A kind of Lipistatin fermentation method and fermentation medium

A fermentation method and the technology of Lipus are applied in the field of Lipus statin fermentation method and fermentation medium, which 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 convenient and simple control. , to achieve large-scale production, the effect of improving the level of fermentation

Active Publication Date: 2022-07-22
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 in the following proportions: 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 min, and prepared into plate medium and slant medium for use. The spore glycerol tube of prestatin production strain stored in liquid nitrogen ultra-low temperature was diluted with sterile physiological saline and then coated on the above-mentioned plate medium, cultured for 12 days under the conditions of temperature of 28 ° C and humidity of 40-50%. Good, spore plump, uniform color single colonies were inoculated into the slant medium, and cultured for 8 days; from the above-mentioned mature test tube slants, the slant surfaces were all covered with uniform, white and dense bacterial layers and placed at 4°C Store in refrigerator.

[0067] b. Strain slanted shake flask fermentation screening: configure shake flask see...

Embodiment 2

[0075] The medium and culture conditions of plate separation and slant culture, strain slant shake flask fermentation screening, shake flask seed culture, primary seed tank culture and secondary seed tank culture are the same as in Example 1, and the fermenter culture medium formula and dissolved oxygen rebound The pre-cultivation conditions were the same as those in Example 1. The DO rebound time was 29h. After the rebound, the air volume and rotational speed were adjusted to control the dissolved oxygen. The dissolved oxygen began to drop to zero at 89h. The volume of each replenishment is 5% of the volume of the fermentation broth, the fermentation period is 161h, and the pot titer is 13570ug / ml.

Embodiment 3

[0077] The medium and culture conditions of plate separation and slant culture, strain slanted shake flask fermentation screening, shake flask seed culture, and first-level seed tank culture are the same as in Example 1. The sunflower oil was replaced with the same amount of soybean oil, the secondary seed tank culture conditions were the same as those in Example 1, the culture conditions before the fermentor dissolved oxygen rebound were the same as those in Example 1, and the dissolved oxygen DO rebound time was 32h, and after the rebound, according to the method in Example 1 Adjust the air volume and rotation speed to control the dissolved oxygen, and the dissolved oxygen starts to drop to zero at 116h. The water is replenished once at 116h, 140h and 164h. The volume of each replenishment is 5% of the volume of the fermentation broth. The fermentation period is 187h. The potency is 11890ug / ml.

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PUM

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Abstract

In order to overcome the problems of low fermentation unit, high cost of fermentation raw materials, complex and difficult process, and unsuitability for large-scale industrial production in the existing lipastatin fermentation methods, the present invention provides a simple and efficient fermentation method for lipastatin. and fermentation medium, including the following steps: plate separation and slant culture, bacterial strain slant shake flask fermentation screening, shake flask seed liquid preparation, primary seed tank culture, secondary seed tank culture and fermenter culture. In the present invention, the lipistatin fermentation method and fermentation medium obtained by combining optimization strategies such as screening of high-yielding strains by shaking flask fermentation, optimization of medium at all levels, and improvement of fermentation process technology have the advantages of high fermentation unit, low cost of fermentation raw materials, and control of fermentation process. The advantages of simplicity and convenience, no need to add exogenous toxic precursors, etc., are of great significance for improving lipstatin fermentation yield and product quality, and reducing production costs.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a lipistatin fermentation method and a fermentation medium. Background technique [0002] Lipistatin 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 lumen of the stomach and small intestine to inactivate 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 weight control, hypoglycemic, hypotensive, lipid-regulating and other purposes. Orlistat is the only over-the-counter drug approved by the US FDA, European EMA, and China's NMPA for the treatment of obesity. [0003] ...

Claims

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Application Information

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