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Liquid fermentation production method of docosahexenoic acid (DHA) through solid material cultivation of schizochytrium sp.

A technology of Schizochytrium and liquid fermentation, applied in the field of fermentation and production of DHA, which can solve the problems of long cultivation period, many fermentation stages, and high investment costs, and achieve the effects of simplifying the production process, simplifying the expansion process, and reducing investment

Active Publication Date: 2013-06-12
XIAMEN KINGDOMWAY GROUP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional seed cultivation and fermentation methods involve many stages of fermentation, complicated process, long cultivation period and high investment cost.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Schizochytrium sp. KDW-12 strains were inoculated on slant medium for activation at 25°C for 2 days. The formula of the slant medium is (g / L): glucose 20, peptone 15, yeast powder 3, sea crystal 15, agar powder 20, pH value 6.5, sterilized at 121°C for 30 minutes. Add 0.85% by mass of normal saline to the slant at the end of the culture period to wash the bacteria to obtain a bacterial suspension, and control the concentration of the bacteria to 2.5×10 6 individual / mL.

[0040] The obtained bacterial suspension was directly inoculated on the sterilized solid medium at an inoculation amount of 1% by mass fraction, and cultured at 25°C for 1 day to obtain first-grade solid seeds.

[0041] Under aseptic conditions, the inoculum size of 10% by mass fraction is used to insert the first-level solid material seeds in the solid medium (the first-level solid material seeds used for inoculation are based on 1: 1 (W / V) ratio to wash the bacterial cells to make a bacterial suspen...

Embodiment 2

[0048] Schizochytrium sp. KDW-12 strains were inoculated on slant medium for activation at 28°C for 2 days. The formula of the slant medium is (g / L): glucose 25, peptone 12, yeast powder 3, sea crystal 16, agar powder 20, pH value 6.0, sterilized at 121°C for 30 minutes. Add 0.85% by mass of normal saline to the slant at the end of the culture period to wash the bacteria to obtain a bacterial suspension, and control the concentration of the bacteria to 1.0×10 6 individual / mL.

[0049] The obtained bacterial suspension was directly inoculated on the shake flask liquid medium, the inoculum amount was 1%, the shake flask rotated at 220 rpm, and cultured at 28° C. for 1 day to obtain first-grade liquid seeds. Shake flask liquid seed medium composition (g / L): glucose 25, sodium chloride 20, potassium dihydrogen phosphate 0.2, yeast powder 5, magnesium sulfate heptahydrate 10, sea salt 0.4, glutamic acid 0.4, pH 6.0, 121 Sterilize at ℃ for 30min.

[0050] Under sterile conditions...

Embodiment 3

[0057] Schizochytrium sp. KDW-12 was inoculated on the slant medium for activation at 28°C for 3 days. The formula of the slant medium is (g / L): glucose 30, peptone 12, yeast powder 3, sea crystal 15, agar powder 20, pH value 6.5, sterilized at 121°C for 30 minutes. Add 0.85% by mass of normal saline to the slant at the end of the culture period to wash the bacteria to obtain a bacterial suspension, and control the concentration of the bacteria to 2.0×10 6 individual / mL.

[0058] The obtained bacterial suspension was directly inoculated on the liquid culture medium of the shake flask according to the inoculation amount of 1% by mass fraction, the rotation speed of the shake flask was 220rpm, and cultured at 28°C for 1 day to obtain the first-grade liquid seeds. Shake flask liquid seed medium composition (g / L): glucose 20, sodium chloride 22, potassium dihydrogen phosphate 0.2, yeast powder 3, magnesium sulfate heptahydrate 10, sea salt 0.3, glutamic acid 0.6, pH 6.5, 121 Ste...

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Abstract

The invention relates to a fermentation production method of docosahexenoic acid (DHA), in particular to a liquid fermentation production method of docosahexenoic acid (DHA) through solid material cultivation of the schizochytrium sp. KDW-12. An applied bacterium strain is the schizochytrium sp. KDW-12. The liquid fermentation production method of the DHA includes the following steps of activation of the bacterium strain, preparation of bacterium suspension, preparation of one-level seed, preparation of two-level solid material seed, enlarging cultivation of the two-level solid material seed in a fermentation tank, collection of thallus cells after fermentation, and extraction of DHA oil. The extracted DHA can be used for a nutrient supplement and provides long-chain polyunsaturated fatty acid needed by human and animals. A seed preparation method is improved, the seed enlarging technology is simplified, a seed culture and fermentation period is shortened, and therefore investment is reduced and cost is saved. In addition, a solid material culture medium is full of nutriments and is rich in a plurality of growth factors and more beneficial to growth of the seed. Bacterium strain cells are strong in viability and good in synchronism. Besides, waste water, waste residue and environmental pollution which are generated in the stage of solid material culture are less.

Description

technical field [0001] The invention relates to a method for producing DHA by fermentation, in particular to a method for producing DHA by solid material culture Schizochytrium liquid fermentation. Background technique [0002] Docosahexanoic acid (22:6△4,7,10,13,16,19, referred to as DHA), commonly known as "brain gold", belongs to omega-3 polyunsaturated fatty acids, has the function of nourishing the brain Brain, prevent senile dementia; improve eyesight, prevent and treat myopia; prevent high blood pressure, arteriosclerosis, arthritis and treat cancer and other physiological functions. Therefore, DHA has a broad market prospect as a new generation of functional health care factor. With the expansion of market demand, the requirements for the quality of DHA are getting higher and higher, and the research on DHA production is gradually developing towards the synthesis of biotechnology. The DHA from traditional fish oil has been gradually replaced by DHA from algae oil. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/64C12R1/645C12P7/6434
CPCC12R2001/89C12N1/125C12P7/6434
Inventor 陈金卿陈芳芳陈俊煌林超吴美琼
Owner XIAMEN KINGDOMWAY GROUP
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