Method for extracting pyrified isomerase of linoleic acid from strain of producing isomerase of linoleic acid

A technology for linoleic acid isomerase and purification of linoleic acid, applied in the directions of isomerase, fermentation, etc., can solve the problems of toxic solvents, complex isomer composition, complicated extraction and purification process, etc. The effect of reducing enzyme activity loss and high cell fragmentation rate

Active Publication Date: 2007-12-26
中农科合肥食品营养与健康创新研究院有限公司
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AI Technical Summary

Problems solved by technology

Although the process is simple and the requirements for equipment are low, the product contains a large amount of toxic solvents, the composition of isomers is complex, and the downstream extraction and purification process is complex, so there are certain limitations in the application in the food and pharmaceutical industries

Method used

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  • Method for extracting pyrified isomerase of linoleic acid from strain of producing isomerase of linoleic acid
  • Method for extracting pyrified isomerase of linoleic acid from strain of producing isomerase of linoleic acid
  • Method for extracting pyrified isomerase of linoleic acid from strain of producing isomerase of linoleic acid

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Embodiment 1

[0039] Embodiment 1, extraction and purification of linoleic acid isomerase

[0040] 1. Obtaining of linoleic acid isomerase fermentation broth

[0041] The fermentation medium used in this embodiment is prepared according to the following method:

[0042] (1) Preparation of sunflower seed oil emulsion: 300mg of sunflower seed oil (Shanghai Jiage Food Co., Ltd., Duoli brand 100% pure sunflower seed oil), 0.36ml of Tween-80 (Tween-80), 5ml of deionized water , ultrasonic emulsification (25° C., ultrasonic frequency 40 KHz) for 10 min, and filter sterilization with a 0.22 μm microporous membrane to obtain a sunflower oil emulsion.

[0043] (2) Preparation of skim milk medium: 12 g of skim milk powder (Sanyuan Dairy Co., Ltd.) was dissolved in 100 ml of water, and sterilized at 5° C. for 10 minutes for later use.

[0044](3) Preparation of fermentation medium: the sunflower oil emulsion of (1) was added to the skim milk medium of (2), so that the final concentration of sunflowe...

Embodiment 2

[0058] Embodiment 2, extraction and purification of linoleic acid isomerase

[0059] 1. Obtaining of linoleic acid isomerase fermentation broth

[0060] The fermentation medium used in this embodiment is prepared according to the following method:

[0061] The fermentation medium used in this embodiment is prepared according to the following method:

[0062] (1) Preparation of sunflower seed oil emulsion: 400 mg of sunflower oil (Shanghai Jiage Food Co., Ltd., Duoli brand 100% pure sunflower oil), Tween-80 (Tweenum 80) 0.44 ml, deionized water 5 ml, Ultrasonic emulsification (25° C., ultrasonic frequency 40 KHz) for 10 minutes, and filter sterilization with a 0.22 μm microporous membrane to obtain sunflower oil emulsion.

[0063] (2) Preparation of skim milk medium: 12 g of skim milk powder (provided by Beijing Sanyuan Dairy Co., Ltd.) was dissolved in 100 ml of water, and sterilized at 5° C. for 10 minutes for later use.

[0064] (3) Preparation of fermentation medium: the...

Embodiment 3

[0078] Embodiment 3, extract and purify linoleic acid isomerase

[0079] 1. Obtaining of linoleic acid isomerase fermentation broth

[0080] The fermentation medium used in this embodiment is prepared according to the following method:

[0081](1) Preparation of sunflower seed oil emulsion: 600mg of sunflower seed oil (Shanghai Jiage Food Co., Ltd., Duoli brand 100% pure sunflower seed oil), 0.5ml of Tween-80 (Tween-80), 5ml of deionized water , ultrasonic emulsification (25° C., ultrasonic frequency 40 KHz) for 10 min, and filter sterilization with a 0.22 μm microporous membrane to obtain a sunflower oil emulsion.

[0082] (2) Preparation of skim milk medium: 12 g of skim milk powder (Sanyuan Dairy Co., Ltd.) was dissolved in 100 ml of water, and sterilized at 115° C. for 10 minutes for later use.

[0083] (3) Preparation of fermentation medium: the sunflower oil emulsion of (1) was added to the skim milk medium of (2), so that the final concentration of sunflower oil was 6...

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Abstract

This invention discloses a method for extracting linoleic acid isomerase from bacteria generating linoleic acid isomerase. The method comprises: (1) pulverizing and homogenizing the bacteria generating linoleic acid isomerase, centrifuging and collecting the supernatant containing crude enzyme liquid; (2) fractionally precipitating with (NH4)2SO4 with a saturation degree of 40-90%, collecting the precipitate, and suspending with K3PO4 buffer to obtain crude enzyme liquid; (3) loading crude enzyme liquid on an ion exchange column, eluting with 0.1-0.7 mol / L NaCl solution, and collecting the eluate with linoleic acid isomerase activity; (4) dialyzing with 0.08-0.15 mol / L K3PO4 buffer (pH = 5.5-7.0) to remove salts; (5) loading on a gel filtration column, and eluting with 0.08-0.15 mol / L K3PO4 buffer (pH = 5.5-7.0) to obtain linoleic acid isomerase.

Description

technical field [0001] The invention relates to a method for extracting and purifying linoleic acid isomerase from linoleic acid isomerase producing bacteria. Background technique [0002] Conjugated linoleic acid (Conjugated Linoleic Acid, CLA) is a very interesting nutritional supplement. A large number of in vitro and in vivo experiments have proved that CLA can induce energy utilization and cause weight loss without storing fat; it can alleviate such as anorexia , proteolysis, and other side effects of immune reactions; it has anti-mutation, inhibits the synthesis of proteins and nucleic acids in cancer cells, and in addition, CLA can also improve certain physiological states of humans, such as increasing the level of vitamin A in the body. Since CLA has rich nutritional value and its content in food is very low, how to prepare CLA with low cost and high purity has become the focus of research. The commercial production method of CLA is the alkali isomerization method of...

Claims

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Application Information

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IPC IPC(8): C12N9/90C12P7/64
Inventor 吕加平王丽敏任星环江均平
Owner 中农科合肥食品营养与健康创新研究院有限公司
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