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Cyclosporins A fermentation production method

A cyclosporin and production method technology, applied in the field of cyclosporine A fermentation technology, can solve the problems of high medium requirement and difficulty in producing cyclosporin A, prolong the fermentation cycle and overcome the synthesis rate of bacteria Too fast, the effect of increasing the content

Active Publication Date: 2005-09-21
SHANGHAI INST OF PHARMA IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This not only provides the possibility to search for new cyclosporine compounds, but at the same time causes certain difficulties to produce high-component cyclosporine A, because it has relatively high requirements on the medium

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Fusarium solan ATCC 46829. Slant culture is carried out, and the weight composition of the medium is: corn flour 3.0%; glucose 2.0%; agar 2.0%, the balance is water; pH 6.5; temperature 25°C, culture time 7 days;

[0038] (2) Seed cultivation:

[0039] The bacterial classification is then cultivated under the following medium and culture conditions:

[0040] Medium weight composition: corn flour 3.0%; glucose 2.0%; KH 2 PO 4 0.2%; KCl0.05%; MgSO 4 .7H 2 O 0.005%; NaNO 3 0.05%, the balance is water;

[0041] pH 6.5; temperature 25°C; filling volume 20ml / 250ml; culture time 2 days;

[0042] (3) Inoculate the strains for fermenting and producing cyclosporin A in the culture medium, the inoculum size is 7%, and then ferment and cultivate;

[0043] The technological conditions of fermentation culture are:

[0044] Medium weight composition: corn flour 3.0%; glucose 3.0%; flour 4.0%; casein 1.2%; MgSO 4 .7H 2 O 0.005%; KH2 PO 4 0.2%;NaNO 3 0.15%; KCl0.05%...

Embodiment 2

[0050] Adopt the same method of embodiment 1, the filling capacity of seed culture and fermentation culture is 50ml / 750ml;

[0051] Fermentation culture medium weight composition: corn flour 3.0%; glucose 3.0%; flour 4.0%; casein 0.6%; MgSO 4 .7H 2 O 0.005%; KH 2 PO 4 0.2%;NaNO 3 0.15%; KCl0.05%; CaCO 3 0.2%, the balance is water;

[0052] Adopt aforementioned analytical test method to detect, the result is as follows:

[0053] The fermentation unit of cyclosporin A is 850 μg / ml; the content of active components of cyclosporin A is 69%.

Embodiment 3

[0055] Cyclosporin A aerated culture studies:

[0056] In the optimization experiment of laboratory fermentation conditions, the effect of aeration on CyA biosynthesis was firstly studied. It was found that aeration had a great influence on the biosynthesis of CyA. Table 1 shows the biosynthesis of CyA and other relevant parameters of cyclosporin A producing bacteria under the conditions of 25°C, culture time of 4 days, and medium load of 50ml / 750ml, 100ml / 750ml, 150ml / 750ml and 200ml / 750ml respectively metabolism.

[0057] Table 1 Metabolism of CyA-producing bacteria biosynthesis and other related parameters under different aeration conditions

[0058] Loading capacity CyA titer CyA content Mycelium pH Total sugar Reducing sugar Amino acid

[0059] (ml) (μg / ml) (area%) amount (mg / ml) (mg / ml) (mg / ml)

[0060] 50 892 86.1 35 5.63 1.31 0.10 31.1

[0061] 100 963 87.7 39 5.47 2.32 0.29 35.1

[0062] 150 654 83.3 45 5.49 2.55 0.42 35.3

[0063] 200 321 84.7 42 5.87 3.74 0.4...

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PUM

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Abstract

The invention belongs to the microbial drug technology in the field of biomedicine. The invention provides a novel production process of cyclosporine A raw material fermentation. The process of the present invention adopts new medium components and concentrations, and new culture conditions, so that the biosynthesis period of cyclosporine A producing bacteria is extended, and the foam problem that occurs in the large-scale production process is overcome, so that feeding can be used process, finally at a production scale of 20 tons, the fermentation unit of cyclosporin A in the fermentation broth increased by 100%, the content of effective components increased by about 7%, the volume of the tank increased by more than 27%, and the total tank volume increased by more than 159%. And the fermentation index is increased by more than 82%. There is no significant difference in the quality of the finished product, and the specifications meet the standards.

Description

technical field [0001] The invention relates to a cyclosporine A fermentation process. Background technique [0002] Cyclosporin A (cyclosporin A, CyA) was first isolated from the soil of the United States and Norway by the Swiss Sandoz Company in 1970 when screening antifungal drugs at the Basel Institute of Microbiology. Cylindrocarpon lucidum and Beauveria bassiana produce cyclosporine A with antifungal activity. In 1974, the immune activity research in animals was carried out, and in 1978, it was first used in clinical kidney transplantation trials. In 1980, the total synthesis of cyclosporine A was realized. In 1983, the US FDA approved it for clinical organ transplantation. Its detailed information has been publicly reported in Process Biochemistry 1991, 26: 157-166. [0003] As an immunosuppressant, the main feature of cyclosporin A is the selective inhibitory effect, that is, it mainly inhibits the proliferation and differentiation of T cells. Cyclosporin A inhi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12PC12P21/02C12P21/04
Inventor 陈代杰吴晖李继安戈梅吴萍吴飞夏云飞
Owner SHANGHAI INST OF PHARMA IND CO LTD
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