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Process for extracting oxalate oxidase from bran of wheat

A technology of oxalate oxidase and wheat bran, applied in the direction of oxidoreductase, etc., can solve the problems of oxalate oxidase application limitation and unseen problems

Inactive Publication Date: 2005-03-23
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this way, the practical application of oxalate oxidase is limited
Many researchers try to reduce costs by immobilizing enzymes and enzyme electrodes, but there are no reports of their use in practice

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Take 1.5g of bran, first rinse with distilled water until the eluate is clear, then add 15mL of 50mmol / L Tris-HCl buffer solution with a pH of 7.5 to fully homogenate, centrifuge at 4000g for 20min at 4°C to collect the precipitate, and then Add the same buffer solution to rinse the precipitate until the eluate is clear, centrifuge at 4000g for 15 min to collect the precipitate, and rinse again with the same method. Suspend the precipitate obtained in the previous step with 15 mL of 50 mmol / L pH7.5 Tris-HCl buffer solution containing 1 mol / L NaCl, incubate in a water bath at 37 °C for 1 h, then centrifuge at 4000 g for 20 min at 4 °C, collect the precipitate, and use 15 mL Wash the precipitate with 50mmol / L Tris-HCl buffer solution of pH 7.5 without NaCl, collect the precipitate by centrifugation at 4000g for 15min, and wash again in the same way. Then suspend the precipitate with 15mL 50mmol / L Tris-HCl buffer solution of pH 7.5, incubate in a boiling water b...

Embodiment 2

[0013] Example 2: Take 1.5 g of bran, rinse it with tap water until the eluate is clear, then homogenize the bran with 15 mL of 50 mmol / L Tris-HCl buffer solution with a pH of 7.5, centrifuge at 4000 g for 20 min at 4°C, and collect the precipitate , then add 15mL of distilled water to rinse the precipitate until the eluate is clear, centrifuge at 4000g for 15 min to collect the precipitate, and use the same method to rinse again. Suspend the precipitate obtained in the previous step with 15 mL of Tris-HCl buffer solution containing 1 mol / L NaCl 50 mmol / L pH7.5, incubate in a 37°C water bath for 1 hour, then centrifuge at 4000g for 20 minutes at 4°C to collect the precipitate, add 15 mL of distilled water to rinse Wash the precipitate until the eluate is clear, centrifuge at 4000g for 15 minutes to collect the precipitate, and rinse again in the same way. Then suspend the above precipitate with 15mL 50mmol / L Tris-HCl buffer solution with pH7.5, keep it in a boiling water bath ...

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PUM

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Abstract

The invention is a method for extracting oxalate oxidase from wheat bran and an enzyme preparing technique, washing the wheat bran by water, then homogenizing by buffer solution, and centrifuging and collecting deposit; using the same buffer solution or distilled water to drop-wash the deposit until the eluent is clear and then collecting the deposit; successively suspending the deposit with buffer solution containing 1mol / L NaCl, preserving the heat during water bath for 1-1.5 h and then centrifuging and collecting deposit; using buffer solution or distilled water to drop-wash the deposit until the eluent is clear and then collecting deposit; adding butter solution in the deposit, making boiling water bath for 15-20 minutes and centrifuging and collecting deposit, using buffer solution or distilled water to drop-wash the deposit until the eluent is clear and then collecting deposit; then suspending the deposit by water solution containing pepsin, preserving the heat during water bath for 1-1.5 h and then centrifuging and collecting deposit, using buffer solution or distilled water to drop-wash the deposit until the eluent is clear; and finally using vacuum condense-drying system to eliminate the water in the deposit, thus obtaining the oxalate oxidase.

Description

technical field [0001] The invention relates to enzyme preparation technology, in particular to the discovery and preparation of oxalate oxidase in wheat bran. technical background [0002] Oxalate oxidase catalyzes the oxidation of oxalate to CO 2 and H 2 o 2 , was discovered by Zaleski and Reinhard while studying wheat flour, and later found that bacteria, mosses and many higher plants have oxalate oxidase activity. In practice, oxalate oxidase is a useful analytical enzyme that can be used to determine the content of oxalic acid in biological fluids or beverages, and it is simple, specific, fast and does not require special equipment, and can also exclude the interference of vitamin C . In addition, Malpass et al. (2002) also found that immobilized oxalate oxidase can not only degrade oxalic acid in mice, but also play an active role in inhibiting the formation of calcium oxalate crystals, which opens up a new way for the use of enzymes to trea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04
Inventor 刘娥娥彭新湘
Owner SOUTH CHINA AGRI UNIV
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