Aspergillus parasiticus and applications thereof in preparation of nitrite reductase, nitrite reductase gene and genetically engineered bacterium

A technology of parasitic Aspergillus and nitrite, applied in the field of microorganisms, can solve the problems of unsuitable breeding water and food treatment, low efficiency, secondary pollution, etc., and achieve fast and reliable degradation, good degradation effect, high efficiency and low energy consumption.

Inactive Publication Date: 2015-01-28
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many problems in these methods. Among them, reverse osmosis, electrodialysis, and ion exchange all concentrate nitrite in the medium or waste liquid, which is easy to cause secondary pollution, and has low efficiency and large investment. It is suitable for the treatment of industrial water; although the chemical reduction method can reduce nitrite to harmless nitrogen, the reaction needs to be carried out at a higher temperature, and the efficiency is low, so it is not suitable for the treatment of aquaculture water and food; Although the chemical oxidation method has the advantages of simple equipment and relatively low treatment costs, it is only suitable for the treatment of deep underground water with low nitrite content and less treatment capacity, and is not suitable for the treatment of aquaculture water and food

Method used

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  • Aspergillus parasiticus and applications thereof in preparation of nitrite reductase, nitrite reductase gene and genetically engineered bacterium
  • Aspergillus parasiticus and applications thereof in preparation of nitrite reductase, nitrite reductase gene and genetically engineered bacterium
  • Aspergillus parasiticus and applications thereof in preparation of nitrite reductase, nitrite reductase gene and genetically engineered bacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Screening, cultivation, identification and performance testing of Aspergillus parasitic strain JFS

[0032] 1. Screening of Aspergillus parasitic strain JFS:

[0033] (1) Material

[0034] a. Samples: 20 samples of water, soil, dead wood, rabbit dung and sheep dung collected from Chengdu, Sichuan, Yibin, and Chongqing Rongchang.

[0035] Sample processing and storage: The collected soil samples, dead wood, rabbit dung and sheep dung were all dried in a cool and dry place immediately after being collected, and stored in a clean kraft paper envelope after drying. The water sample is used directly after collection (fresh water is used).

[0036] b. Medium:

[0037] Nitrite selection medium (unit g / L):

[0038]

[0039] Cha's medium (unit g / L):

[0040] All the above-mentioned media are prepared in 1×10 5 Sterilize for 25 min under Pa.

[0041] c. Reagents and instruments:

[0042] Reagents: potassium ferrocyanide, zinc acetate, p-aminobenzenesulfonic acid, naphthalene ethyl...

Embodiment 2

[0119] Example 2. Application of Aspergillus parasiticus in the preparation of nitrite reductase gene

[0120] The preparation steps of nitrite reductase gene are as follows:

[0121] (a) Transfer the strain JFS from Cha's medium slant to Cha's plate or culture on Cha's slant at 30°C for 3-7 days, or transfer to Cha's seed medium at 30°C, 150r / min After culturing for 3 to 8 days, centrifuge or filter to collect bacteria and sporophytes.

[0122] (b) DNA extraction of the collected bacteria and sporophytes: the genomic DNA of strain JFS is extracted.

[0123] (c) Design degenerate primers based on the conserved regions of the amino acid sequence of nitrite reductase of various organisms in the Genbank protein database, and use the genomic DNA of strain JFS as a template to amplify the DNA fragment of nitrite reductase in strain JFS, Then sequence. The specific gene preparation can use the existing technology, which will not be repeated here.

[0124] In the preparation of the nitrite ...

Embodiment 3

[0125] Example 3 Application of Aspergillus parasiticus in the preparation of nitrite reductase

[0126] The strain JFS was transferred from the Cha's medium slant to Cha's seed medium. After culturing at 30°C and 150r / min for 2 days, it was transferred to the fermentation medium at 30°C, 150r / min with an inoculum of 3-10%. Cultivate for 3-9 days, centrifuge or filter, collect bacteria and sporophytes, rupture the cell wall of strain JFS with ultrasonic or liquid nitrogen grinding or glass bead grinding or use the above three methods at the same time, and then extract, Salting out, chromatography and other methods to purify nitrite reductase. The specific preparation method of nitrite reductase can adopt the existing technology, which will not be repeated here.

[0127] The Aspergillus parasitica of the present invention is used in the preparation of nitrite reductase. Because the Aspergillus parasitica of the present invention is easy to cultivate and has low cultivation cost, th...

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Abstract

Aspergillus parasiticus and applications thereof in preparation of nitrite reductase, a nitrite reductase gene and a genetically engineered bacterium containing the nitrite reductase are disclosed. The accession number of the aspergillus parasiticus is CCTCC M2013662. The aspergillus parasiticus has characteristics of high efficiency, low energy consumption, good nitrite degradation effects, and rapid and reliable degradation is provided, and can be well applied in preparation of the nitrite reductase, preparation of the nitrite reductase gene and preparation of the genetically engineered bacterium containing the nitrite reductase.

Description

Technical field [0001] The invention belongs to the technical field of microorganisms, and relates to an Aspergillus and its application, in particular to an Aspergillus parasitic, and the preparation of nitrite reductase, nitrite reductase gene preparation and genes containing nitrite reductase Application in the preparation of engineering bacteria. Background technique [0002] Nitrite is one of the most common nitrogen compounds in nature, and its sources are also very wide. Nitrite is a harmful chemical substance. Nitrite interacts with human blood to form methemoglobin, which makes the blood lose oxygen carrying Function, make people hypoxic poisoning, light dizziness, heart palpitations, vomiting, lip bruising, severe, unconsciousness, convulsions, shortness of breath, untimely rescue can endanger life. Not only that, nitrite can also interact with secondary amines in the human body to form nitrosamines, which are carcinogenic, teratogenic, and mutagenic substances in the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/06C12N15/53C12N15/70C12N15/81C12R1/66
CPCC12N9/0044C12Y107/02002C12N1/145C12R2001/66
Inventor 刘冰花杨林蒲小龙罗亚雄袁管明
Owner CHENGDU UNIV
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