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Bacterial strain for reducing nicotine in tobacco leaves, screening method, culture method and application

A technology of tobacco leaves and bacterial strains, applied in the field of microbial strains, can solve the problems of poor quality and usability of tobacco leaves, imbalance of sugar-base ratio, etc., and achieve the effects of reducing pungent sensation, improving sensory quality and coordination

Active Publication Date: 2019-08-09
CHINA TOBACCO SICHUAN IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problems of high nicotine content in some stored tobacco leaves, serious imbalance of sugar-alkaline ratio, and poor quality and usability of tobacco leaves, the present invention provides a Bacillus cereus that can reduce nicotine in tobacco leaves;

Method used

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  • Bacterial strain for reducing nicotine in tobacco leaves, screening method, culture method and application
  • Bacterial strain for reducing nicotine in tobacco leaves, screening method, culture method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Isolation and identification of Bacillus cereus

[0034] Take 10g of soil sample (Liangshan Jiangzhou tobacco field soil in Sichuan) and put it in 90mL enriched medium, and cultivate it at 30℃ for 48h. Take 1 mL of the enriched culture solution, put it into a petri dish containing a separation medium that is melted and cooled to 50°C, mix well, and place it in a 30°C constant temperature incubator for 7 days after solidification. Pick the growing colonies into the domestication medium, culture at 30°C and 120rpm for 96 hours, spread the culture on the separation medium plate with gradient dilution to obtain a single colony, and obtain the Bacillus cereus strain (Bacillus cereus) through streaking separation and purification. cereus).

[0035] The enrichment medium is: peptone, 5.0g; NaCl, 1.0g; K 2 HPO 4 , 1.0g; agar, 1.5g; the pH of the enrichment medium is 7.0, sterilize at 121°C for 30 min, add 2.0 g of nicotine after cooling.

[0036] The separation medium is:...

Embodiment 2

[0042] Example 2: Preparation of Bacillus cereus seed liquid

[0043] The Bacillus cereus colony obtained in Example 1 was expanded and produced to obtain the Bacillus cereus seed liquid.

[0044] A. Slant culture: Peptone 10g, beef extract 3g, sodium chloride 5g, agar 15g, distilled water 1000mL, pH value 7.0~7.2, inoculate the frozen Bacillus cereus strain (Bacillus cereus) on the slant medium, 30 Incubate at a constant temperature for 1-2 days.

[0045] B. Shake flask liquid culture: peptone 10g, beef extract 3g, sodium chloride 5g, agar 15g, distilled water 1000mL, pH value 7.0-7.2, autoclave sterilization, inoculate the bacteria in step A into the medium liquid, Cultivation temperature: 30℃, shaker rotation speed 140-160r / min, shake flask culture 18-24h, cultivate until the number of bacteria reaches 10 10 Above, the seed liquid is obtained.

Embodiment 3

[0047] The Bacillus cereus seed liquid obtained in Example 2 was added to tobacco leaves with 15% moisture content according to 30% of the weight of the tobacco leaves, and fermented for 6 days at 25°C. Compared with the unfermented blank tobacco leaves, the fermented tobacco leaves were The alkali content was reduced by 24.9%, and the sensory evaluation results showed that the strength of fermented tobacco leaves was significantly reduced, the irritation was significantly reduced, and the smoke was more mellow.

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Abstract

The invention discloses a bacterial strain for reducing nicotine in tobacco leaves. The bacterial strain is Bacillus cereus, which is obtained by taking 9-10 g of soil samples in 90 mL of enrichment medium, and then placing them at a temperature of 28-32°C Cultivate for 40-50 hours to obtain the enriched culture solution; take 1-1.2 mL of the enriched culture solution, put it into a petri dish containing the separation medium that has been melted and cooled to 48-52°C, and place it in a refrigerator at a temperature of 28-32°C after solidification. Cultivate in a constant temperature incubator for 6 to 8 days; then select the grown colony and insert it into the acclimatization medium, and cultivate it for 90 to 110 hours at a temperature of 28 to 32 ° C and a rotation speed of 100 to 150 rpm; then apply gradient dilution of the culture on The media plates were dissociated to obtain single colonies, which were purified by streaking. Using the bacterial strain to ferment and treat tobacco leaves with high nicotine content and seriously imbalanced sugar-alkali ratio can reduce the nicotine content by 20-40%.

Description

Technical field [0001] The present invention relates to a microbial strain, in particular to a Bacillus cereus (Bacillus cereus) capable of reducing nicotine in tobacco leaves, a screening method, a culture method and application of the strain in tobacco leaves. Background technique [0002] Tobacco leaf is the raw material for cigarette production, and the quality of tobacco leaf is directly related to the quality of cigarettes. The conventional chemical components of tobacco leaves are mainly: total plant alkaloids, total water-soluble sugars, reducing sugars, total nitrogen, chlorine and potassium. It also contains protein, starch, cellulose, pectin and other macromolecular compounds. The content of these components in tobacco leaves And the proportional relationship between certain components has an important impact on the quality of tobacco leaves. Nicotine, also known as Nicotine, is the main component in tobacco alkaloids. The content of nicotine in flue-cured tobacco is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A24B15/20C12R1/085
CPCA24B15/20C12N1/20C12N1/205C12R2001/085
Inventor 周容李东亮寇明钰
Owner CHINA TOBACCO SICHUAN IND CO LTD
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