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Low-chlorine medium used for culturing Schizochytrium limacinum and production method for docosahexenoic acid (DHA) by using same

A technology for Schizochytrium and culture medium, which is applied in the field of low-chlorine medium for culturing Schizochytrium, can solve problems such as being expensive and difficult to realize, and achieves the advantages of prolonging service life, reducing chloride ion concentration and reducing production cost. Effect

Active Publication Date: 2014-03-12
青岛海智源生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While there are other grades of stainless steel that are more resistant to chlorine corrosion, they are very expensive and are usually only used in fermentation equipment that produces very valuable compounds
[0006] In theory, reducing the concentration of chloride ions in the medium can minimize the corrosion of equipment such as stainless steel fermenters or other storage tanks that contact fermentation liquid, but in practice it is a process that is difficult to achieve

Method used

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  • Low-chlorine medium used for culturing Schizochytrium limacinum and production method for docosahexenoic acid (DHA) by using same
  • Low-chlorine medium used for culturing Schizochytrium limacinum and production method for docosahexenoic acid (DHA) by using same
  • Low-chlorine medium used for culturing Schizochytrium limacinum and production method for docosahexenoic acid (DHA) by using same

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to marine microorganism cultivation technology, specifically to a medium used for culturing Schizochytrium limacinum and a production method for DHA by using the medium. The medium provided by the invention is a medium with a low chlorine ion concentration and has a chlorine ion concentration of 2 g / L, a potassium ion concentration of higher than 0.1 g / L and a sodium ion concentration of 1 to 20 g / L. With the medium provided by the invention, the output of DHA can basically maintain same or be higher, the degrees of corrosion of the medium on a series of production equipment like fermentation equipment, oil pressing equipment and extraction equipment are reduced, the service life of the equipment is prolonged, production safety is guaranteed, and the chlorine ion concentration in discharged sewage during production is reduced, thereby further reducing production cost.

Description

technical field [0001] The invention relates to the technical field of marine microorganism culture, in particular to a low-chloride medium for cultivating Schizochytrium and a method for producing DHA with the medium. technical background [0002] In recent years, many studies have fully confirmed that long-chain omega-3 fatty acids have beneficial effects on human health. The beneficial effects include lowering blood triglyceride and cholesterol levels, lowering blood pressure, preventing cardiovascular diseases, and inhibiting platelet aggregation. , prevent thrombosis and stroke, prevent inflammation and asthma, lower blood sugar, prevent diabetes, prevent breast cancer and colorectal cancer, promote brain cell development, improve brain cell vitality, improve memory, prevent vision loss, and prevent senile dementia. Several heterotrophic marine microorganisms, including those of the family Thraustochytriaceae, have been found to be able to produce high levels of these i...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 白长军宁超美刘冬英王兴国金青哲初跃峰戚桂斌
Owner 青岛海智源生命科技有限公司
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