This document outlines the 9 key steps to produce and purify crude enzyme extracts from microbial sources which are: 1) isolating microbes, 2) screening microbes for enzyme production, 3) optimizing production medium and growing microbes, 4) disrupting cells to release intracellular enzymes, 5) separating solids from liquids, 6) concentrating the extract, 7) purifying the extract through techniques like dialysis, chromatography, and electrophoresis, 8) further processing the extract through crystallization, lyophilization or drying, and 9) obtaining the final purified enzyme product.

1. Production and purification of
crude enzyme extracts from plant,
animal and microbial sources
Ms. S. KARTHIKADEVI., M.Tech.,
Assistant Professor
Department of Biotechnology
Vivekanandha College of Engineering for Women
Namakkal (Dt).

3. Microbial sources
1. Isolation of microbes :

4. 2. Screening of microbes for enzyme production :
3. Enzyme production :

5. ➢ Medium optimization:

6. 4. Cell disruption :
➢ Intracellular Enzymes
➢Ex: Glucose isomerase, Dnase, Rnase etc…

9. 5. Solid - liquid separation:
6. Concentration: (Ammonium sulphate precipitation)

10. 7. Purification:
1. Dialysis:

11. 2. SDS PAGE:

13. 3. Western blotting:

15. 3. Chromatography purification:

16. Ion Exchange Chromatography

17. Affinity Chromatography

18. 8. Finishing:
➢ Crystallization :

19. ➢ Lyophilisation :

20. ➢ Drying :

21. 9. Final product:

22. THANK YOU