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A halophilic archaea strain degrading nitrite and its application

A technology of halophilic archaea and nitrite, applied in the field of bioengineering, can solve the problems of not being screened, and achieve obvious effects and good salt tolerance

Active Publication Date: 2022-05-20
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using its denitrification effect, nitrite in high-salt food can be degraded into gaseous nitrogen compounds, so as to achieve the purpose of removing nitrite, but there is no good halophilic ancient strain for degrading nitrite. bacteria

Method used

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  • A halophilic archaea strain degrading nitrite and its application
  • A halophilic archaea strain degrading nitrite and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Screening of halophilic archaea

[0020] Take the self-made salted mustard pimples sold in the farmer's market in Anda City, Heilongjiang as the sample, weigh 10g of the sample, add 90mL of normal saline after crushing, shake fully for 1h and let it stand for 2h, take the supernatant and dilute it 10 times to 10 -1 ~10 -7 , draw 100 μL each and apply on the NHM solid medium. After culturing in a 37°C incubator for 7 days, select a single colony that is pink, round, and with neat edges, and streak three times continuously on the NHM solid medium to obtain pure colonies. strain JB33.

Embodiment 2

[0021] Example 2: Preliminary identification of denitrification function of halophilic archaea

[0022] Inoculate the screened bacterial strain JB33 into the screw cap tube (Durham tube) of the liquid denitrification medium with 1% inoculum after being activated in the NHM medium, culture it at 37°C, and observe whether the screw cap tube produces gas , to test whether the strain can metabolize nitrite to produce gas. It was observed that after 7 days of culture, air bubbles were produced in the screw cap tube, indicating that the strain JB33 could metabolize nitrite to produce gas.

Embodiment 3

[0023] Example 3: Morphological and molecular biological identification of halophilic archaea

[0024] The strain JB33 was inoculated on solid NHM medium and cultured at 37°C for 7 days. After a single colony was grown, the colony morphology and color were observed first, and then a single colony was picked for Gram staining: smear, fixation, crystal violet After primary staining, iodine solution mordant staining, ethanol decolorization, water washing and safranin counterstaining, drying and microscopic examination were carried out. Its biological characteristics are: negative Gram staining, colonies on NHM plates are pink, round, translucent, and neatly bordered.

[0025] Genomic DNA was extracted from the pure culture of strain JB33 according to conventional methods for molecular biology identification: the 16S rDNA sequence of the strain was used as a template for PCR amplification. The upstream primer of PCR amplification is 20F, its nucleotide sequence is shown in SEQIDN...

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Abstract

The invention belongs to the technical field of bioengineering, and in particular relates to a halophilic archaea strain for degrading nitrite and its application. The present invention found a Haloarcula A new species of the genus, named Haloarcula sp. JB33. The strain is a facultative aerobic denitrifying bacteria, which can grow at a salt concentration of 5-25% and a temperature of 20-52°C, and has good salt tolerance. The optimum growth temperature is 37°C. When the salt concentration is 18%, and the nitrite concentration is 100mg~500mg / kg, it still grows normally and degrades nitrite effectively, and the optimal degradation rate reaches 98.59% after 96 hours of degradation. Its degradation effect of nitrite is obvious. It can be used to degrade nitrite in food. It is especially used in the degradation of nitrite in high-salt pickled foods such as kimchi and ham.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a halophilic archaea strain for degrading nitrite and its application. Background technique [0002] Salted food is popular for its crisp and tender taste, salty and sweet, and is made by curing vegetables and salt. During the growth of vegetables, fertilization can lead to nitrogen accumulation and conversion into nitrate, which can be further converted into nitrite through microbial metabolism. The residual hazards of nitrite include: First, nitrite can oxyhemoglobin to form methemoglobin, releasing O 2 , leading to tissue hypoxia; the second is that nitrite can be converted into nitrosamines under certain conditions, and nitrosamines are a carcinogen, thereby inducing cancer. In the process of vegetable fermentation, lactic acid bacteria produce acid due to their metabolic process to reduce the pH of the environment. When the pH is lower than 4, nitrite ca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20A23L29/00C12R1/01
CPCA23L5/28A23L29/065C12R2001/01C12N1/205Y02A40/90
Inventor 魏巍胡信玉崔恒林朱琳
Owner JIANGSU UNIV
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