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Role of some fermentation parameters on cyclosporin A production by a new isolate of Aspergillus terreus. Presented by SEEMA RENU.

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Presentation on theme: "Role of some fermentation parameters on cyclosporin A production by a new isolate of Aspergillus terreus. Presented by SEEMA RENU."— Presentation transcript:

1 Role of some fermentation parameters on cyclosporin A production by a new isolate of Aspergillus terreus. Presented by SEEMA RENU

2 Struture of Cyclosporin A Cyclosporin A is a cyclopeptide composed of 11 amino acids Cyclosporin A is a cyclopeptide composed of 11 amino acids It was first derived from submerged culture of aerobic fungi identified as strains of Trichoderma polysporum. It was first derived from submerged culture of aerobic fungi identified as strains of Trichoderma polysporum. Trichoderma polysporum currently identified as Tolypocladium inflatum also known as Tolypocladium niveum. Trichoderma polysporum currently identified as Tolypocladium inflatum also known as Tolypocladium niveum. T. inflatum also produces numerous other cyclosporins all composed of 11 amino acids differing from one another in only one amino acid. T. inflatum also produces numerous other cyclosporins all composed of 11 amino acids differing from one another in only one amino acid. For example the second amino acid in cyclosporine A is alpha amino butyric acid whereas that in cyclosporine B is alanine. For example the second amino acid in cyclosporine A is alpha amino butyric acid whereas that in cyclosporine B is alanine.

3 Discovery of Cyclosporin A It was discovered in the early 1970 by research workers at Sandoz, Ltd in Basel, Switzerland during a search for fungal metabolites with antibiotic properties. It was discovered in the early 1970 by research workers at Sandoz, Ltd in Basel, Switzerland during a search for fungal metabolites with antibiotic properties. One of the metabolites released, known as 24-556 had antifungal properties and very low toxicity toward the mammalian cells when tested at Sandoz. One of the metabolites released, known as 24-556 had antifungal properties and very low toxicity toward the mammalian cells when tested at Sandoz. The two components of 24-556 were termed as Cyclosporin A and Cyclosporin B where as Cycloporin A was much more active than Cyclosporine B. The two components of 24-556 were termed as Cyclosporin A and Cyclosporin B where as Cycloporin A was much more active than Cyclosporine B. Human transplantation was first performed in 1978 using cyclosporin A. Since then it has a major role in allogenic transplantation and autoimmune diseases. Human transplantation was first performed in 1978 using cyclosporin A. Since then it has a major role in allogenic transplantation and autoimmune diseases.

4 Mechanism of action Cyclosporin A works by selectively affecting the production of T and B lymphocytes produced in the bone marrow and thymus. Cyclosporin A works by selectively affecting the production of T and B lymphocytes produced in the bone marrow and thymus. Individuals with autoimmune diseases such as rheumatoid arthritis produces excessive amount of toxic molecules such as reactive oxygen intermediates that cause inflammation in the joints. Individuals with autoimmune diseases such as rheumatoid arthritis produces excessive amount of toxic molecules such as reactive oxygen intermediates that cause inflammation in the joints. Cy A reduces the inflammation by selectively acting against the production of these lymphocytes. Cy A reduces the inflammation by selectively acting against the production of these lymphocytes.

5 A macrophage engulfing a bacterium and releasing packets of toxic molecules (reactive oxygen intermediates) that breaks down and destroy the bacterium

6 Cyclosporin A and kidney damage The widespread use of cyclosporin A showed that it causes damage to the kidneys. The widespread use of cyclosporin A showed that it causes damage to the kidneys. Studies proved that the damage can be avoided by using cyclosporin A in lower amounts and in conjunction with a steroid called prednisone. Studies proved that the damage can be avoided by using cyclosporin A in lower amounts and in conjunction with a steroid called prednisone.

7 Fermentation parameters and Cy A production by A. Terreus The conditions affecting the production of Cy A by A. Terreus included the composition of the cultivation medium, fermentation time course, inoculum nature, medium volume, agitation rate and pH value. The conditions affecting the production of Cy A by A. Terreus included the composition of the cultivation medium, fermentation time course, inoculum nature, medium volume, agitation rate and pH value. Different fungi were investigated for Cy A production. Different fungi were investigated for Cy A production. – sign indicated no production of Cy A, ++ showed a moderate amount of Cy A production, whereas three +++ means higher production of Cy A as judged by the darkness of the colony color. – sign indicated no production of Cy A, ++ showed a moderate amount of Cy A production, whereas three +++ means higher production of Cy A as judged by the darkness of the colony color.

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9 The yield of the Cy A increased as the fermentation time course increased. The highest yield of Cy A in mg/ml was on the day 10. The biomass yield of A. terreus (g/l) was kept on decreasing after day 10.

10 The best yield of Cy A was obtained by a 48-h-old culture. As the age increased production of Cy A start to decrease.

11 Maximum yield of Cy A was obtained when the volume of the inoculum size was 2ml/100ml.

12 The size of the medium also affected the production of Cy A. A 250 ml Erlenmeyer flask containing production medium equal to 100ml afforded the highest production of Cy A

13 The formation of Cy A by A.terreus was also affected by the agitation rate. The maximum production was at 200 rpm

14 The production was maximum in an acidic medium with a pH 5.3. Cy A production decreased with an increase in pH.

15 Buffer is a partially neutralised acid that resists changes in pH. The results showed that on using citrate buffer the maximum yield (84.22 mg/L) of Cy A was obtained. Increased pH also affected the texture of the fungal pellets by making it weaker.

16 RESULTS The considerable amount of Cy A as a secondary metabolite, was produced under above described conditions, proved A. terreus as a potential source. The considerable amount of Cy A as a secondary metabolite, was produced under above described conditions, proved A. terreus as a potential source. Still no data on its use for Cy A production. Still no data on its use for Cy A production. It does used to extract lovastatin an agent used to lower cholesterol. It does used to extract lovastatin an agent used to lower cholesterol.


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