A low-chloride medium for cultivating Schizochytrium and a method for producing DHA using the medium
A technology of Schizochytrium and culture medium, which is applied in the field of low-chloride medium for cultivating Schizochytrium, can solve the problems of being expensive and difficult to realize, and achieve the goals of prolonging the service life, reducing the concentration of chloride ions, and reducing production costs Effect
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Embodiment 1
[0018] Embodiment 1: the configuration of Schizochytrium low chlorine culture medium
[0019] According to the formulation in the following table 1, the total concentration of chloride ions is the control medium (code-named "0") of 10.35g / L, the low-chloride medium (code-named "1") that the total concentration of chloride ions is 1g / L and Low-chloride medium (code-named "2") with a total concentration of chloride ions of 0.30 g / L. The source of chloride ions in the control culture medium whose code is "0" includes sodium chloride (wherein the concentration of chloride ions is 10.32g / L) and calcium chloride (wherein the concentration of chloride ions is 0.03g / L); The sources of chloride ions in the low-chloride medium of 1" include sodium chloride (in which the concentration of chloride ions is 0.97g / L) and calcium chloride (in which the concentration of chloride ions is 0.03g / L); the culture code-named "2" The sources of chloride ions in the base include sodium chloride (wher...
Embodiment 2
[0024] Embodiment 2: Shake flask culture Schizochytrium (Schizochytrium) produces DHA in low chlorine medium
[0025] 1) Inoculation culture
[0026] Schizochytrium (Schizochytrium) was inoculated into the 250mL Erlenmeyer flask equipped with "0", "1", and "2" aseptic medium in Example 1, the inoculum size was 5%-10%, temperature 28 ℃, rotating speed 200-220r / min, pH=6.5, and culture on a shaker in a constant temperature culture room for 3-5 days.
[0027] 2) Index test (biomass, total fat content and DHA content)
[0028] (1) Biomass test: take a certain volume of dry cells, and test the quality of the cells after freeze-drying;
[0029] (2) Total fat test: Use concentrated HCl acid hydrolysis method to break the cell wall, use a fat analyzer, use anhydrous ether or n-hexane to extract the soluble matter, remove the solvent to get its oil content;
[0030] (3) DHA content test: use the method of gas chromatography to test the content of DHA in oil.
[0031] The test resul...
experiment example 1
[0034] Experimental Example 1: Effect of Amino Acids on the DHA Production Ability of Schizovibrio
[0035] 1) Preparation of culture medium
[0036] Prepare medium No. 1-3 according to the recipe in Table 3 below. The concentration of chloride ions in the medium is 0.3g / L (300ppm), the total concentration of sodium ions is 7.5g / L, and the total concentration of potassium ions is 1.04g / L. The culture medium was autoclaved.
[0037] Table 3 The composition of medium
[0038]
[0039] Note: 1 Sterilize separately at 115°C for 20-30 minutes.
[0040] The 2 components were sterilized by filtration through a 0.2 μm filter; stored in the dark at 4°C. Add aseptically to the sterilized medium.
[0041] 2) Inoculation culture
[0042] Inoculate Schizovibrio bacterium into a 250mL Erlenmeyer flask containing No. 1-3 sterile medium, the inoculum amount is 5%-10%, the temperature is 28°C, the rotation speed is 200-220r / min, pH=6.5, and cultured at a constant temperature Incubate...
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