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Schizochytrium sp. and method for producing DHA lipa by using same

A technology of schizovibrio and oil, which is applied in the field of marine fungus schizovibrio and its high-density fermentation to produce DHA oil, can solve the problems of low biomass, low production efficiency, increase production cost and the like, and achieve biomass and high DHA content, lower fermentation cost and simple process

Active Publication Date: 2009-11-11
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, there are four papers related to the present invention. The bacteria used in CN200610125476.3 is Cryptodinosaurus, with a biomass of 20-40g / L and an oil content of 20-50% in the cells; CN00135338.1, CN200410075426.X , CN200610028869.2 all adopt Schizovibrio bacterium as the production bacteria, the process and fermentation method are more traditional, only the optimization of simple culture method and culture conditions, there is no detailed study on the key factors that directly affect the biomass accumulation, resulting in the biomass is not High, up to 42.5g / L, did not achieve high-density fermentation in the true sense
Low production efficiency increases production costs, making it difficult for DHA derived from microorganisms to have a significant price advantage over DHA derived from fish oils

Method used

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  • Schizochytrium sp. and method for producing DHA lipa by using same
  • Schizochytrium sp. and method for producing DHA lipa by using same
  • Schizochytrium sp. and method for producing DHA lipa by using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Investigation on the basic growth performance of bacterial strains.

[0042] The basic properties of the screened strain CCTCC No: M 209059 were investigated, including culture in different liquid media (see Table 1) and culture in different solid media (see Table 2).

[0043] Table 1 Culture morphology and fermentation results of Schizovibrio in different liquid media

[0044]

[0045] Remarks: Culture medium 1-4 is cultured with natural seawater, the salt concentration is (15, 30, 45, 60g / L), and the medium also includes 40g / L glucose and 4g / L yeast extract. Medium 5 is self-prepared artificial seawater medium, including glucose 40g / L, yeast extract 3g / L, KH 2 PO 4 4g / L, NaCl 15g / L, MgSO 4 ·7H 2 O 5g / L, KCl 2g / L, MgCl 2 3g / L, FeCl 3 .6H 2 O 0.1g / L, sodium glutamate 10g / L. The total osmotic pressure is 1092mOsm L -1 .

[0046] Table 2 Culture characteristics of Schizovibrio on different solid media

[0047]

[0048] Remarks: PDA, GYP, LB, ...

Embodiment 2

[0050] Example 2: Study on osmotic pressure tolerance of Schizovibrio bacterium.

[0051] Schizovibrio is a kind of marine fungus, which has a stronger ability to resist osmotic pressure than other microorganisms. In this embodiment, on the basis of the initial sugar concentration of 100g / L, the concentration of self-prepared artificial seawater was changed, and the osmotic pressure was selected to be 1208.56, 1715.46, 2729.26, 3743.06, 4756.86mOsm L -1 Five gradient experiments.

[0052] There are two main forms of Schizovibrio in liquid culture, swimming cells and static cysts. In the early stage, in order to reproduce, the flagella degenerate and are encapsulated, and the cells divide and reproduce in the cyst; in the later stage, the cyst ruptures, the cells are released, and the cells are in two states of swimming or resting, and the swimming cells have flagella. In the first two gradients, the division of Schizovibrio was vigorous, and the cell growth was fast, and the...

Embodiment 3

[0055] Example 3: Effects of supply of nitrogen, phosphorus and sulfur on the biomass of Schizovibrio and the content of DHA oil.

[0056] In the 250ml fluted flask, keep the optimum osmotic pressure (1715.46mOsm L -1 ) conditions, optimize the supply of nitrogen, phosphorus, sulfur and other elements, and investigate their effects on biomass and oil content. Yeast extract and ammonium salt were selected as the nitrogen source, the yeast extract concentration was kept constant, the ammonium salt concentration was changed, and the final nitrogen concentration was selected as five gradient experiments of 0.8742, 1.2985, 1.7227, 2.1470, and 2.517g / L, and their effects on biomass and fatty acids were compared. The results are shown in Table 4.

[0057] Table 4 Comparison of biomass and fatty acid content at different nitrogen concentrations

[0058]

[0059] Nitrogen source concentration was closely related to fatty acid accumulation. Sufficient nitrogen source can ensure ra...

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Abstract

The invention discloses a Schizochytrium sp., which has a category name of Schizochytrium sp., is preserved in Common Microorganisms Center of China Committee for Culture Collection of Microorganisms and has a preservation number of CCTCC No.209059. The invention also discloses a method for producing DHA lipa by using the Schizochytrium sp. The method optimizes a strain fermentation medium from the perspectives of osmotic pressure and element supply and combines a fed-batch strategy to achieve high-density fermentation of microalgae, thus the final dry cell weight reaches 70 grams per liter, the lipa content reaches 31.5 grams per liter, and the DHA accounts for more than 35 percent of the total fatty acid content. All indexes of the DHA essential oil obtained by the method accord with the standard of food additives, and the DHA essential oil contains a bioactive substance of squalene. The method has the advantages of simple entire process, convenient operation, and high biomass and DHA content, reduces the fermentation cost, and is suitable for industrial production.

Description

technical field [0001] The invention relates to a marine fungus schizovibrio and a method for producing DHA oil by high-density fermentation thereof, belonging to the field of biotechnology. Background technique [0002] Docosahexaenoic acid (DHA) is an important omega-3 long-chain polyunsaturated fatty acid, which has important physiological functions such as promoting brain cell development, lowering blood lipids, protecting eyesight, anti-cancer, and improving immunity. It is widely used In infant food and pharmaceutical industry. [0003] Traditional DHA production raw materials mainly come from fish oil, but differences in fish species, seasons, geographical locations and other factors have caused instability in oil content and unsaturated fatty acid content in oil; DHA extracted from fish oil has high cholesterol content and contains a large amount of EPA (Eicosapetaenoic acid, 22:5) and various minerals; coupled with the increasingly scarce source of fish oil, it is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/64C12R1/01
Inventor 黄和任路静肖爱华金立晶李娟
Owner NANJING UNIV OF TECH
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