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
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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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