DHA fermenting high-yielding method

A technology of strains and strong oxidants, applied in the direction of using microorganisms, methods based on microorganisms, fermentation, etc., can solve the problems of blindness in the breeding process of strains, achieve physiological and biochemical stability, high efficiency, sugar and nitrogen consumption rate accelerated effect

Active Publication Date: 2016-02-17
南京脂禾生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the relatively blind technical problems of the strain selection process in the existing methods

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 This embodiment illustrates the source of the bacterial strain and the culture medium used

[0039] The strain is Schizochytrium HX-308, CCTCCNo: M209059, which was isolated and preserved by Nanjing University of Technology.

[0040]Activation medium and seed medium: glucose 20-60g / L, yeast extract 2-4g / L, MgCl 2 3~6g / L, CaCl 2 2H 2 O1~4g / L, KH 2 PO 4 2~4g / L, sodium glutamate 8~12g / L, KCl1~4g / L, NaCl15~20g / L, FeCl 3 0.1~0.3g / L, MgSO 4 ·7H 2 O1~5g / L.

[0041] Wherein, when preparing the specific medium in each embodiment, one concentration can be selected for the test configuration.

[0042] Domesticated seed medium: glucose 20-60g / L, yeast extract 2-4g / L, MgCl 2 3~6g / L, CaCl 2 2H 2 O1~4g / L, KH 2 PO 4 2~4g / L, sodium glutamate 8~12g / L, KCl1~4g / L, NaCl15~20g / L, FeCl 3 0.1~0.3g / L, MgSO 4 ·7H 2 O1~5g / L, different concentrations of hydrogen peroxide or potassium permanganate.

[0043] Wherein, when preparing the specific culture medium in each em...

Embodiment 2

[0046] Embodiment 2 This embodiment illustrates the specific steps of Schizochytrium domestication

[0047] Activation of Schizochytrium with docosahexaenoic acid production ability for one generation, the activation method is as follows:

[0048] Inoculate in the medium containing 0.02% hydrogen peroxide or potassium permanganate, the inoculum amount is 1% (v / v), and cultivate at 25°C for 24 hours. After 3 days of acclimation, when the Schizochytrium adapted to the environment of 0.02% strong oxidant, it was inserted into the normal medium and cultured for one generation. Then the domesticated strains were inserted into the medium containing 0.03% strong oxidant, and domesticated for 5 days. When the Schizochytrium adapted to the environment of 0.03% strong oxidant, they were inserted into the normal medium and cultivated for one generation. Then the acclimatized strains were inserted into the culture medium containing 0.04% strong oxidant, and acclimatized for 5 days. This...

Embodiment 3

[0049] The domesticated bacterial strain of embodiment 3 different domesticated concentrations and the fermentation performance contrast of original bacterial classification

[0050] Every time the concentration of hydrogen peroxide or potassium permanganate was increased, the domesticated strains were fermented and verified to investigate the changes in the ability of the domesticated strains to produce DHA. Insert the above-mentioned seed solution with 10% (v / v) inoculation amount into 100mL fermentation medium sterilized at 121°C for 30min, the fermentation culture conditions are: temperature 25°C, rotation speed 150rpm, 90g / L glucose Fermentation was terminated after depletion, and the dry cell weight, fatty acid content and DHA content were measured respectively. The fermentation parameters were compared with those of the original strain under the same conditions.

[0051] Table 1: Comparison of fermentation results of domesticated strains of hydrogen peroxide and origin...

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Abstract

The invention provides a DHA fermenting high-yielding method. The DHA fermenting high-yielding method includes the steps that schizochytrium limacinum strains are domesticated through a culture medium with a strong oxidant, the domesticated strains are cultured through a fermenting culture medium, and fermenting high yielding of DHA can be achieved accordingly. After the domesticated strains are fermented, the DHA content is 57.9%, and oil-producing efficiency is 0.492 g/L/h; compared with a control group, the DHA content and the oil-producing efficiency are increased about 20% and 55% respectively. The domesticating method does not involve a genetic engineering method, does not harm the environment, does not increase manpower and material resources, is simple and convenient, and has economic benefits.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the domestication of Schizochytrium strains and a method for producing DHA by using the domesticated strains. Background technique [0002] Docosahexaenoic acid (22:6n-3), referred to as DHA, is an important polyunsaturated fatty acid (polyunsaturated fatty acid, PUFA) in the omega-3 series. DHA is not only an essential substance for the normal development of the brain and vision of infants, but also has the functions of delaying the decline of brain and vision functions in the elderly, reducing blood lipids, inhibiting thrombosis, resisting autoimmune reactions, and inhibiting tumors. Therefore, it is known as "brain gold". ". At present, DHA has been widely used in products such as infant food, health food and auxiliary therapeutic agent. [0003] At present, the DHA oil derived from traditional fish oil is easily affected by the environment and seasons, resulting in limited DHA pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/36C12P7/64C12R1/645
Inventor 黄和孙小曼任路静纪晓俊
Owner 南京脂禾生物科技有限公司
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