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Streptomyces toxytricini and applications of streptomyces toxytricini in preparation of lipstatin

A technology of Streptomyces toxin and statin, applied in the direction of bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of unstable production capacity, inability to industrialize production, low yield, etc., 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: 2015-02-11
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

Method used

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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
Comparison scheme
Effect test

Embodiment １

[0027] Example 1: Culture characteristics of Streptomyces toxins ST-5423

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

[0029]Table 1, the bacterial colony characteristics of the strain of Streptomyces toxin Streptomyces ST-5423 on various media

[0030] culture medium

Embodiment 2

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

[0032] Physiological and biochemical properties were performed using standard methods in the classification of Streptomyces toxins.

[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 strains were 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 inoculation solution (IF-C) to make a bacterial suspension, and adjust to 98% T / FF with a turbidimeter. Use an 8-hole electric pipette to add the bacterial suspension to each well of the Biolog (GENⅢ) microwell, 100 μl per well. Place the microwell identification plate in a 28°C incubator, and place it on a Biolog reader to read the results...

Embodiment 3

[0041] Embodiment 3: the production fermentation culture of liprestatin

[0042] 1. Seed culture and storage

[0043] Solid medium: glucose 5g, yeast extract 5g, malt extract powder 5g, magnesium sulfate 0.2g, potassium dihydrogen phosphate 0.5, agar 18g, add water 1000ml, pH7.0

[0044] Cultivation: the strain is inoculated on a solid medium slant, and cultured at 28°C for 7-10 days.

[0045] After the solid culture is over, place the slant at 4°C and refrigerate for later use.

[0046] 2. shake flask seed culture

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

[0048] Culture: 26°C, 28h, 200rpm

[0049] When the hyphae grow up, there is obvious wall-hanging phenomenon and the bacterial concentration is about 9%. Put the 2L shake flask seed culture solution into the following seed tanks.

[0050] 3. seed pot seed culture

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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 present invention relates to a novel microorganism and its use and application, in particular to a streptomyces toxin and its application in the preparation of riprestatin. Background technique [0002] Liprestatin is a fermentation product. It is cultured in the fermentation broth of Streptomyces toxytricini to obtain a fermentation filtrate, which is then extracted and concentrated to obtain a concentrated solution of liprestatin. After a series of purification processes such as hydrogenation, crystallization, and centrifugation And so can finally obtain the finished product of orlistat. And orlistat is a long-acting and potent specific gastrointestinal lipase inhibitor, it forms a covalent bond with the active serine site of gastric lipase and pancreatic lipase in the lumen of stomach and small intestine to deactivate the enzyme Live to play a therapeutic role, the inactivated enzymes cannot hydrolyze the fat in food, mainly triglycerides, into...

Claims

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Application Information

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Patent Type & Authority Patents(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