Method for extracting lipoxygenase from soybean whey waste water through inverse pH gradient

A technology of soybean whey wastewater and lipoxygenase, which is applied in the direction of oxidoreductase, can solve the problems of large-scale production and application, such as difficult breakthroughs and high costs, and achieve resource recycling, low cost, and high purity. high effect

Active Publication Date: 2019-06-07
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Although LOX has a wide range of uses in the fields of food, chemical industry, etc., it is directly extracted from soybeans and other raw materials, the cost is too high, and there is no industrially produced enzyme preparation; and the efficient expression of recombinant LOX by genetically engineered bacteria exists. Security and many other issues make it difficult to make major breakthroughs in large-scale production applications

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  • Method for extracting lipoxygenase from soybean whey waste water through inverse pH gradient
  • Method for extracting lipoxygenase from soybean whey waste water through inverse pH gradient
  • Method for extracting lipoxygenase from soybean whey waste water through inverse pH gradient

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Effect test

Embodiment 1

[0064] A method for extracting lipoxygenase from soybean whey wastewater against a pH gradient, comprising the steps of:

[0065] 1) Pretreatment of soybean whey wastewater

[0066] Take 1000mL soybean whey wastewater, centrifuge at 12000r / min for 10min, and keep the supernatant as soybean whey wastewater pretreatment liquid for use in experiments;

[0067] 2) Removal of impurity proteins in soybean whey wastewater pretreatment liquid

[0068] Using sodium hydroxide solution to adjust the pH of the soybean whey wastewater pretreatment solution to 8.0, and centrifuging to remove the precipitate;

[0069] 3) Extraction of lipoxygenase

[0070] Use sodium hydroxide solution to adjust the pH value of the supernatant obtained in step 2) to 6.0, remove the supernatant by centrifugation, dissolve the obtained precipitate in distilled water, and adjust the pH to neutral, which is the lipoxygenase crude enzyme solution, measure the enzyme activity and protein content in the crude en...

Embodiment 2

[0085] A method for extracting lipoxygenase from soybean whey wastewater against a pH gradient, comprising the steps of:

[0086] 1) Pretreatment of soybean whey wastewater

[0087] Take 1000mL of soybean whey wastewater, centrifuge at 12000r / min for 10min, and keep the supernatant as soybean whey wastewater pretreatment solution for use in experiments;

[0088] 2) Removal of impurity proteins in soybean whey wastewater pretreatment liquid

[0089] Using sodium hydroxide solution to adjust the pH of the soybean whey wastewater pretreatment solution to 8.0, and centrifuging to remove the precipitate;

[0090] 3) Extraction of lipoxygenase

[0091] Use sodium hydroxide solution to adjust the pH value of the supernatant obtained in step 2) to 6.0, remove the supernatant by centrifugation, dissolve the obtained precipitate in distilled water, and adjust the pH to neutral, which is the lipoxygenase crude enzyme liquid;

[0092] 4) The lipoxygenase crude enzyme solution is purif...

Embodiment 3

[0111] A method for extracting lipoxygenase from soybean whey wastewater against a pH gradient, comprising the steps of:

[0112] 1) Pretreatment of soybean whey wastewater

[0113] Take 1000mL of soybean whey wastewater, centrifuge at 12000r / min for 10min, and keep the supernatant as soybean whey wastewater pretreatment solution for use in experiments;

[0114] 2) Removal of impurity proteins in soybean whey wastewater pretreatment liquid

[0115] Using sodium hydroxide solution to adjust the pH of the soybean whey wastewater pretreatment solution to 8.0, and centrifuging to remove the precipitate;

[0116] 3) Extraction of lipoxygenase

[0117] Use sodium hydroxide solution to adjust the pH value of the supernatant obtained in step 2) to 6.0, remove the supernatant by centrifugation, dissolve the obtained precipitate in distilled water, and adjust the pH to neutral, which is the lipoxygenase crude enzyme liquid;

[0118] 4) Preliminary purification of lipoxygenase crude ...

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Abstract

The invention discloses a method for extracting lipoxygenase from soybean whey waste water through inverse pH gradient, and belongs to the technical field of utilization of waste water obtained duringfood processing. The method comprises the following steps of after the soybean whey waste water is pretreated, extracting the lipoxygenase by an inverse pH gradient method, firstly removing hybridprotein at the pH of isoelectric points of the lipoxygenase, then separating the lipoxygenase at the isoelectric points of the lipoxygenase, and then combining ultrafiltration with gel chromatography toobtain refined lipoxygenase. The soybean whey waste water is used for obtaining the lipoxygenase high in industrial value, and the extraction rate of the lipoxygenase is high. The lipoxygenase is highin purity and good in enzymatic activity. The method is convenient to operate, safe, effective, and low in cost, and can be used for development and utilization of the lipoxygenase in the soybean whey waste water in industrial use so as to realize source recycling. The method has notable economic benefits and social benefits.

Description

technical field [0001] The invention relates to the technical field of food processing wastewater utilization, in particular to a method for extracting lipoxygenase from soybean whey wastewater against a pH gradient. Background technique [0002] Soybean is an important crop with rich protein content. Soybean protein isolate processed from it is a refined soybean protein product with high protein purity and deep processing performance. It can be added to food production as an intermediate raw material and has a wide range of uses. [0003] Soybean protein isolate can be processed by alkali-soluble acid precipitation method, ultrafiltration concentration method, reverse micellar extraction method, reversed-phase high-performance liquid chromatography and other methods. Among them, the alkali-soluble acid precipitation method is relatively mature and widely used in the production of soybean protein isolate. Applied process method. However, when using the alkali-soluble acid p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02
Inventor 张志国
Owner QILU UNIV OF TECH
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