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Lipstatin fermentation method and fermentation medium

A fermentation method and culture medium technology, applied in the directions of fermentation, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of unsuitability for large-scale industrial production, complex and difficult fermentation process, and high cost of fermentation raw materials, and achieve control It is convenient and simple, realizes the effect of large-scale production and improved fermentation level

Active Publication Date: 2021-05-28
ARGUS PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem solved by the present invention is to provide a fermentation unit with high fermentation unit, high fermentation raw material cost, complex and difficult to control fermentation process, and unsuitable for large-scale industrial production in the existing lipstatin fermentation method. A lipstatin fermentation method and fermentation medium suitable for industrial production with low cost of raw materials, simple and convenient fermentation process control, and no need to add exogenous toxic precursors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] a. Plate separation and slant culture: configure the spore medium according to the following ratio: glucose 4g / L, yeast extract 4g / L, malt extract 10g / L, calcium carbonate 2g / L, agar powder 18g / L, add alkali to adjust pH to 6.75, sterilized at 115°C for 30 minutes, and made into plate medium and slant medium for later use. Dilute the spore glycerol tubes of prastastatin-producing strains stored in liquid nitrogen cryogenically with sterile saline, spread them on the above-mentioned plate culture medium, and cultivate them for 12 days at a temperature of 28°C and a humidity of 40-50%. Inoculate the slant culture medium with a single colony that is good, full of spores, and uniform in color, and continue to cultivate for 8 days; select the slant surface that is completely covered with a uniform, white and dense bacterial layer from the above mature test tube slant and place it at 4 °C. Store in the refrigerator.

[0067] b. Slant shake flask fermentation screening of str...

Embodiment 2

[0075] Plate separation and slant culture, bacterial strain slant shake flask fermentation screening, shake flask seed culture, primary seed tank culture and secondary seed tank culture medium and culture conditions are the same as in Example 1, fermenter medium formula and dissolved oxygen rebound Pre-cultivation conditions are the same as in Example 1, the dissolved oxygen DO rebound time is 29h, after the rebound, adjust the air volume and rotating speed to control the dissolved oxygen according to the method in Example 1, and the dissolved oxygen starts to drop to zero at 89h after fermentation, and water is replenished once at 89h, 113h and 137h. The volume of water added each time is 5% of the volume of the fermentation broth, the fermentation period is 161 hours, and the tank titer is 13570ug / ml.

Embodiment 3

[0077] Plate separation and slant culture, bacterial strain slant shake flask fermentation screening, shake flask seed culture, medium and culture conditions of one-level seed tank culture are the same as embodiment 1, and the culture medium in the second-level seed tank and fermenter culture process will be Sunflower oil is replaced by an equivalent amount of soybean oil, the secondary seed tank culture conditions are the same as in Example 1, the culture conditions in the fermenter before the dissolved oxygen rebound are the same as in Example 1, the dissolved oxygen DO rebound time is 32h, and after the rebound, follow the method in Example 1 Adjust the air volume and speed to control the dissolved oxygen. Ferment until 116h and the dissolved oxygen starts to drop to zero. Water is replenished once at 116h, 140h and 164h. The volume of each replenishment is 5% of the volume of the fermentation broth. The potency of 187h in the fermentation cycle is 11890ug / ml.

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Abstract

In order to overcome the problems of low fermentation unit, high cost of fermentation raw materials, complex and difficult control of process, unsuitability for large-scale industrial production and the like in the existing fermentation method of the lipstatin, the invention provides a simple, convenient and efficient fermentation method of the lipstatin and a fermentation culture medium. The method comprises the following steps: plate separation and slant culture, strain slant shake flask fermentation screening, shake flask seed liquid preparation, primary seed tank culture, secondary seed tank culture and fermentation tank culture. The fermentation method and the fermentation culture medium of the lipstatin, which are obtained through combined optimization strategies such as shake flask fermentation screening of high-yield strains, optimization of culture media at all levels and improvement of a fermentation process technology, have the advantages of high fermentation unit, low fermentation raw material cost, simplicity and convenience in fermentation process control, no need of adding exogenous toxic precursors and the like; and the method has very important significance for improving the fermentation yield and the product quality of the lipstatin and reducing the production cost.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a lipstatin fermentation method and a fermentation medium. Background technique [0002] Lipstatin is a natural product produced by Streptomyces toxytricini with good lipase inhibitory activity, and its hydrogenated derivative orlistat is currently the best-selling weight-loss drug in the world. Orlistat, as a gastrointestinal lipase inhibitor, forms a covalent bond with the active serine site of lipase in the stomach and small intestine, inactivating lipase so that it cannot decompose fat in food into absorbable free fatty acids and monoacylglycerols, thereby reducing calorie intake, controlling weight gain, and achieving the purpose of weight control, lowering blood sugar, lowering blood pressure, and regulating fat. Orlistat is the only over-the-counter drug approved by the US FDA, the European EMA, and the Chinese NMPA for the treatment of obesity....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/02C12N1/20C12R1/465
CPCC12P17/02C12N1/20
Inventor 郭霞凌李辉
Owner ARGUS PHARMA
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