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Plant source amine oxidase as well as extraction method and application thereof

A technology of source amine oxidase and extraction method, applied in the field of active molecule extraction, can solve the problem of high biogenic amine content, and achieve the effects of simple preparation process, strong degradation ability and high safety

Pending Publication Date: 2022-05-27
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem of high biogenic amine content in food, to provide a plant-derived amine oxidase crude enzyme liquid and its preparation method, and to study its enzymatic properties

Method used

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  • Plant source amine oxidase as well as extraction method and application thereof
  • Plant source amine oxidase as well as extraction method and application thereof
  • Plant source amine oxidase as well as extraction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Extraction of amine oxidase from peanut

[0022] In this example, amine oxidase is extracted from peanuts, and the specific steps are as follows:

[0023] Select legume seeds with uniform size and plump grains, rinse them with distilled water, and soak them in distilled water for 1-3 days at 35°C. The soaked seeds were evenly sown in the germination bed at roughly the same interval, cultivated in a 35°C incubator, watered regularly, and kept the ambient humidity appropriate. The cultivation was terminated after the seeds had germinated 3-5 cm.

[0024] Remove the plant sprouts obtained in step (1), add 3 times of 1M potassium phosphate buffer (pH 7.0), and put them into a homogenizer to pulverize. Four layers of gauze were filtered, and the filtrate was centrifuged at 4°C and 9500 r / min for 15 min, and the supernatant was taken as the crude enzyme extract.

Embodiment 2

[0025] Embodiment 2: Determination of crude enzyme liquid properties of peanut amine oxidase extract

[0026] The assay method for amine oxidase activity is as follows:

[0027] Take the phosphate buffer solution of pH 7.0, add peroxidase solution, guaiacol solution, and mixed solution of nine biogenic amines.

[0028] Take the above mixed solution into a 96-well plate, set the reaction temperature of the microplate reader at 25-55°C, add the amine oxidase extract before putting on the machine, and measure the absorbance change of the reaction system at 470nm within 10-30min. Here, an amine oxidase activity unit (U) is defined as A per minute in the enzyme reaction system 470 Increase the amount of enzyme required to a value of 0.01.

[0029] The amine oxidase activity of the crude enzyme extract at 25, 30, 35, 40, 45, 50, 55, and 60 ℃ was compared under the condition of pH 7.0, and the curve was drawn with the highest value of 100%. The amine oxidase activity of the crude ...

Embodiment example 3

[0032] Implementation Case 3: Verifying the amine-reducing effect of crude amine oxidase extract in peanuts

[0033] The specific operation steps of the treatment group were as follows: take 2ml of crude enzyme solution and 1ml of biogenic amine mixture and mix it, the mixture was left for 24h at 40°C; centrifuged at 10000r / min for 5min; Chromatographic determination of biogenic amines.

[0034] The untreated group was treated for 24 hours with only the addition of buffer solution.

[0035] In this example, the crude amine oxidase enzyme solution extracted from peanut has obvious amine reducing effect. The results are shown in image 3 . The degradation rate of the crude enzyme extract from peanuts for phenethylamine reached 99.54%, and the degradation rate for the total amount of nine biogenic amines reached 61.83%

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Abstract

The invention discloses plant source amine oxidase as well as an extraction method and application thereof. The extraction method comprises the following steps: culturing plant tissues and extracting amine oxidase from the plant tissues. The enzyme provided by the invention is a plant-derived enzyme, so that the safety is higher; when common biogenic amine in food is taken as a substrate, obvious enzyme activity can be detected. And moreover, the amine-reducing enzyme has relatively strong degradation capability on several common biogenic amines in food. The method can be used in various foods with relatively high biogenic amine content; the high-histamine fish products, such as high-histamine fishes, shrimp paste, fish sauce, red wine, fermented vegetables, beer, ham, salted fish and spicy fermented tea, and common foods with high amino acid and protein contents, such as cereal fermented products, bean fermented products, milk fermented products, meat fermented products, fruit and vegetable fermented products, aquatic product fermented products and fermented tea, can be added into the high-histamine fish products, the cereal fermented products, the bean fermented products, the milk fermented products, the meat fermented products, the fruit and vegetable fermented products, the aquatic product fermented products and the fermented tea. And biogenic amines therein are degraded.

Description

technical field [0001] The invention relates to an extraction method of plant-derived amine oxidase and its application in a food system, which can be applied to the field of food processing and belongs to the technical field of active molecule extraction. Background technique [0002] Biogenic amines in food are usually produced by the degradation of free amino acids by amino acid decarboxylase in microorganisms, including histamine, tryptamine, putrescine, cadaverine, tyramine, phenethylamine, spermidine and spermine. In a certain period of time, when the intake of biogenic amines far exceeds the limit of biogenic amines that can be decomposed by the human body, it will cause changes in physiological functions, causing a series of adverse reactions such as headache, palpitations, rapid heartbeat, and facial flushing, which are serious and even endangering. life. Food safety problems caused by biogenic amines have received extensive attention. Biological control method re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/06
CPCC12N9/0022C12Y104/03006
Inventor 徐莹魏倩岚汪东风孙逊罗克马双
Owner OCEAN UNIV OF CHINA
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