of test-tube evolution to generate catalytic DNA (DNAzyme) with RNA-cleavage activity.
- Deep sequencing to identify mutations driving the evolution of catalytic function.
Main Results:
- A DNAzyme with significant RNA-cleavage catalytic activity was successfully derived within weeks.
- Sequence analysis pinpointed seven critical nucleotides responsible for the emergence of enzymatic function.
- Deep sequencing revealed that specific mutations progressively enhanced molecular evolution and catalytic efficiency.
Conclusions:
- Enzymes can spontaneously arise from functional polymer sequences via permissive molecular evolution.
- This mechanism provides a plausible pathway for the natural origin of the vast array of enzymes found in biology.
- The findings support the potential for engineering novel catalytic molecules through directed evolution.
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