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Method for extracting lipstatin from fermentation broth with one-pot process

A fermented liquid and statin technology, applied in the field of medicine, can solve the problems of lengthy steps, unspecified content and yield, etc., and achieve the effects of improving extraction yield, reducing extraction and purification steps, and reducing production costs

Inactive Publication Date: 2015-11-18
LIAOCHENG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method steps are tedious, and the content and yield are not stated

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] (1) After the fermentation of riprestatin by Streptomyces toxins, transfer 15L of fermentation broth to a 50L multifunctional glass tank, add dilute hydrochloric acid to adjust the pH to 1.5, heat to 45°C, keep for 30min, and cool to room temperature.

[0010] (2) Add 1 / 2 volume of heptane into the glass jar, stir at 150 rpm, extract for 30 minutes, stand still for phase separation, and remove the raffinate phase from the bottom valve to obtain the heptane phase.

[0011] (3) Add 1 times the volume of 2.5% sodium chloride solution, stir and wash for 5 minutes, let stand for stratification, remove the lower aqueous phase, repeat several times until the lower layer is clear and translucent, and obtain the upper heptane phase.

[0012] (4) Concentrate the heptane phase under reduced pressure to 1 / 3 volume, and concentrate under reduced pressure at a temperature of 50°C. After the concentration, add 1 times the volume of pure water, and repeat washing until the lower layer i...

Embodiment 2

[0015] (1) After the fermentation of liprestatin by Streptomyces toxins, transfer 15L of fermentation broth to a 50L multifunctional glass tank, add dilute hydrochloric acid to adjust the pH to 2.0, heat to 65°C, keep for 30min, and cool to room temperature.

[0016] (2) Add 1 / 2 volume of heptane into the glass jar, stir at 200 rpm, extract for 30 minutes, stand still for phase separation, and remove the raffinate phase from the bottom valve to obtain the heptane phase.

[0017] (3) Add 1 times the volume of 5% sodium chloride solution, stir and wash for 10 minutes, let stand to separate layers, remove the lower aqueous phase, repeat several times until the lower layer is clear and translucent, and obtain the upper heptane phase.

[0018] (4) Concentrate the heptane phase under reduced pressure to 1 / 3 volume, and concentrate under reduced pressure at a temperature of 50°C. After the concentration, add 1 times the volume of pure water, and repeat washing until the lower layer is...

Embodiment 3

[0021] (1) After the fermentation of riprestatin by Streptomyces toxins, transfer 15L of fermentation broth to a 50L multifunctional glass tank, add dilute hydrochloric acid to adjust the pH to 3.0, heat to 55°C, keep for 30min, and cool to room temperature.

[0022] (2) Add 1 / 2 volume of heptane into the glass jar, stir at 200 rpm, extract for 30 minutes, stand still for phase separation, and remove the raffinate phase from the bottom valve to obtain the heptane phase.

[0023] (3) Add 1 times the volume of 3% sodium chloride solution, stir and wash for 10 minutes, let stand to separate layers, remove the lower aqueous phase, repeat several times until the lower layer is clear and translucent, and obtain the upper heptane phase.

[0024] (4) Concentrate the heptane phase under reduced pressure to 1 / 2 volume, and concentrate under reduced pressure at a temperature of 55°C. After the concentration, add 1 times the volume of pure water, and repeat washing until the lower layer is...

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Abstract

The invention discloses a method for extracting lipstatin from a fermentation broth with a one-pot process. After fermentation of lipstatin by using streptomyces toxytricini ends, the fermentation broth is transferred to a multifunctional extraction tank, inorganic acid is added, the pH (potential of hydrogen) is adjusted to be acidic, and the mixture is heated and then cooled to the normal temperature; heptane is added for extraction, the mixture is left to stand for phase splitting, and a heptane phase is obtained; a sodium chloride solution with the volume equal to that of the mixture is added to wash the mixture and left to stand for layering, and the heptane phase is obtained; concentration under the reduced pressure is performed until the volume is reduced to 1 / 3-1 / 2 of the original volume, purified water is added, the product is washed for multiple times, concentration under the reduced pressure is performed continuously until the volume is reduced to be 1 / 6-1 / 4 of the original volume, cooling crystallization and filter pressing at the low temperature are performed, and lipstatin with the content not lower than 80% is obtained.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for purifying lipstatin. Background technique [0002] Lipstatin is a metabolite of Streptomycestoxytricini. It is an inhibitor of pancreatic lipase in the gastrointestinal tract of the human body. It reduces the decomposition and absorption of high-fat food by the human body. It can be used as a weight loss drug itself, but Because lipstatin is an unsaturated fatty acid, it is easily oxidized and unstable in the air. Roche synthesized lipstatin into tetrahydro derivatives through hydrogenation reaction, which is the famous weight-loss drug orlistat. [0003] At present, there are two production processes for orlistat: one is the total chemical synthesis method, which has a long process route, low yield and high cost; Fermentation produces lipstatin, which is then chemically hydrogenated to produce orlistat. In the microbial semi-synthesis method, the cost...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D305/12
CPCC07D305/12
Inventor 董惠钧李荣荣王素珍
Owner LIAOCHENG UNIV
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