Method for fermentation production of lipstatin

A technology of liprex and statin, which is applied in the field of fermentative production of liprestatin, can solve the problems of reduced output of metabolites, shortened product synthesis stages, and consumption, and achieves the effects of reducing fermentation costs and increasing fermentation titer

Active Publication Date: 2014-07-23
鲁南新时代生物技术有限公司
View PDF10 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the commonly used feeding methods are periodic feeding or continuous constant-speed feeding. Since periodic feeding is a one-time feeding of carbon sources, nitrogen sources and other necessary substrates, the concentration of nutrients will be too high in a short period of time. This will cause the bacteria to develop in the direction of a large number of mycelia, so that the nutrients are mainly consumed on the growth of the mycelium, the synthesis stage of the product is shortened, and the output of the final metabolite is reduced.
Although the continuous constant-rate feeding method can eliminate the phenomenon of suppressing mycelia growth due to the high concentration of nutrients caused by the one-time feeding of a large amount of nutrients in periodic feeding, there are still certain defects in this feeding method, that is, in the whole feeding The same amount of substrate is added at a constant rate during the feed cycle, while ignoring the actual consumption of the bacteria in each stage of the metabolic process

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for fermentation production of lipstatin
  • Method for fermentation production of lipstatin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024]Slant culture: Prepare medium according to the following ratio: glucose 2g / L, malt extract 8g / L, yeast extract 10g / L, agar 15g / L, pH 7.1. Prepare 300 mL of culture medium, heat the agar to melt, stir evenly, put it into 10 large test tubes, sterilize, and make it into a slope for later use. Dissolve the freeze-dried Streptomyces toxin strains in 5 mL of sterile water, take 4 mL and evenly spread them on 10 inclined surfaces at a temperature of 28°C and a humidity of 40%, and cultivate them for 10 days to produce a large number of spores. spore suspension.

[0025] Shake flask seed solution preparation: Prepare medium according to the following ratio: glycerol 10g / L, malt extract 10g / L, yeast extract 5g / L, defatted soybean powder 20g / L, pH 6.0. Prepare 200mL of culture medium, put it into 4 bottles of 500mL Erlenmeyer flasks (each bottle contains 50mL), sterilize and set aside. Inoculate 2 mL of the spore suspension into the shake flask. The temperature was 28° C. and ...

Embodiment 2

[0029] With reference to Example 1, the slant culture was carried out, the shake flask seed solution was prepared, and the seed tank was cultivated. Fermentation tank culture: a 50L fermenter was used for cultivation. Among them, the liquid medium was prepared according to the following proportions: glycerin 10g / L, defatted soybean powder 20g / L, soybean oil 10g / L, antifoaming agent 2g / L, pH 7.0. The volume after elimination is 33.25L. When the temperature drops to 28°C, the cultured seed tank bacteria are inoculated into the fermenter with an inoculum size of 5% (1.75L). Under the condition of 28°C, the whole process Control the dissolved oxygen at 40% by adjusting tank pressure, rotation speed and ventilation. The stirring speed is 300-540 rpm, the ventilation rate is 0.6VVM-1.1VVM (volume / volume / minute), and the tank pressure is 0.06-0.08MP. From 29 hours after fermentation and inoculation, the dissolved oxygen rose sharply to 70%, and immediately began to feed the mixed f...

Embodiment 3

[0031] With reference to Example 1, the slant culture was carried out, the shake flask seed solution was prepared, and the seed tank was cultivated. Fermentation tank culture: 500L fermentation tank was used for cultivation. Among them, the liquid medium was prepared according to the following proportions: glycerol 10g / L, defatted soybean powder 20g / L, soybean oil 10g / L, defoamer 2g / L, pH 7.0. The volume after elimination is 335L. When the temperature drops to 28°C, the cultured seed tank bacteria are inoculated into the fermenter with an inoculum size of 5% (16.7L). Under the condition of 28°C, the whole process Control the dissolved oxygen at 40% by adjusting the tank pressure, rotation speed and ventilation. The stirring speed is 300-540 rpm, the ventilation rate is 0.6VVM-1.1VVM (volume / volume / minute), and the tank pressure is 0.06-0.08MP. From 30 hours after fermentation and inoculation, the dissolved oxygen increased sharply to 70%, and immediately began to feed the mi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention relates to a method for fermentation production of lipstatin, particularly to application of a mixed fatty acid to replace linoleic acid supplemented in a fed-batch manner during a fermentation culture process, wherein the mixed fatty acid comprises, by weight, 2.0-7.0% of stearic acid, 14.0-30% of oleic acid, 61-78% of linoleic acid, and 5.0-7.0% of palmitic acid. According to the present invention, on the basis of the existing invention CN102268466, the mixed fatty acid is adopted to replace the linoleic acid supplemented in the fed-batch manner during the fermentation culture process, such that the fermentation cost is reduced, and the HPLC detection results show that the fermentation broth does not contain the methionine analog lipstatin; and with the variable speed material supplementing method, nutrient balance is always maintained in the culture medium, the sufficient and non-excess nutrients are provided at the lipstatin synthesis stage, and the fermentation titer is improved by 6-16%.

Description

technical field [0001] The invention relates to a method for fermenting and producing riprestatin, belonging to the field of microbial fermentation. Background technique [0002] Liprestatin (Formula 1) is an intermediate of the new weight loss drug orlistat. 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 To play a therapeutic role, inactivated enzymes cannot hydrolyze the fat in food, mainly triglycerides, into absorbable free fatty acids and monoacylglycerols. Undigested triglycerides cannot be absorbed by the body, thereby reducing caloric intake and controlling weight. [0003] [0004] There are two methods for the production of orlistat, fermentation synthesis and total synthesis. The production cost of the total synthesis method is relatively high. The production c...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12P17/02C12R1/465
Inventor 张太凤任显江倪伟东
Owner 鲁南新时代生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap