Streptomyces toxytricini and applications of streptomyces toxytricini in preparation of lipstatin

A technology of streptomyces and statins, which is applied in bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of unstable production capacity, inability to industrialize production, and low yield, and achieve extraction and purification. The process is simple, the yield and purity are improved, and the fermentation by-products are less.

Active Publication Date: 2013-12-25
HANGZHOU HUADONG MEDICINE GRP PHARMA RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] General liprestatin-producing bacteria have unstable production capacity, low yield, more fermentation by-products, and more impurities, which lead to more complicated extraction, hydrogenation, and purification processes into orlistat in the later stage, which greatly increases the production cost of orlistat. It is difficult to purify the sequence, and it is difficult to obtain a high-purity final product, so it cannot be industrialized

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  • Streptomyces toxytricini and applications of streptomyces toxytricini in preparation of lipstatin
  • Streptomyces toxytricini and applications of streptomyces toxytricini in preparation of lipstatin

Examples

Experimental program
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Effect test

Embodiment １

[0027] Example 1: The culture characteristics of Streptomyces toxotris ST-5423

[0028] The strain of Streptomyces toxotris ST-5423 was inoculated on glycerol asparagine agar, yeast extract malt extract agar and other media for observing morphological characteristics. Incubate at 28°C for 5, 8, 10 days, observe and describe. The results are shown in Table 1.

[0029] Table 1. Colony characteristics of strains of Streptomyces toxotris ST-5423 on various media

[0030] Medium

Embodiment 2

[0031] Example 2: Carbon source utilization and physiological and biochemical characteristics

[0032] Physiological and biochemical properties adopt the standard method in the classification of Streptomyces toxin.

[0033] Using the Biolog (GENⅢ) automatic microbial identification system, 94 phenotypic tests were performed on the strain Streptomyces toxytricini ST-5423, including 71 carbon source utilization tests and 23 chemical sensitivity tests: the strain was inoculated on BUG+B plate medium Incubate at a constant temperature of 28°C for 16 hours, wash the bacteria on the plate with a sterile cotton swab, mix with the inoculum (IF-C) to make a bacterial suspension, and adjust to 98% T / FF with a turbidimeter. Use an 8-hole electric liquid feeder to add the bacterial suspension to each well of the Biolog (GENⅢ) micropores, 100μl per well. Place the microwell identification plate in a 28°C incubator, and place it on a Biolog reader to read the results after culturing for 12h, 24...

Embodiment 3

[0041] Example 3: Production, fermentation and cultivation of lipristatin

[0042] 1. Seed strain cultivation and preservation

[0043] Solid medium: glucose 5g, yeast extract 5g, malt extract powder 5g, magnesium sulfate 0.2g, potassium dihydrogen phosphate 0.5, agar 18g, 1000ml of water, pH 7.0

[0044] Cultivation: The strains are inoculated on a solid medium slant and cultivated at 28°C for 7-10 days.

[0045] After the solid culture is completed, place it on a slope at 4°C and refrigerate for later use.

[0046] 2. Shake bottle seed culture

[0047] Medium: starch hydrolyzed sugar 20g, glycerin 60g, potato extract powder 10g, soybean meal powder 6g, vegetable peptone 4g, yeast powder 20g, MgSO4 2g, ZnSO4 6g, add 2L water, pH7.0.

[0048] Cultivation: 26°C, 28h, 200rpm

[0049] When the mycelium grows, there is obvious wall hanging phenomenon and the bacteria concentration is about 9%. Put 2L shake flask seed culture solution into the following seed tank.

[0050] 3. Seed pot seed ...

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Abstract

The invention discloses Streptomyces toxytricini ST-5423 and applications of Streptomyces toxytricini ST-5423. The bacterial strain is preserved in China Center for Type Culture Collection, the preservation number is CCTCC M2013312, and the preservation date is July, 4th, 2013. The invention also discloses a preparation method of an orlistat intermediate by fermentation using the bacterial strain. It is confirmed by production practices, in which 10t and 30t fermentation tanks are used, that production capacity of fermentation technology disclosed in the invention is stable, fermentation titer unit is high, fermentation by-products are relatively less, the difficulty of post-extraction is reduced greatly, the fermentation technology is suitable for industrialized large-scaled production, and quality of the obtained orlistat products is high.

Description

Technical field [0001] The invention relates to a new type of microorganism and its use and application, in particular to a toxotridin and its application in the preparation of lipstatin. Background technique [0002] Lipstatin is a fermentation product. It is cultured by Streptomyces toxytricini fermentation broth to obtain a fermentation filtrate, and then extracted and concentrated to obtain a lipstatin concentrate. After a series of purification processes such as hydrogenation, crystallization, and centrifugation The finished product of Orlistat can be finally obtained. Orlistat is a long-acting and potent specific gastrointestinal lipase inhibitor. It forms a covalent bond with the active serine sites of gastric lipase and pancreatic lipase in the stomach and small intestine to deactivate the enzyme. Live and play a therapeutic role, inactivated enzymes cannot hydrolyze fats in food, mainly triglycerides, into absorbable free fatty acids and monoacylglycerols. Undigested t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/02C12R1/465
Inventor 李立似方军蒋彩霞金利琴何金芳刘金国
Owner HANGZHOU HUADONG MEDICINE GRP PHARMA RES INST
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