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Method for producing lipstatin by virtue of fermentation

A technology of lipox and statin, which is applied in the field of fermentation biosynthesis of lipstatin, can solve the problems of complex operation and difficult control of the production method, difficult to remove impurities, low purity, low fermentation level, etc. Fully and thoroughly, the effect of increasing fermentation yield

Inactive Publication Date: 2015-05-20
HEBEI SHENGXUE DACHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention provides a method for producing lipstatin by fermentation in order to solve the technical problems of low fermentation level, difficult-to-remove impurities, low purity, complex operation and uncontrollable production method in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A. Preparation of slant strains: configure slant medium according to the following proportions: 0.8% soluble starch, 0.05% casein, 0.2% sodium chloride, 0.2% potassium dihydrogen phosphate, pH 6.0, and set aside. Inoculate the sand spores on the slant culture medium, cultivate them for 5 days at a temperature of 26°C and a humidity of 40%, to obtain slant spores;

[0026] B. Preparation of shake flask seeds: prepare shake flask seed culture medium according to the following proportions: 16% soybean cake powder, 15% glycerin, 0.3% linseed oil, 1.0% magnesium sulfate, pH 7.0, and set aside. The inclined spores cultivated in step A were washed with 10 mL of sterile physiological saline and inoculated into the shake flask seed medium, and cultivated for 20 hours at a rotation speed of 200 rpm and a temperature of 26° C. to obtain shake flask seeds;

[0027] C. Preparation of seed solution: Prepare seed medium according to the following ratio: bean cake powder 18%, glycerin ...

Embodiment 2

[0030] A. Preparation of slant strains: prepare slant culture medium according to the following proportions: 0.8% soluble starch, 0.05% casein, 0.2% sodium chloride, 0.2% potassium dihydrogen phosphate, pH 6.8, and set aside. Inoculate the sand spores on the slant culture medium, cultivate them for 7 days at a temperature of 28°C and a humidity of 60%, to obtain slant spores;

[0031] B. Preparation of shake flask seeds: prepare shake flask seed culture medium according to the following proportions: 16% of soybean cake powder, 15% of glycerol, 0.3% of linseed oil, 1.0% of magnesium sulfate, pH 7.2, and set aside. The slant spores cultivated in step A were washed with 10 mL of sterile physiological saline and inoculated into the shake flask seed medium, and cultivated for 31 hours at a rotation speed of 220 rpm and a temperature of 28° C. to obtain shake flask seeds;

[0032]C. Preparation of seed solution: Prepare seed medium according to the following ratio: 18% bean cake pow...

Embodiment 3

[0035] A. Preparation of slant strains: prepare slant medium according to the following proportions: 0.8% soluble starch, 0.05% casein, 0.2% sodium chloride, 0.2% potassium dihydrogen phosphate, pH 6.1, and set aside. Inoculate the sand spores on the slant medium, and cultivate them for 6 days at a temperature of 26.5°C and a humidity of 45 to obtain slant spores;

[0036] B. Preparation of shake flask seeds: prepare shake flask seed culture medium according to the following proportions: 16% soybean cake powder, 15% glycerin, 0.3% linseed oil, 1.0% magnesium sulfate, pH 7.0, and set aside. The slant spores cultivated in step A were washed with 10 mL of sterile physiological saline and inoculated into the shake flask seed medium, and cultivated for 23 hours at a rotation speed of 205 rpm and a temperature of 26.5° C. to obtain shake flask seeds;

[0037] C. Preparation of seed solution: Prepare seed medium according to the following ratio: bean cake powder 18%, glycerin 14%, li...

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Abstract

The invention discloses a method for producing lipstatin by virtue of fermentation. The method comprises the following steps: preparing an inclined plane strain of streptomyces toxytricini, preparing shake flask seeds, preparing a seed solution, performing fermentation and cultivation, and adding linseed oil in a fed-batch mode after performing fermentation and cultivation for 50 hours, wherein calculated based on the volume of a fermentation canned material, the fed-batch rate of the linseed oil is 0.03-0.2L / h / t. The method disclosed by the invention can be controlled and operated more easily by only adding one substance namely the linseed oil in a fed-batch mode after performing fermentation and cultivation for 50 hours than by replenishing a mixture of linoleic acid and leucine as well as a mixture of mixed fatty acid and a leucine solution; on the premise of early fed-batch fermentation, the measures of further improving the fermentation yield, more completely utilizing a culture medium and cooling after fermenting for 130 hours can be controlled, bacterium decay can be delayed by virtue of cooling, the fermentation cycle can be prolonged by 20-30 hours, and the production titer time can be increased; and moreover, the culture medium can be used more fully and completely, the pressure of three-waste treatment can be reduced, and the method can be more environment-friendly.

Description

technical field [0001] The invention relates to the technical field of lipstatin production, in particular to a fermentation biosynthesis of lipstatin. Background technique [0002] Lipstatin is a metabolite produced by the fermentation of Streptreptomyces toxytricini, and its stable hydrogenated product, orlistat, can be used as a lipase inhibitor to inhibit the activity of lipase in the intestine. Reduce the decomposition and absorption of fat in the intestinal tract, and play a role in weight loss. [0003] Due to the high cost of the chemical total synthesis method, it is mainly produced by the biological fermentation method at present, which is relatively low in cost and easy to produce. At present, it has been reported that the process of supplementing precursors includes supplementing linoleic acid, leucine, octanoic acid and other precursor substances, which can increase the potency, but in subsequent studies, it was found that by supplementing precursors such as li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/02C12R1/465
Inventor 郭耀光刘丽芳
Owner HEBEI SHENGXUE DACHENG PHARMA
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