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Fermentation method for producing nitrite reductase

A technology of nitrite and reductase, which is applied in the direction of microorganism-based methods, methods using microorganisms, oxidoreductase, etc., can solve problems such as excessive nitrite, and achieve the effect of increasing production, broad prospects and safety advantages

Inactive Publication Date: 2019-04-26
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there has always been the problem of excessive nitrite in my country's traditional pickled vegetables and various meat enemas

Method used

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  • Fermentation method for producing nitrite reductase
  • Fermentation method for producing nitrite reductase
  • Fermentation method for producing nitrite reductase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Acclimatization and purification of strains.

[0035] 1. Acclimatization of strains

[0036] Add 0.01%~0.1% NaNO to the fermentation medium 2 , inoculated into the fermentation medium according to the inoculum amount of 2% of the volume of the fermentation broth, and cultured at 25-30° C. for 36-48 hours.

[0037] Fermentation medium: glucose 2g, peptone 1g, yeast powder 0.4g, dipotassium hydrogen phosphate 0.2g, beef powder 0.8g, magnesium sulfate 0.02g, sodium acetate 0.5g, triammonium citrate 0.2g, manganese sulfate 0.005g, spit Temperature-80 0.1g, distilled water 1L, pH6.2±0.2, sterilized by high-pressure steam at 121℃ for 30min.

[0038] 2. Purification of Strains

[0039] According to the conventional method for isolation of pure microorganisms, the domesticated strains are placed at 30°C for 48-72h. Inoculate into a new solid medium for selection of Lactobacillus plantarum, repeat at least 10 times, and purify colonies. Fermentation test was carried out in ...

Embodiment 2

[0043] Isolation and identification of purified strains:

[0044] (1) Bacteria and colony morphological characteristics of the purified strain

[0045] The morphological characteristics of the colony are: the colony is white, opaque, smooth, with neat edges and easy to pick. The biological characteristics of the strain were preliminarily identified by simple staining, Gram staining, capsule staining, and spore staining. Lambert-negative, non-capsulated, non-spore-forming, non-flagellate, such as figure 1 shown.

[0046] Genetic Traits:

[0047] The 16S rRNA whole gene base sequence of the purified strain is SEQ ID NO.1; the total 16S rRNA gene sequence of the strain is 1483bp (Genbank accession number: MH596000.1);

[0048] The strain has similarities with Lactobacillus plantarum, and its 16S rRNA sequence was systematically analyzed with Mega5.0 software, and the phylogenetic tree was constructed by Neigbor-Joining method to show that this strain and Lactobacillus plantarum...

Embodiment 3

[0050] (1) Culture medium preparation:

[0051] ① Solid seed medium: glucose 2g, peptone 1g, yeast powder 0.4g, dipotassium hydrogen phosphate 0.2g, beef powder 0.8g, magnesium sulfate 0.02g, sodium acetate 0.5g, triammonium citrate 0.2g, agar 2g, distilled water 1L, pH6.2±0.2, autoclaved at 121℃ for 30min;

[0052] ②Liquid seed medium: glucose 2g, peptone 1g, yeast powder 0.4g, dipotassium hydrogen phosphate 0.2g, beef powder 0.8g, magnesium sulfate 0.02g, sodium acetate 0.5g, triammonium citrate 0.2g, distilled water 1L, pH6 .2±0.2, 121°C high pressure steam sterilization for 30min;

[0053] ③Fermentation medium: glucose 2g, peptone 1g, yeast powder 0.4g, dipotassium hydrogen phosphate 0.2g, beef powder 0.8g, magnesium sulfate 0.02g, sodium acetate 0.5g, triammonium citrate 0.2g, manganese sulfate 0.005g, Tween-80 0.1g, distilled water 1L, pH6.2±0.2, sterilized by high pressure steam at 121℃ for 30min.

[0054] ④ Inoculate the strain on a solid seed medium according to a ...

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Abstract

The invention relates to the technical field of microbiology, enzyme engineering, fermentation engineering and biochemistry, in particular to a bacterial strain for producing nitrite reductase and a fermentation method thereof. After the bacterial strain of lactobacillus plantarum (the preservation number is CGMCC NO.1.12935) is domesticated and purified; the nitrite reductase is produced throughfermentation. After the bacterial strain is domesticated and purified, the yield of the nitrite reductase produced by bacteria through stable fermentation is improved; nitrite in food and feed can beeliminated. From the food safety angle, the lactobacillus nitrite reductase is developed for reducing and eliminating the residue of nitrite in food; wide prospects and safety advantages are realized.

Description

technical field [0001] The invention relates to the technical fields of microbiology, enzyme engineering, fermentation engineering and biochemistry, and relates to a bacterial strain for producing nitrite reductase and a fermentation method thereof. The nitrite reductase produced by the invention is mainly used in food industry, brewing, Fermentation, pharmaceutical and other industries. Background technique [0002] Nitrite is one of the main chemical pathogenic factors of food poisoning in my country in recent years. After nitrite is absorbed into the blood, it can make Fe in normal hemoglobin 2+ Oxidized to Fe 3+ , causing hemoglobin to lose its oxygen-carrying function, causing tissue hypoxia, and even hypoxia asphyxia. Nitrite is also a precursor of strong carcinogen nitrosamines, which is a potential carcinogenic hazard. It is closely related to the occurrence of gastric cancer, liver cancer and esophageal cancer. According to the epidemiological survey, the nitri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/06C12N1/36C12N1/02C12R1/25
CPCC12N1/02C12N1/36C12N9/0044C12Y107/01004
Inventor 张庆芳李美玉迟乃玉王晓辉于爽胡善松
Owner DALIAN UNIV
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