Streptomyces toxytricini for high yield of lipstatin and fermentation medium thereof

A technology of Streptomyces toxin and fermentation medium, which is applied in the field of fermentation engineering, can solve the problems of low titer of fermentation products and restrictions on the industrial production of orlistat, and achieve the effect of improving high production capacity and promoting industrial production

Inactive Publication Date: 2015-07-22
BIOTOOL LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of fermentation to prepare liprestatin still faces the problem of low titer of fermentation products, which seriously restricts the industrial production of orlistat

Method used

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  • Streptomyces toxytricini for high yield of lipstatin and fermentation medium thereof
  • Streptomyces toxytricini for high yield of lipstatin and fermentation medium thereof
  • Streptomyces toxytricini for high yield of lipstatin and fermentation medium thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 starting strain mutagenesis

[0026] The starting strain was streptomyces toxytricini ATCC19813.

[0027] Preparation of spore suspension: In a sterile operating bench, insert streptomyces toxytricini ATCC19813 into solid plate medium (soluble starch 10g / L, KNO 3 2g / L, NaCl2g / L, K 2 HPO 4 2g / L, agar 2.2g / L, pH 7.2, sterilized at 0.1MPa at 121°C for 15min), after static culture at 28°C for 48h, use an inoculation loop to take the spores on the surface of the medium, transfer them into sterile water, and rotate and shake for 10min.

[0028] 1.1 Ultraviolet mutagenesis screening of soybean oil-tolerant strains

[0029] Put the spore suspension in an ultraviolet mutagenesis box (ultraviolet lamp power 30W, wavelength 253nm, direct vertical distance 30cm) for mutagenesis treatment for 40s, wrap it in black cloth, make gradient dilution with sterile water, spread it on a plate, and keep it away from the environment at 30°C. After 5-6 days of light culture, se...

Embodiment 2

[0036] The optimization of carbon source in the fermentation medium of embodiment 2

[0037] The carbon source is one of the main raw materials for the composition of the medium. Its main function is to provide the ability for the growth and reproduction of microorganisms, and to provide the required carbon components for the synthesis of bacteria and the synthesis of target products. Different kinds of carbon sources were added to the fermentation medium, and the significant differences of different carbon sources on the growth of microorganisms and the product synthetic oil were investigated.

[0038] Seed medium: soybean cake powder 20g / L, glycerin 20g / L, yeast extract 7.5g / L, soybean oil 5g / L, KNO 3 2g / L, MgSO 4 7H 2 O1g / L, pH7.0‐7.2, 0.1MPa sterilization at 121℃ for 15min.

[0039] Fermentation medium: soybean cake powder 40g / L, carbon source (for specific types of carbon source, see figure 1 ) 30g / L, soybean oil 50g / L, lecithin 10g / L, yeast powder 40g / L, (NH 4 ) 2 ...

Embodiment 3

[0042] The optimization of nitrogen source in the fermentation medium of embodiment 3

[0043] As one of the main components of the culture medium, the nitrogen source is mainly used for the cellular substances that constitute the bacteria, such as amino acids, proteins, nucleic acids, etc., and also provides nitrogen components for the synthesis of target products. The compound nitrogen source composed of yeast nitrogen source and non-yeast nitrogen source in the medium is beneficial to the synthesis of fermentation products. Different kinds of yeast nitrogen sources and non-yeast nitrogen sources were added to the fermentation medium, and the significant differences of different nitrogen sources on microbial growth and product synthetic oil were investigated.

[0044] Seed medium and fermentation conditions are the same as in Example 2.

[0045] In the fermentation medium (non-yeast nitrogen source 40g / L, lecithin 10g / L, glycerol 30g / L, soybean oil 50g / L, (NH 4 ) 2 SO 4 ...

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Abstract

The invention provides streptomyces toxytricini for high yield of lipstatin, wherein the streptomyces toxytricini can high-efficiency utilize grease, and has a performance of high yield of lipstatin. On the other hand, the invention further provides an optimized streptomyces toxytricini fermentation medium.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, and in particular relates to a strain of Streptomyces toxin, which has the performance of high-yielding riprestatin. The invention also relates to a fermentation medium for the strain. Background technique [0002] Liprestatin is an endogenous lipase inhibitor produced by streptomyces toxytricini, and its tetrahydro derivative orlistat is currently the only weight-loss drug marketed as a non-central nervous system effect [1] . Orlistat has been certified by the FDA and currently occupies 2 / 3 of the weight loss drug market. Orlistat can selectively inhibit the activity of pancreatic lipase in the intestinal tract. As a "fat blocking agent", it only exerts pharmacological effects in the gastrointestinal tract and can prevent about 30% of dietary triglycerides from being digested and absorbed , On the basis of food therapy, calorie intake can be further reduced. Orlistat's high selectivit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/02C12R1/465
Inventor 罗煜丁时诚瞿旭东
Owner BIOTOOL LLC
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