Method for extracting myrosinase

An extraction method, sulfatase technology, applied in the field of agriculture, can solve the problem that the separation and purification method of myrosinase has not been studied and explained in detail, and achieve broad research value and market prospects, rich sources of raw materials, extraction method simple effect

Inactive Publication Date: 2011-03-30
CROPS BREEDING CULTIVATING INST SHANGHAI AGRI SCI ACAD
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Problems solved by technology

This technical solution is mainly used as a protection method for the activity of myrosinase, and

Method used

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  • Method for extracting myrosinase
  • Method for extracting myrosinase

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Embodiment approach 1

[0031] (1) By weight percentage, take 2 parts of frozen horseradish slices, add 4 parts of impurity-free deionized water (4 ℃ refrigerator precooling), the pH value of distilled water is adjusted between 6 ± 0.5, and then quickly ( within 2 minutes) into a homogenized pulp, put the whole pulp into the press, squeeze out all the juice with power hydraulic pressure, filter with 3 to 5 layers of medical gauze, and collect the filtrate; use qualitative analysis filter paper (medium speed) Carry out vacuum filtration on the filtrate, (vacuum degree -0.1~-0.01Mpa) and then collect the filtrate, enter the next processing step. The whole process requires to be carried out at low temperature (3-5°C).

[0032] (2) The collected filtrate was placed at 3-5° C. for 2 h, and the supernatant was subjected to cryogenic centrifugation (4° C., 6000 r / min, 15 min), and the precipitate was discarded. The supernatant was taken and refrigerated at 3-5°C until separation and purification.

[0033]...

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Abstract

The invention discloses a method for extracting myrosinase from horseradish, which comprises the following steps of: cleaning fresh horseradish, cutting into slices, precooling, adding clean distilled water with the pH of 5 to 7 and in an amount which is 2 times the weight of horseradish slices, homogenizing, filtering and collecting filtrate; standing, centrifuging, precipitating by using acetone, centrifuging again, suspending, cleaning and dehydrating by using the acetone, centrifuging and collecting the precipitate; and further freeze-drying the precipitate by using a low-temperature vacuum freeze-drying machine to obtain the myrosinase with high enzyme activity. The method is simple and more suitable for industrial production; and raw material sources are abundant.

Description

technical field [0001] The invention belongs to the technical field of agriculture and relates to a method for separating and purifying enzymes in plants. Background technique [0002] Myrosinase, commonly known as myrosinase (EC3.2.3.1), widely exists in nature. At present, the existence of myrosinase has been identified in more than 500 dicotyledonous plants belonging to 15 families, among which, the content of myrosinase is higher in cruciferous crops. [0003] At present, most of myrosinase are extracted from cruciferous vegetables, and the extraction method is not only cumbersome, but also has a relatively low yield. Jwanny et al proposed 2 kinds of myrosinases from radish root. They chopped 100g of fresh roots (dry weight: 6g), ground them with distilled water at 5°C, filtered the tissue homogenate with cheesecloth, then centrifuged at 4°C (13000r / min), and dialyzed the supernatant with water at 4°C for 2 days. Obtain the enzyme crude extract. For dialysate (NH4) ...

Claims

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Application Information

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IPC IPC(8): C12N9/24
Inventor 乔勇进王海宏方强陈召亮
Owner CROPS BREEDING CULTIVATING INST SHANGHAI AGRI SCI ACAD
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