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Schizochytrium limacinum suitable for high-density culture and method for producing grease rich in DHA

A technology for high-density cultivation of Schizochytrium, which is applied in the field of Schizochytrium and its production of oil rich in DHA, can solve the problems of high production cost, inability to produce on a large scale, large amount of corn steep liquor solution, etc., and achieve high Biomass and DHA production, reduced production costs, and faster growth

Inactive Publication Date: 2016-03-23
TONGWEI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this is only the result of high-density fermentation culture in 3L and 5L fermenters, which cannot be mass-produced
And in its fermenting and cultivating process, it needs to feed 50% (w / v) corn steep liquor solution for a long time, adopted sugar supplement operation in the whole fermentation process, the corn steep liquor solution consumption is large, the production cost is too high, is not suitable for mass production

Method used

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  • Schizochytrium limacinum suitable for high-density culture and method for producing grease rich in DHA
  • Schizochytrium limacinum suitable for high-density culture and method for producing grease rich in DHA
  • Schizochytrium limacinum suitable for high-density culture and method for producing grease rich in DHA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Mutation and screening of Schizochytrium

[0026] 1.1 UV mutagenesis

[0027] The starting strain is (Schizochytrium sp.) SR21 (purchased from ATCC, USA), and the specific steps are:

[0028] (1) Inoculate the preserved bacterial strain (1.0ml thawed bacterial solution) into 50ml seed culture medium, and culture it on a shaking table at 20-30°C and 150-220rpm for 20-24h to obtain the logarithmic phase bacterial solution; then press 5- 10% (V / V) was transferred to fresh seed medium, and cultivated to the logarithmic phase under the same conditions.

[0029] (2) Wash and centrifuge the logarithmic phase bacterial liquid obtained above through sterile distilled water, and make bacterial suspension, the bacterial liquid concentration OD650 of bacterial suspension is 0.3-0.8; Take appropriate amount of bacterial suspension and pour it into a petri dish, make The bacterial liquid becomes a thin layer of bacterial film with a thickness of 1-5mm; irradiate for 20-100s at a...

Embodiment 2

[0045] 20L High Density Fermentation Production of Schizochytrium sp. SLTW3

[0046] Specific steps are as follows:

[0047] (1) Bacterial strain activation culture: culture the bacterial strain stored in the glycerol tube by streaking on the plate medium (glucose 5g / L, yeast powder 1g / L, peptone 1g / L, sea salt 15g / L, agar powder 17g / L, pH is 6.0), after 40h of cultivation, a single colony was obtained.

[0048] (2) Primary seed culture: Insert single colonies in the plate into a medium containing 50 mL of seed medium (glucose 20 g / L, yeast powder 10 g / L, peptone 5 g / L, seawater 15 g / L, pH 6.0) In a 250mL shake flask, cultured on a shaker at 28°C and 200rpm for 20h to obtain first-grade seeds.

[0049] (3) Secondary seed culture: insert the primary seeds into a 500mL shaker containing 100mL seed medium (glucose 20g / L, yeast powder 10g / L, peptone 5g / L, seawater 15g / L, pH 6.0) In the bottle, the inoculum amount was 10% (V / V), and cultured on a shaker at 28° C. and 200 rpm for...

Embodiment 3

[0056] 1000L High Density Fermentation Production of Schizochytrium sp. SLTW3

[0057] When carrying out large-scale high-density cultivation, shake flasks can be used as the seed culture containers of large fermenters, but a large number of shake flasks is also required, and the possibility of bacterial contamination is high. Preferably, multiple fermentation tanks are used for joint culture, which can effectively avoid bacterial contamination. This example is 1000L high-density fermentation production, using 20L-100L-1000L triple fermentation culture, the specific steps are as follows:

[0058] (1) Bacterial strain activation culture: culture the bacterial strain stored in the glycerol tube by streaking on the plate medium (glucose 5g / L, yeast powder 1g / L, peptone 1g / L, sea salt 15g / L, agar powder 17g / L, pH is 6.0), after 40h of cultivation, a single colony was obtained.

[0059] (2) Primary seed culture: Insert single colonies in the plate into a medium containing 50 mL o...

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Abstract

The invention relates to the technical field of microorganisms, in particular to schizochytrium limacinum suitable for high-density culture and a method for producing grease rich in DHA. The schizochytrium limacinum is suitable for high-density culture and can be used for producing grease rich in DHA, the production method is low in cost, and the biomass and DHA yield are high. The classification naming of the schizochytrium limacinum suitable for high-density culture is (Schizochytrium sp.) SLTW3 and is preserved in the China Center for Type Culture Collection with the preservation serial number of CCTCC No: M2015591. According to the method for producing grease rich in DHA by means of the schizochytrium limacinum, the schizochytrium limacinum is used as the original strain, thalli are collected through high-density fermentation, and then grease rich in DHA is extracted. With the schizochytrium limacinum as the original strain, total biomass can reach 170-190 g / L after high-density fermentation culture, and the DHA yield can reach 25 g / L.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to high-density cultivable Schizochytrium and a method for producing oil rich in DHA. Background technique [0002] DHA is an important component of the cell membrane in the human brain and retinal tissue. It is an n-3 series polyunsaturated fatty acid system necessary for the human body's own development. It plays a vital role in the development of the brain and vision of infants. It has significant effects on diseases such as hypertension, arteriosclerosis, arthritis and cancer. In addition, n-3 series unsaturated fatty acids are also necessary for the growth of fish and shrimp, but they cannot be synthesized by themselves, and must be provided by feed during breeding, especially in the young stage. DHA deficiency will lead to increased mortality and body surface albinism and other diseases. The traditional DHA production route mainly comes from the extraction and purification o...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64C12R1/645
CPCC12P7/6409C12P7/6436C12N1/145C12R2001/645
Inventor 曾娟张良罗国强苏艳秋刘海燕廖利民吴晓娟刘梅肖尧
Owner TONGWEI
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