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Unformatted text preview: it.edu/OcwWeb/Biology/7-344Spring-2008/CourseHome/index.htm In vitro evolution of enzymes
• Enzymes evolved for functions inside a living organism, not for biotechnology
– Might need long-term stability – Might need activity in non-aqueous solutions • Produce new enzymes using recombinant DNA technology, but don’t know how by rational design • Use directed evolution Directed evolution experiment From Frances Arnold’s website: www.che.caltech.edu/groups/fha Some considerations…
• Don’t start with random sequences because there are too many (20N) • Instead start with lightly mutagenized gene (e.g., error-prone PCR) or high level of random mutations to small part of gene Most mutations are destabilizing, so simply increasing protein stability can increase mutational robustness Marginally stable parent protein stable
Stabilized parent protein unstable
These previously unacceptable mutati...
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