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Method for production of lipstatin through fermentation

A technology of lipox and statin, applied in the field of biopharmaceuticals, to achieve the effects of simple production and operation, low price and wide sources

Inactive Publication Date: 2013-01-16
PEKING UNIV FOUNDER GRP CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, the existing patent documents (such as EP0803576A2, US20050089978A1) on the fermentation process of liprestatin mainly focus on the optimization of the fermentation medium and the optimization of the extraction process, and there is no supplementary oxygen carrier in the fermentation process to improve the efficiency. Report on Dissolved Oxygen Concentration in Fermentation Broth of Prastatin

Method used

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  • Method for production of lipstatin through fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0049] 1. Preparation of methyl silicone oil feeding solution:

[0050] Accurately weigh 3 g (w / v) of methyl silicone oil, add 100 ml of purified water, stir, sterilize by high-pressure steam at 121°C for 15 minutes, and cool to obtain a sterile oxygen carrier feed solution. Prepare 5 parts of the same methyl silicone oil feeding solution.

[0051] 2. Ferment and cultivate Streptomyces toxin as follows:

[0052] The spores were inserted into 150ml seed culture medium, and the proportion of the culture medium was (w / v): 1.5% soybean powder, 2.5% glycerin, 1% soybean oil, and the balance was water; pH 7.0; sterilized at 121° C. for 30 minutes. The shaker rotates at 250 rpm, cultures at 28° C. for 20 hours, and transfers to 3 L of secondary seed medium, and the transfer amount is 5%.

[0053] The ratio of the secondary seed medium is (w / v): 2% soybean powder, 3% glycerin, 1.5% soybean oil, 0.1% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and the balance is water; pH...

Embodiment 2

[0058] 1. Preparation of hemoglobin feed solution:

[0059] Accurately weigh 0.3 g of hemoglobin, add 50 ml of sterile distilled water to dissolve, and filter to sterilize with a microporous membrane with a pore size of 0.25 μm.

[0060] Prepare 5 parts of the above hemoglobin solution for later use.

[0061] 2. Ferment and cultivate Streptomyces toxin as follows:

[0062] The spores were inserted into 150ml seed culture medium, and the proportion of the culture medium was (w / v): 1.5% soybean powder, 2.5% glycerin, 1% soybean oil, and the balance was water; pH 7.0; sterilized at 121° C. for 30 minutes. The shaker rotates at 250 rpm, cultures at 28° C. for 20 hours, and transfers to 3 L of secondary seed medium, and the transfer amount is 5%.

[0063] The ratio of the secondary seed medium is (w / v): 2% soybean powder, 3% glycerin, 1.5% soybean oil, 0.1% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and the balance is water; pH 7.0; 121 °C for 25 minutes. Stir at 3...

Embodiment 3

[0068] 1. Preparation of carbon tetrafluoride feeding solution:

[0069] Accurately weigh 5 g of carbon tetrafluoride, add 100 ml of purified water, stir, sterilize by high-pressure steam at 121° C. for 15 minutes, and cool to obtain a sterile oxygen carrier feeding solution. Prepare 5 parts of the same carbon tetrafluoride feeding solution.

[0070] 2. Ferment and cultivate Streptomyces toxin as follows:

[0071] The spores were inserted into 150ml seed culture medium, and the proportion of the culture medium was (w / v): 1.5% soybean powder, 2.5% glycerin, 1% soybean oil, and the balance was water; pH 7.0; sterilized at 121° C. for 30 minutes. The shaker rotates at 250 rpm, cultures at 28° C. for 20 hours, and transfers to 3 L of secondary seed medium, and the transfer amount is 5%.

[0072] The ratio of the secondary seed medium is (w / v): 2% soybean powder, 3% glycerin, 1.5% soybean oil, 0.1% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and the balance is water; ...

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Abstract

The invention provides a method for production of lipstatin through fermentation. The method includes: 1) conducting fermentation culture on the Streptomyces toxytricini used for lipstatin production in a fermentation medium; and 2) after 16-120 hours of the initiation of the fermentation cultivation in step 1), replenishing the obtained fermentation solution with aseptic oxygen-vectors, wherein the oxygen-vectors contain hemoglobin, methyl silicone oil or carbon tetrafluoride. Compared with processes without oxygen-vectors added, the invention makes the fermentation unit of lipstatin improved by more than 15%. At the same time, the method provided in the invention has the advantages of: simple production operation, low requirement for equipment, and low cost, thus being suitable for industrialized production.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals. Specifically, the present invention relates to a method for microbial fermentation to produce medicines, in particular to a method for fermentation to produce liprestatin. Background technique [0002] Lipstatin (lipstatin) is produced by fermentation of Streptomyces toxytricini, and its structure is shown in formula (I). It is hydrogenated to synthesize a new type of weight-loss drug orlistat, and its structure is shown in formula (II) ) shown. Orlistat is a long-acting and potent specific gastrointestinal lipase inhibitor that inactivates the enzymes by forming a covalent bond with the active serine sites of gastric and pancreatic lipases in the lumen of the stomach and small intestine. Fat in food cannot be decomposed into free fatty acids, so fat cannot be absorbed and utilized, thereby reducing calorie intake and controlling body weight. Orlistat is the first gastrointestinal lipase inhi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/02C12R1/465
Inventor 刘省伟
Owner PEKING UNIV FOUNDER GRP CO LTD
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