1. Topic
Directed enzyme evolution
presented by
IFRAH ISHAQ

2. introduction
Directed Evolution is a general term used to describe
various molecular biology techniques that mimic
natural selection and are extremely useful for protein
engineering.
These techniques involve randomly introducing
mutations at the genetic level.

3. Steps of Directed enzyme evolution
1. Selection Of Gene
2. A Library Of Mutant Genes
3. Expression Of Mutant Genes
4. Proteins Are Screened & Selected
5. Variants With Modified Activity

4. Diagram representation

5. OCCURRENCE OF MUTAGENESIS
The formation of mutation in the gene of
interest by the following ways:
 Error Prone PCR
 DNA Shuffling
 Saturation Mutation
 Chemical Mutagenesis

6. Goal: better luciferase

7. Industrial Enzymes Activity Improved By Directed Enzyme Evolution
Some of the examples explained below:
 Cephalosporins: class of antibiotic
– produced via the intermediate 7-
aminocephalosporanic acid (7-ACA).
– Directed evolution has been used to improve the
activity of cephalosporin acylases to produce
these intermediates from adipyl-7-ACA or
cephalosporin C.

8. Cephalosporins
Produced through
intermediate
7-aminocephalosporanic acid
c

9. Atorvastatin drug
• Atorvastatin
• Is A Cholesterol Lowering Drug
• Enzyme:2-deoxyribose- 5-phosphate
Aldolase (DERA)
• Enzyme Target By Directed Enzyme
Evolution.

10. conclusion
 Directed evolution can be a powerful tool taking
advantage of nature’s power to improve upon itself
 Used in a wide variety of applications for protein
improvement – stability, activity, substrate
specificity, etc
 Potential for genetically engineering improved
drugs or crops
 Ultimately, combining tools will lead to better
understanding and applications.

11. Thank you!!!