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Schizochytrium limacinum and method or fermenting and producing DHA (Docosahexaenoic Acid) grease utilizing high density of schizochytrium limacinum

A technology of high-density fermentation and Schizochytrium, which is applied in the application field of marine microorganisms, can solve the problems of excessive fishing, reduced content, difficult purification process, etc., and achieves the effect of improving price advantages.

Active Publication Date: 2013-01-23
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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AI Technical Summary

Problems solved by technology

[0003] The traditional raw material for the production of DHA is mainly fish oil, but the production of DHA through fish oil has the following disadvantages: (1) fish oil resources are limited, the output is unstable, and the output and quality of fish oil fluctuate greatly, which is far from meeting the market demand
(2) The DHA content in fish oil is not high, accounting for only 7% to 14%, and it is difficult to separate from a large amount of EPA and other highly unsaturated fatty acids with similar structures (3) The purification process is complicated, the production cost is high, and the product yield Low
In the actual production process, omega-3 polyunsaturated fatty acids are hydrogenated and saturated, reducing their content in fish oil, resulting in waste of raw materials, and also impairing the quality of DHA and EPA (4) Fish oil is easy to oxidize and is difficult to apply Food additive industry
Because fish oil contains a heavy and unpleasant fishy smell, it is difficult to remove it even after complex purification processes, which limits the application range of this type of DHA
(5) The increasing market demand based on DHA and other omega-3 polyunsaturated fatty acids will lead to the emergence of overfishing behavior, which is not conducive to the protection of environmental resources
Among them, patents CN00135338.1, CN200410075426.X, CN200610028869.2 and CN 200910061419.7 adopted simple culture methods and optimization of culture conditions, resulting in low biomass, the highest being only 42.5g / L, and no real high density fermentation
Patent CN200910033869.5 achieves a higher fermentation density, and the yield reaches 70g / L, but the oil content accumulated in the cells is not high, only 31.5g / L

Method used

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  • Schizochytrium limacinum and method or fermenting and producing DHA (Docosahexaenoic Acid) grease utilizing high density of schizochytrium limacinum
  • Schizochytrium limacinum and method or fermenting and producing DHA (Docosahexaenoic Acid) grease utilizing high density of schizochytrium limacinum
  • Schizochytrium limacinum and method or fermenting and producing DHA (Docosahexaenoic Acid) grease utilizing high density of schizochytrium limacinum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Effects of Different Carbon Sources and Carbon Source Concentrations on the Growth and Oil Accumulation of Schizochytrium SD116 Strain

[0032] In a 250ml Erlenmeyer shaker flask, configure 50ml of medium, the nitrogen source is yeast extract 20g / L, the salinity is 15, add different carbon sources (glycerol, glucose, fructose, xylose, sucrose, maltose and starch) , the concentration is 60g / L, the pH value is adjusted to 6.0, after autoclaving, insert 5ml of the pre-cultivated strain seed solution, and vibrate at 25°C for 5 days on an air bath shaker, and the shaking speed is 180rpm. The bacteria were collected by centrifugation, freeze-dried to constant weight, and their dry weight was measured; some of the bacteria were taken, and the oil was extracted and methylated by the conventional chloroform-methanol fermentation method, and the percentage content of DHA in the bacteria was determined by GC-MS. The results are shown in Table 1

[0033]After determinin...

Embodiment 2

[0041] Example 2: Effects of Different Nitrogen Sources and Nitrogen Source Concentrations on the Growth and Oil Accumulation of Schizochytrium SD116 Strain

[0042] In a 250ml Erlenmeyer shaker flask, configure a 50m culture medium, use glucose as a carbon source, a concentration of 60g / L, and a salinity of 15, add organic nitrogen sources (yeast extract, peptone and tryptone) with a concentration of 20g / L respectively and 5g / L inorganic nitrogen source (urea, ammonium acetate and sodium nitrate), adjust the pH value to 6.0, after autoclaving, insert 5ml of pre-cultivated strain seed liquid, and vibrate on an air bath shaker at 25°C For 5 days, the shaking speed was 180 rpm. The analysis method is the same as above, and the experimental results are shown in Table 3.

[0043] Table 3 Effects of different nitrogen sources on the growth and oil accumulation of Schizochytrium SD116 strain

[0044]

[0045] It can be seen from Table 3 that yeast extract and peptone can better...

Embodiment 3

[0050] Example 3: Effects of Different Temperatures and Salinity on Growth and Oil Accumulation of Schizochytrium SD116 Strain

[0051] Schizochytrium growth and oil accumulation are affected by temperature and salinity. The experiment adopts optimized carbon source, nitrogen source and concentration, and designs different temperatures (20-37° C.) and salinity (0-60). The experimental method and analysis method are the same as the above examples.

[0052] Table 5 Effects of different temperatures on the growth and oil accumulation of Schizochytrium SD116 strain

[0053]

[0054]

[0055] It can be seen from Table 5 that the lower the temperature, the higher the DHA content of the Schizochytrium SD116 strain, indicating that low temperature is conducive to the accumulation of DHA; when the temperature is 25-28 °C, the growth of the SD116 strain is the largest, reaching 29.56 g / L, but when the temperature was higher than 30℃, the biomass and oil content of Schizochytrium ...

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Abstract

The invention belongs to the field of microbial fermentation engineering, and discloses schizochytrium limacinum (Aurantiochytrium sp.SD116). The schizochytrium limacinum is stored in China Microbial Culture Collection Administration Committee General Microbial Center with a number of CGMCC No: 6208), and a method for fermenting and producing DHA (Docosahexaenoic Acid) grease utilizing high density of the schizochytrium limacinum. According to the schizochytrium limacinum and the method disclosed by the invention, a bacterial strain fermenting condition is optimized based on element supply and a fermenting control angle, a carbohydrate supplementation operation is carried out, therefore, high-density fermentation is achieved, dry cell weight finally reaches 70.43g / L, grease content reaches 50.1g / L, DHA occupies more than 35% of total content of fatty acid; moreover, bioactive substances such as beta-carotene, astaxanthin and squalene are contained. The complete set of technology is convenient to operate; higher biomass liveweight and DHA content can be obtained; fermenting cost can be reduced; and the method is suitable for industrial fermenting production.

Description

technical field [0001] The invention relates to the application technology of marine microorganisms, in particular to a marine fungus Schizochytrium SD116 strain and a method for producing DHA by high-density fermentation thereof. Background technique [0002] Docosahexaenoic acid (DHA) is an important omega-3 polyunsaturated fatty acid, which has important physiological functions such as promoting brain cell development, lowering blood fat, protecting eyesight, anti-cancer and improving immunity, and is widely used in infants. Children's food and pharmaceutical industry. In addition, DHA is also an essential fatty acid required for the growth and development of various seawater fish, which can improve the survival rate of fry and reduce the incidence of albinism. [0003] The traditional raw material for the production of DHA is mainly fish oil, but the production of DHA through fish oil has the following disadvantages: (1) fish oil resources are limited, the output is uns...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 崔球高莽宋晓金冯银刚
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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