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Method for promoting high-efficiency accumulation of arachidonic acid grease

A technology of arachidonic acid and oil, applied in the field of bioengineering, can solve the problems of low production intensity, long production cycle and limited effect of arachidonic acid, and achieve the effects of low cost, short production cycle and simple operation

Active Publication Date: 2012-07-18
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these methods have played a certain regulatory role in the fermentation and production process of arachidonic acid oil, the effect is still very limited, and the problems of low production intensity of arachidonic acid under conditions such as nitrogen limitation, change of dissolved oxygen, and addition of substances are still there. It has been well solved. Although the production intensity of arachidonic acid is improved in the aging process, the production cycle is too long to be about 276h, and these methods are usually not easy to apply in industrialization

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  • Method for promoting high-efficiency accumulation of arachidonic acid grease
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  • Method for promoting high-efficiency accumulation of arachidonic acid grease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: strain activation.

[0028] Strain activation: seeds were cultured on a PDA slant medium at 25° C. for 168 hours, and the slant medium was made of the following materials: 200 g of potatoes, 20 g of glucose, 20 g of agar, and 1000 mL of water.

[0029] Seed activation: the mycelium on the slant is inserted into a 250mL seed shaker flask from the slant to carry out seed activation culture, the liquid volume of the shaker flask is 20% (v / v), and the seed activation medium formula is: glucose 30g / L, yeast Paste 6g / L, KH 2 PO 4 3g / L, NaNO 3 3g / L, MgSO 4 ·7H 2 O 0.5g / L, the solvent is water; pH 6.0, the culture conditions are: temperature 25°C, rotation speed 125rpm, culture time 20h.

Embodiment 2

[0030] Example 2: The rapid proliferation of bacteria in the first stage.

[0031] The seed after the activation of Example 1 is received with the inoculum size of 10% (v / v) and is equipped with 7000mL to be equipped with the fermentation of the carbon source that glucose concentration is respectively 40,60,80 and 100g / L and other nutrient-rich medium Cultivated in tanks, the liquid volume of the fermentation tank is 70% (v / v), and the carbon source and other nutrient-rich medium. The formula of nutrients except carbon source is: yeast extract 11g / L, KH 2 PO 4 3.8g / L, NaNO 3 3.4g / L, MgSO 4 ·7H 2 O 0.5g / L, calcium pantothenate 0.5g / L, vitamin 0.2g / L, solvent is water. The culture conditions are temperature of 25° C., rotation speed of 125 rpm, and the culture time is subject to the consumption of sugar as much as possible. The experimental results are shown in Table 1. It can be seen that when the initial sugar is 60g / L, the glucose is exhausted at 60h, and the productio...

Embodiment 3

[0034] Example 3: The second stage of rapid oil accumulation culture.

[0035] The thalline of embodiment 2 (60g / L glucose condition) is collected, and the thalline collection method is that fermented liquid is centrifuged at 3000rpm for 3min, washed with sterile physiological saline, centrifuged under the same conditions after washing, and the washing and centrifuging operation is repeated twice , to collect bacteria.

[0036] The collected bacteria were cultivated in culture medium with initial glucose concentrations of 20, 40, 60, 80, 100, and 120 g / L, calcium pantothenate 0.25 g / L, vitamin 0.1 g / L, and water as the solvent. For: the temperature is 25°C, the rotation speed is 125rpm, and the incubation time is 48 hours, the experimental results show that the oil accumulation is as follows: figure 2 shown. It can be seen that when the initial sugar is lower than 60g / L, the total oil increases with the increase of the initial sugar concentration, but the residual sugar is ...

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Abstract

The invention discloses a method for promoting the high-efficiency accumulation of arachidonic acid grease. After the mortierella alpina strain is cultured to be activated, rapid mycelium multiplication culture is carried out on culture medium rich in the carbon source and other nutrient substances, mycelia are collected, rapid grease accumulation culture is then carried out on culture medium rich in the carbon source but lack in other nutrient substances, the mycelia are collected, rapid arachidonic acid accumulation culture is carried out on culture medium lacking carbon in the end, and the mycelia are collected. The technique can accurately control each stage of the arachidonic acid grease fermentation process, has the characteristics of short production period, high arachidonic acid production intensity, good grease quality and the like, and is easy to operate and industrially apply.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for promoting efficient accumulation of arachidonic acid oil based on staged changes of medium components. Background technique [0002] Arachidonic acid (ARA, namely 5, 8, 11, 14-eicosatetraenoic acid) belongs to the n-6 series of long-chain polyunsaturated fatty acids (PUFAs), and is the most abundant and widely distributed in the human body. and one of the most active essential PUFAs (Lombardo Y B, Hein G, Chicco A. Lipids, 2007, 42(5):427-437). It is the direct precursor of derivatives such as prostaglandins, cycloprostaglandins and leukotrienes. It has a variety of biological activities and plays an important role in promoting intellectual development, improving human vision, reducing blood lipids, enhancing immunity and anti-cancer. It has a wide range of applications in the fields of biomedicine, cosmetics, functional food and health products. [0003] ...

Claims

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Application Information

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IPC IPC(8): C12P7/64C12R1/645
Inventor 黄和纪晓俊邓中涛聂志奎彭超丛蕾蕾黎志勇
Owner NANJING UNIV OF TECH
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