Mortierella isabellina and application thereof

A kind of technology of Mortierella xanthos, classified and named, applied in the field of bioengineering, can solve the problem of low gamma-linolenic acid yield, etc., and achieve the effects of wide growth range, simple nutritional requirements and good development prospects

Inactive Publication Date: 2011-01-19
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are so many strains capable of producing γ-linolenic acid, there have been few reports on the screening of wild high-yielding bacteria in recent years. Most studies have focused on the transformation of existing strains and metabolic regulation. The improvement of γ-linolenic acid is less, and most of the currently published invention patents related to γ-linolenic acid are Cunninghamella echinlata (application numbers: 95111964.8, 95119429.1, 200610065889.7, 91106190.8), rarely involving other strains, delaying The industrialization process of gamma-linolenic acid has been confirmed, so the screening of microbial resources with high yield of gamma-linolenic acid is conducive to expanding the research space, and at the same time, it can also meet the needs of gamma-linolenic acid industrialization and promote the industrialization process, which is of great significance

Method used

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  • Mortierella isabellina and application thereof
  • Mortierella isabellina and application thereof
  • Mortierella isabellina and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Screening method for high-production GLA strains.

[0034] On December 24, 2009, soil samples were collected from the surface of the tea garden of Nanjing University of Technology, weighed 3.0 grams, baked at 90 ° C for 36 hours, added to 100 mL of sterile water to make a suspension, and then diluted with sterile water to 10 -6 , take 0.1mL of the diluted solution and spread it on a plate containing a certain concentration of red tetrazolium medium, incubate at 25°C to observe the color change of the colonies, and pick out the red colonies in turn on the re-screening plate (containing a certain concentration of glycyrrhizic acid) Separation by streaking, streaking and purifying a single colony that can grow on a medium containing glycyrrhizinic acid, and finally shaking the flask for fermentation, detecting the content of GLA in the bacterial oil (as shown in Table 1), and screening to obtain the high-yield strain ME-L16.

[0035] The basic medium for primary...

Embodiment 2

[0042] Embodiment 2: Determination of optimal growth temperature and pH range of bacterial strains.

[0043] The ME-L16 strain is transferred to the PDA solid medium, cultured at 25-28° C. for 5-7 days, and the spores have matured. Pick 2 rings of spores with an inoculation loop and culture them in 50 mL of liquid seed medium at 25-28°C for 2 days. Insert 10% of the inoculum into the fermentation medium and cultivate for 6 to 8 days. Single factor experiments were carried out on the culture temperature and pH respectively, wherein the pH of the culture medium was adjusted to 4.5, 5.5, 6.5, 7.5, 8.5 respectively; 20, 23, 25, 28, 30°C. The dry weight was measured, and the content of GLA was detected by GC-MS at the same time. Analyze the optimal temperature and pH of the strains to ferment and produce GLA. It can be seen from Table 2 that the optimum pH of the bacteria is 5.5-7.5, and from Table 3 it can be seen that the optimum growth temperature of the bacteria is 25-28°C, ...

Embodiment 3

[0050] Embodiment 3: Laboratory culture of ME-L16 strain.

[0051] PDA medium: potato 200g / L, glucose 20g / L, agar 15g / L, add water to 1L, sterilize at 121°C for 20min. (Preparation method: peel the potatoes, cut into pieces and boil for 30 minutes, then filter with gauze, add sugar and agar, and make up water to 1000mL after melting.) After the PDA slant is cooled, insert the ME-L16 strain, and cultivate it at 25°C for 5 days. Mature spores are washed with sterile water, filtered through 6 to 8 layers of gauze, and placed in shake flasks for activation and culture. Shake flask activation medium contains the following components: glucose 30g / L, yeast extract 6g / L, KH 2 PO 4 3g / L, MgSO 4 ·7H 2 O 0.5g / L, the rest is water, pH 7.0, 50mL of activation medium is filled into a shaker flask with a liquid content of 250mL, activated at 120rpm, 25°C for 3 days. Then the activated bacterial strain was inserted into the fermentation medium, and the shake flask with a filling volume o...

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Abstract

The invention belongs to the technical field of biological engineering, and discloses Mortierella isabellina and application thereof to producing gamma-linolenic acid. The strain is named Mortierella isabellina ME-L16 through classification and is collected in China Center for Type Culture Collection with the collection number of CCTCC NO:M 2010195. The Mortierella isabellina is used for producing high-concentration gamma-linolenic acid by fermentation and is suitable for industrially producing the gamma-linolenic acid. Meanwhile, the invention discloses a method for screening strains for highly yielding the gamma-linolenic acid. Therefore, foundation is laid for further screening high-yield strains.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to Mortierella chrysogenum with high yield of gamma-linolenic acid and its application. Background technique [0002] γ-Linolenic acid (γ-Linolenic acid or Gamma-Linolenic acid, GLA for short) is all-cis 6,9,12-octadecatrienoic acid, molecular formula C 18 h 30 o 2 , is an omega-6 series polyunsaturated fatty acid, which is a colorless oily liquid and is easily oxidized in the air. It is an essential fatty acid in the human body, which is converted from linoleic acid in the human body, and then converted into double homolinolenic acid (Dihomo-γ-Linolenic acid, DGLA) and arachidonic acid (ARA), And then converted into prostaglandin E1 (PGE1), leukotrienes (LT) and prostacyclin I2 (PGI2). γ-linolenic acid has a wide range of physiological activities and obvious pharmacological effects. It can lower blood fat, resist thrombotic cardiovascular and cerebrovascular diseases, prev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 黄和纪晓俊黎志勇彭超丛蕾蕾任路静高振
Owner NANJING UNIV OF TECH
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