Microbial Insulin Production powerpoint presentation

The Microbial Insulin Production presentation provides an in-depth examination of recombinant insulin production and the challenges associated with meeting the rising demand for this critical biopharmaceutical. The introduction highlights the surging number of diabetes patients, straining the production capacity and driving up costs. Recombinant human insulin is primarily manufactured using E. coli and Saccharomyces cerevisiae, although other expression systems like Pichia pastoris and transgenic plants are also explored.

The history section presumably discusses the development of insulin production, which has been a significant milestone in the treatment of diabetes. The document then examines the structure and function of insulin, a vital hormone for regulating blood sugar levels, and its synthesis within the body.

The core of the presentation focuses on the production of insulin through Recombinant DNA technology, employing a two-chain method. The bio-production steps are detailed, beginning with gene isolation and plasmid insertion, followed by transfection, medium preparation, bioreactor fermentation, crude product isolation, purification, and chain joining.

Various expression systems for insulin production are compared, including E. coli, which was the first system used in 1978, yeast (Saccharomyces cerevisiae and Pichia pastoris), and transgenic plants like Arabidopsis thaliana, tobacco, lettuce, and strawberry. Each system has its advantages and disadvantages, with yeast being noted for its ability to perform post-translational modifications (PTMs), although it can lead to immune responses in humans.

In conclusion, the document underscores the urgent need for more efficient insulin production methods due to the escalating diabetes rates and the limitations of current technologies. It emphasizes the necessity of innovative solutions to address the production capacity and cost issues to ensure a steady supply of insulin for the growing number of diabetes patients.

The references section cites several studies and articles that have contributed to the understanding and advancing of insulin production, including works by Baeshen et al., Alyas et al., and Riggs. These sources provide further insights into the scientific and technical aspects of insulin manufacturing and its historical context.
Source:
https://sciencecodons.com/2742-microbial-insulin-production/

Transcript

2. contents • Introduction • History • Structure, function and synthesis of insulin • Insulin production by Recombinant DNA technology: Two-chain method • Bio-production Steps • Different Expression Systems of insulin • Conclusion • References 2
3. Introduction rapid increase in the number of diabetic patients limitation in production capacity high production cost the demand for recombinant insulin Recombinant human insulin has been produced predominantly using E. coli and Saccharomyces cerevisiae for therapeutic use in human. 3
4. History 4
5. Biopharmaceutical products • Therapeutic monoclonal antibodies • the hormones • growth factors 5
6. Structure and function of insulin 6
7. Insulin synthesis 7
8. Insulin production by Recombinant DNA technology: Two-chain method 8
9. Bio-production Steps 1. Gene Isolation 2. Insertion into the Plasmid 9
10. Bio-production Steps 3. Transfection 4. Medium Preparation 5. Bioreactor Fermentation 10
11. Bio-production Steps 6. Crude Product Isolation 7. Purification 9. Chain Joining 11
12. Different Expression Systems of insulin • E. coli • Yeast: • Saccharomyces cerevisiae • Pichia pastoris • Transgenic Plants: • Arabidopsis thaliana • Tobacco and Lettuce Plants • Strawberry Plant 12
13. E. coli • the first expression system used to produce human insulin in 1978 • Advantages • Disadvantages • new technologies to solve problems 13
14. Yeast • another commercially-used expression system for heterogeneous protein production with recombinant technology because of its PTMs. • Advantages • Disadvantage: proteins synthesized in yeast have high-mannose N-glycosylation activates an immune response in humans shortens the half-life of recombinant proteins in vivo 14
15. Yeast: Saccharomyces cerevisiae • Produce: • Proinsulin • Fast-acting insulin • Long-acting insulin 15
16. Yeast: Pichia pastoris • presence of the methanol-inducible alcohol oxidase-1 promoter (AOX-1) • Heterogeneous protein expression attained by P. pastoris is approximately 30% of its total cell protein 16
17. Transgenic Plants • Plants do not have human pathogens and because they are eukaryotic, their PTM machinery is more similar to humans than that previously described in yeast. • Arabidopsis thaliana • Tobacco and Lettuce Plants • Strawberry Plant 17
18. Conclusion Dietary and lifestyle changes are causing dramatic increase in diabetes incidence all over the world. Both Type I and Type II diabetic patients use insulin, however late stage Type II diabetes patients require large doses of insulin as they develop insulin resistance. Current manufacturing technologies will not be able to meet the growing demand of insulin due to limitation in production capacity and high production cost. 18
19. References Baeshen, N. A., Baeshen, M. N., Sheikh, A., Bora, R. S., Ahmed, M. M. M., Ramadan, H. A., ... & Redwan, E. M. (2014). Cell factories for insulin production. Microbial cell factories, 13(1), 1-9. Alyas, J., Rafiq, A., Amir, H., Khan, S. U., Sultana, T., Ali, A., ... & Ahmad, A. (2021). Human Insulin: History, Recent Advances, and Expression Systems for Mass Production. Biomedical Research and Therapy, 8(9), 4540-4561. Riggs, A. D. (2021). Making, Cloning, and the Expression of Human Insulin Genes in Bacteria: The Path to Humulin. Endocrine Reviews, 42(3), 374-380. 19