TopicDirected enzyme evolution

presented by IFRAH ISHAQ

UNIVERSITY OF THE PUNJAB,LAHORE

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.

Steps of Directed enzyme evolution

1. Selection Of Gene2. A Library Of Mutant Genes3. Expression Of Mutant Genes4. Proteins Are Screened & Selected5. Variants With Modified Activity

Diagram representation

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

Goal: better luciferase

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.

Cephalosporins

Produced through intermediate

7-aminocephalosporanic acid

c

Atorvastatin drug• Atorvastatin• Is A Cholesterol Lowering Drug• Enzyme:2-deoxyribose- 5-phosphate

Aldolase (DERA)• Enzyme Target By Directed Enzyme

Evolution.

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.

Thank you!!!