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Method for producing highly unsaturated fatty acids and lipid containing same

a technology of unsaturated fatty acids and lipids, applied in the field of microorganisms, can solve the problems of low resistance to a high glucose concentration, complicated production steps, and required complicated operations, and achieve the effect of efficient production

Inactive Publication Date: 2006-01-19
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a method for efficiently producing arachidonic acid, dihomo-γ-linolenic acid, and eicosapentaenoic acid or lipid containing these acids using a microorganism belonging to the genus Mortierella. The microorganism has resistance to a carbon source of high concentration, which allows it to grow in a medium containing a high concentration of carbon source at the start of culturing. The microorganism shows a high growth level without using complicated operations such as pH control and addition of inorganic salts, and produces with high productivity the desired acids. The invention also provides animal feed containing the microorganism. The new microorganism has properties that make it a more efficient producer of the desired acids than previously known microorganisms.

Problems solved by technology

However, the microorganisms belonging to the genus Mortierella which have been used in the methods each have a low resistance to a high glucose concentration.
Accordingly, the methods have the problem that the production steps become complicated.
However, in order to suppress the inhibition of the growth of the microorganism caused by the high glucose concentration, procedures such as pH control and addition of salts were fully utilized, and complicated operations were required.
On the other hand, however, increasing the concentration of the carbon source produces a harsh condition for growing the microorganism due to the resultant osmotic pressure, and results in suppressing the growth thereof.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Evaluation of Resistance to Carbon Source

[0071] The following media (pH 6.3) each in an amount of 10 ml were placed in 50-ml Erlenmeyer flasks, respectively: (1) a medium containing 1% glucose and 0.5% yeast extract; (2) a medium containing 2% glucose and 10% yeast extract; (3) a medium containing 3% glucose and 1.5% yeast extract; (4) a medium containing 4% glucose and 2% yeast extract; (5) a medium containing 5% glucose and 2.5% yeast extract; and (6) a medium containing 8% glucose and 1.6% yeast extract. Each of the media was sterilized at 120° C. for 20 minutes.

[0072] The genus Mortierella, strain SAM 2197 (FERM BP-6261) was inoculated into a Czapek agar medium (0.2% NaNO3, 0.1% K2HPO4, 0.05% MgSO4.H2O, 0.05% KCl, 0.001% FeSO4.7H2O, 3% sucrose and 2% agar, pH 6.0) slanted to give a spore formation slant. Eight milliliters of sterilized water was placed therein, and the mixture was agitated, followed by inoculating 100 μl of the spore solution into each of the media.

[0073] The...

example 2

Evaluation of Amount of Arachidonic Acid Produced with Initial Glucose Concentration Increased

[0075] The following media (pH 6.3) each in an amount of 5 liters were placed in 10-liter jar fermenters, respectively: (1) a medium containing 6% glucose, 2% yeast extract, 0.2% soybean oil and 0.01% Adekanol; (2) a medium containing 8% glucose, 2% yeast extract, 0.2% soybean oil and 0.01% Adekanol; and (3) a medium containing 11% glucose, 2% yeast extract, 0.2% soybean oil and 0.01% Adekanol. Each of the media was sterilized at 120° C. for 30 minutes. One hundred milliliters of a preculture solution of the genus Mortierella, strain SAM 2197 (FERM BP-6261) was inoculated into each of the media, and culturing with aeration and agitation was conducted for 8 days at 28° C. at an aeration rate of 1 vvm with stirring at 300 rpm.

[0076] To each of the media was added 2.0% of glucose on the second, the third and the fourth day of culturing. After completion of culturing, the following amounts, p...

example 3

Control for Examples 4 and 5

[0078] One hundred milliliters of a medium (pH 6.0) containing 6% glucose and 2% yeast extract was placed in a 500-ml Erlenmeyer flask, and sterilized at 120° C. for 20 minutes. One hundred microliters the spore solution of the genus Mortierella, strain SAM 2197 (FERM BP-6261) used in Example 1 was inoculated into the medium, and shaking culture was conducted at 28° C. for 8 days using a reciprocal shaker (110 rpm). After culturing, the microbial cells were recovered by filtering, washed with water sufficiently, and freeze dried to give 3,120 mg of dried microbial cells. Fat and oil was extracted from the microbial cells by Bligh & Dyer's extraction using a chloroform-methanol-water single phase system solvent. The amount was 624 mg.

[0079] Methyl esterification of the fat and oil was carried out by treating it with a mixture of anhydrous methanol-hydrochloric acid (10%) at 50° C. for 3 hours, and the esterified products were extracted with ether. The fa...

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Abstract

A method for producing highly unsaturated fatty acids comprising culturing a microorganism, belonging to the genus Mortierella and having resistance to a carbon source, in a medium having a carbon source concentration of at least 4% by weight, and collecting highly unsaturated fatty acids from the cultured products. Culturing the microorganism for about a week gives at least about 7 g / L of highly unsaturated fatty acids.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a microorganism, belonging to the genus Mortierella and having resistance to a carbon source of high concentration, a method for the production of arachidonic acid, dihomo-γ-linolenic acid and / or eicosapentaenoic acid, and of a lipid containing the acids by fermentation using the microorganism, as well as animal feed containing the microorganism. BACKGROUND ART [0002] Attention has been paid to arachidonic acid (5,8,11,14-eicosatetraenoic acid), dihomo-γ-linolenic acid (8,11,14-eicosatrienoic acid) and eicosapentaenoic acid (5,8,11,14,17-eicosapentaenoic acid) because these acids can become precursors of eicosanoids such as prostaglandin, leukotriene and tromboxane, and because the acids themselves have physiological activities. For example, eicosapentaenoic acid has been marketed as a food and medicine on the basis of its antithrombotic preventive action or lipid-lowering action. [0003] Furthermore, it has recently been...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P7/64C12N1/18C12P31/00C12P7/6432C12P7/6427C12P7/6472
CPCA23K1/008C12P7/6472C12P7/6427A23K1/164A23K10/16A23K20/158C12P7/6432
Inventor SUZUKI, OSAMUONO, KAZUHISASHIGETA, SEIKOAKI, TSUNEHIROAKIMOTO, KENGO
Owner SUNTORY HLDG LTD
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