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Method for producing arachidonic acid oil by Mortierella alpina fermentation on basis of dissolved oxygen control

A technology of Mortierella alpine and arachidonic acid, which is applied in the field of microbial fermentation, can solve problems such as rise, and achieve the effects of simple process operation, improved fermentation capacity, and good profit.

Active Publication Date: 2015-01-14
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the currently reported experiments with high net content of arachidonic acid are basically carried out at the level of 250mL shake flasks, 1L, 5L, and 20L small fermentation tanks, and there are few good examples reported in industrial scale-up
[0006] Therefore, in view of the disadvantages of the existing mechanically stirred fermenters and airlift fermenters on the influence of the bacterial morphology, for the fermentation of Mortierella alpina to produce arachidonic acid oil, the cycle is long, extremely aerobic, and the viscosity increases significantly in the later stage of fermentation. The fermentation system optimizes the appropriate high-density fermentation process conditions, and provides a new process suitable for the accumulation of Mortierella alpine bacteria to produce arachidonic acid oil, which increases the biomass of the bacteria and the content of arachidonic acid oil, and improves the quality of peanuts. Tetraenoic acid production intensity, and it is necessary to put it into industrialized large-scale production

Method used

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  • Method for producing arachidonic acid oil by Mortierella alpina fermentation on basis of dissolved oxygen control
  • Method for producing arachidonic acid oil by Mortierella alpina fermentation on basis of dissolved oxygen control
  • Method for producing arachidonic acid oil by Mortierella alpina fermentation on basis of dissolved oxygen control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Fermentation of Mortierella alpina in a 7.5L tank to produce arachidonic acid oil based on dissolved oxygen control strategy

[0031] 1. Activation of strains and preparation of seed solution: Mortierella alpina strain R807 (CCTCC M 2012118) was selected as the starting strain, and the preserved strains were inserted into PDA slant medium, and placed in an incubator at 25°C Cultivate for 10 days, transfer to a 500mL grooved bottle containing 100mL seed medium and culture in a constant temperature shaker, the inoculum size is 10% (v / v, 10mL), the culture conditions are 120rpm, the temperature is 25°C, and the culture time is 1 -2 days.

[0032] 2. Inoculate the seed liquid obtained in step 1 into a 7.5L tank (New Brunswick Scientific, USA), the liquid volume is 5L, the inoculum volume is 10% (v / v, 500mL), and the culture temperature is controlled at 28°C for 0-120h , 121-216h controlled at 18°C,

[0033]At the beginning of fermentation, the seed li...

Embodiment 2

[0041] Example 2. Arachidonic acid produced by fermenting Mortierella alpina in a 7m3 tank based on the dissolved oxygen regulation strategy

[0042] 1. Activation of strains and preparation of seed solution: Mortierella alpina strain R807 (CCTCC M 2012118) was selected as the starting strain, and the preserved strains were inserted into PDA slant medium, and placed in an incubator at 25°C Cultivate for 10 days, transfer to a 500mL grooved bottle containing 100mL seed medium and culture in a constant temperature shaker, the inoculum size is 10% (v / v, 10mL), the culture conditions are 120rpm, the temperature is 25°C, and the culture time is 1 -2 days, prepare a first-grade seed solution. Then, according to the inoculation amount of 10% (v / v), the primary seed solution was inserted into a 100L seed tank, the cultivation conditions were 200 rpm, the temperature was 25°C, and the cultivation time was 1 day to prepare the secondary seed solution. Next, all the prepared seco...

Embodiment 3

[0051] Example 3. Arachidonic acid produced by fermenting Mortierella alpina in a 25m3 tank based on the dissolved oxygen regulation strategy

[0052] 1. Activation of strains and preparation of seed solution: Mortierella alpina strain R807 (CCTCC M 2012118) was selected as the starting strain, and the preserved strains were inserted into PDA slant medium, and placed in an incubator at 25°C Cultivate for 10 days, transfer to a 500mL grooved bottle containing 100mL seed medium and culture in a constant temperature shaker, the inoculum size is 10% (v / v, 10mL), the culture conditions are 120rpm, the temperature is 25°C, and the culture time is 1 -2 days, prepare a first-grade seed solution. Then, according to the inoculation amount of 10% (v / v), the primary seed solution was inserted into a 150L seed tank, the cultivation conditions were 200rpm, the temperature was 25°C, and the cultivation time was 1 day to prepare the secondary seed solution. Next, all the prepared sec...

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Abstract

The invention relates to a method for producing arachidonic acid oil by Mortierella alpina fermentation on the basis of dissolved oxygen control. The method comprises the following steps: carrying out progressive amplification culture on the Mortierella alpina strain in a test tube slant culture medium and a shake culture medium to prepare a seed solution, and inoculating in a fermentation tank to perform fermentation, wherein the whole fermentation process is divided into an early fermentation stage, a medium fermentation stage and a later fermentation stage. The initial rotation speed is set at 0 rpm, and the initial dissolved oxygen is set at 100%; in the early fermentation stage, the dissolved oxygen is lowered and stable at 40-70%; in the medium fermentation stage, the dissolved oxygen is controlled at 20-40%; and in the later fermentation stage, the dissolved oxygen is controlled at 40-70%. After the fermentation is finished, the Mortierella alpina cell dry weight, oil content, percent of arachidonic acid in total oil, and unit yield of arachidonic acid can respectively reach 55g / L, 54%, 65% and 20.628g / L, and the arachidonic acid production intensity is up to 2.292g / (L*d).

Description

technical field [0001] The invention relates to a method for producing arachidonic acid oil by regulating Mortierella alpine fermentation based on dissolved oxygen, and belongs to the technical field of microbial fermentation. Background technique [0002] Since the 1990s, people have paid more and more attention to healthy diet and nutrition, and long-chain polyunsaturated fatty acids have gradually come into people's sight. Long-chain polyunsaturated fatty acids mainly include arachidonic acid (ARA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). These fatty acids not only play an important role in the composition of membrane phospholipids, but also can be used as precursors for the synthesis of signal molecules eicosanoids (including prostaglandins, thromboxanes and leukotrienes, etc.), and are widely used in Pharmaceuticals and cosmetics, especially in the field of infant food and nutritional products. [0003] At present, the process of producing ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q3/00C12P7/64C12R1/645
Inventor 纪晓俊邬文嘉张瑷珲黄和
Owner NANJING UNIV OF TECH
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