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Method for preparing nitrite reductase from flammulina velutipes

A technology of nitrite and reductase is applied in the field of food enzyme biology to achieve the effects of reducing content, high extraction efficiency and yield, and avoiding potential food safety hazards.

Pending Publication Date: 2017-03-01
NANJING UNIV OF FINANCE & ECONOMICS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation raw materials, efficient preparation methods and mechanism of action of nitrite reductase still need to be further explored.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0019] Example Preparation of nitrite reductase

[0020] Wash the by-products of Flammulina velutipes processing (mushroom roots, stems and defective mushrooms) thoroughly with running water or a washing machine, drain, cut into pieces, put them in a -20°C refrigerator for 12 hours, and then put them in a vacuum freeze dryer for 48 hours , after the freeze-dried sample was pulverized at low temperature (4° C.), the mushroom powder was passed through a 100-mesh sieve to obtain the mushroom powder. Dissolve the mushroom powder in distilled water according to the material-to-liquid ratio of 1:30, treat with 135W ultrasonic wave, adjust the pH to 5.0, add 400U / g mushroom powder weight of wall-lyzing enzyme, enzymatically hydrolyze for 2 hours at 35°C, adjust the pH to neutral, and centrifuge at 5000r / min After 30 minutes, collect the supernatant, add ammonium sulfate to 60% saturation, let it stand at 4°C for 2 hours, centrifuge at 5000r / min for 20 minutes, take the precipitate, d...

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PUM

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Abstract

The invention relates to a method for preparing nitrite reductase from flammulina velutipes and belongs to the technical field of a food enzyme biotechnology. The method comprises washing, draining and freeze-drying fresh flammulina velutipes, crushing the flammulina velutipes at a low temperature, screening the flammulina velutipes powder through a sieve of 60-100 meshes, carrying out extraction through an ultrasonic-assisted wall-dissolution enzyme method, carrying out salting-out, carrying out dialysis desalination, carrying out concentration to obtain a crude enzyme solution, carrying out separation purification through a DEAE-SepharoseFast Flow and dextrane gel G-150 chromatographic column, collecting a protein with enzyme activity, and carrying out dialysis, PEG20000 embedding and concentration, and freeze-drying to obtain nitrite reductase. The method provides a new idea for preparation of nitrite reductase, can be used in the field of fermented foods such as pickles, bacon and pickle development and reduces the content of nitrite in the product.

Description

[0001] 1. Technical field [0002] The invention relates to a method for preparing nitrite reductase from Flammulina velutipes, belonging to the field of food enzyme biotechnology. [0003] 2. Background technology [0004] Nitrite is one of the common chemical pathogenic sources. It is the precursor of strong carcinogen nitrosamines. After ingestion, it can react with secondary amines in the human body to form nitrosamines, which can induce various cancers; Excessive intake of nitrite can also cause methemoglobinemia, leading to asphyxia due to hypoxia. In recent years, due to the extensive use of nitrogen fertilizers and the use of industrial wastewater and domestic sewage to irrigate farmland, the nitrite pollution of agricultural land has intensified, resulting in excessive nitrite in agricultural products such as vegetables, fruits, grains, and feed; on the other hand, traditional pickled products, meat, etc. Products will accumulate or introduce nitrite during processing...

Claims

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Application Information

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IPC IPC(8): C12N9/06
CPCC12N9/0044C12Y107/01004
Inventor 杨文建王凡杜恒君胡秋辉裴斐马宁
Owner NANJING UNIV OF FINANCE & ECONOMICS
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