Bacillus altitudinis J54 and application thereof

A technology of Bacillus highlandi and Geobacillus, applied in the field of microorganisms, can solve the problems of few methods for TSNAs degradation, and achieve the effects of good stress resistance, good colonization effect and low cost

Active Publication Date: 2020-01-21
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are relatively few methods for the degradation of TSNAs

Method used

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  • Bacillus altitudinis J54 and application thereof
  • Bacillus altitudinis J54 and application thereof
  • Bacillus altitudinis J54 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] - Bacillus highlander ( Bacillus altitudinis ) Acquisition and Identification of J54

[0075] (1) Bacillus highlander ( Bacillus altitudinis ) of J54 to obtain

[0076] A. Separation: Leaf samples of flue-cured tobacco K326 were collected from Yuxi City, Yunnan Province. Wash 5 g of the collected tobacco leaves with sterile water, sterilize the surface, cut them into pieces and place them in potassium phosphate buffer, treat them with ultrasonic waves for 30 min, and filter to remove the tobacco leaves. membrane, and then ultrasonically eluted the bacterial cells. The collected bacteria were evenly spread on the NIM medium with nicotine as the only carbon source, and single colonies were picked and isolated.

[0077] The culture condition is 30°C, and the time is 48-72h;

[0078] The composition of the separation medium is calculated in g / L: Na 2 HPO 4 6 g / L, KH 2 PO 4 3 g / L, NH 4 Cl 1 g / L, NaCl 0.5 g / L, MgSO 4 0.12 g / L, CaCl 2 0.1 g / L, nicotine 0.5 g / L,...

Embodiment 2

[0096] - Bacillus highlander ( Bacillus altitudinis ) Degradation experiment of J54 on TSNAs during tobacco leaf curing

[0097] experimental method:

[0098] a. Fermentation: First, Bacillus alba ( Bacillus altitudinis ) J54 was inoculated on the slant medium to obtain fermented seeds;

[0099] The culture conditions are: 30°C, time 48h;

[0100] The components of the slant medium are calculated in g / L: tryptone 10g / L, yeast extract 5 g / L, NaCl 5 g / L, agar 15.0g / L, pH7.4;

[0101] Then the fermented seeds are inoculated into the seed culture solution, the inoculation amount is 1v / v%, and the liquid volume of the shake bottle is 12v / v%, so as to obtain the fermentation agent;

[0102] The culture conditions were: 30°C, 48 h;

[0103] The components of the seed culture solution are calculated in g / L: tryptone 10g / L, yeast extract 5 g / L, NaCl 5 g / L, pH7.4;

[0104] The effective number of viable bacteria of the obtained fermentation agent is 1.1×10 9 individual / mL.

[01...

Embodiment 3

[0112] - Bacillus highlander ( Bacillus altitudinis ) Degradation experiment of J54 on TSNAs in cured tobacco leaves

[0113] experimental method:

[0114] A, fermentation: method is with embodiment 2

[0115] The effective number of viable bacteria of the obtained fermented bacterial agent is 1.2×10 9 pieces / ml.

[0116] b. Inoculation: During the tobacco leaf preparation process, the whole plant was cut and harvested, and the tobacco leaf preparation experiment was carried out by the half-leaf method. Half of the leaves were sprayed with fungicide, and the other half were sprayed with sterile water as the control (CK). Before the preparation, the fermentation agent is sprayed according to 4% of the weight of the tobacco leaves.

[0117] c. Degradation: Put the tobacco leaves sprayed with fermented bacterial agent and the control tobacco leaves in an incubator, at 30°C and a relative humidity of 80%, after air curing for 45 days, the middle leaves were sampled to measure...

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Abstract

The invention discloses Bacillus altitudinis J54 and application thereof. The Bacillus altitudinis is Bacillus altitudinis J54, is preserved in the China Center for Type Culture Collection, and has apreservation number of CCTCC No. M2019667. Separation is performed on tobacco leaves, and then screening and purification are performed so as to obtain the Bacillus altitudinis J54; through processesof fermentation, inoculation and degradation, nitrosamine (TSNAs) specific in tobaccos can be degraded effectively in the process of tobacco leaf processing or pipe tobacco preparation, and the degradation rate can reach 5.2-46.6%, so that the quality of the tobacco leaves is improved; and the bacterial strain has good resistance to stress, easy acquisition, convenient use and broad market application prospects.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a highland bacillus J54 and an application thereof. Background technique [0002] Tobacco-specific nitrosamines (TSNAs) are one of the most important harmful substances in tobacco and tobacco products. It is a class of carcinogens produced during the drying, processing, fermentation and burning of tobacco. There are serious effects. TSNAs mainly include 4-(N-nitrosomethylnitrogen)-1-(3-pyridyl)-1-butanone (NNK), N-nitrosonornicotine (NNN), N-nitroso Anatabine (NAT) and N-nitrosobasine (NAB) are the four most abundant TSNAs identified. Among them, NNK and NNN are the most carcinogenic. The formation of TSNAs in tobacco can be regulated by changing tobacco type, tobacco tissue, cultivation method, preparation method, microbial community, nitrogen fertilizer application rate, nitrate reductase and nitrosase activity and other factors. However, there are relati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A24B3/12A24B15/18C12R1/07
CPCC12N1/20C12N1/02A24B3/12A24B15/18C12R2001/07C12N1/205
Inventor 马俊红卢灿华盖晓彤雷丽萍莫笑晗余清姜宁夏振远
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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