Bacterial strain for lowering water-soluble sugar in tobacco leaf as well as screening method, culture method and application

A water-soluble sugar and screening method technology, applied in the direction of microorganism-based methods, applications, biochemical equipment and methods, etc., can solve the problems of sugar-base ratio imbalance and poor quality of tobacco leaves, and achieve the reduction of acid gas in smoke and sensory quality Improve the effect of vigorous mycelium growth

Active Publication Date: 2017-02-22
CHINA TOBACCO SICHUAN IND CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a kind of abnormal Wickham yeast (Wickerhamomycesanomalus) that can reduce the water-soluble sugar in tobacco leaves to solve the problems of high content of water-soluble sugar in some tobacco leaves, serious imbalance of sugar-base ratio, and poor quality of tobacco leaves.

Method used

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  • Bacterial strain for lowering water-soluble sugar in tobacco leaf as well as screening method, culture method and application
  • Bacterial strain for lowering water-soluble sugar in tobacco leaf as well as screening method, culture method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Screening and isolation of abnormal Wickham yeast:

[0035] Take 10 g of soil samples (Jiangzhou Tobacco Field Soil, Liangshan, Sichuan) and put them in 90 mL enrichment medium, and culture them at 30°C for 96 hours. If any hyphae grow during the culture process, pick them out and discard them. Take 1 mL of the enriched culture solution, put it into a petri dish containing the separation medium that was melted and cooled to 50 °C, mix evenly, and after solidification, place it in a 30 °C constant temperature incubator for 48 hours. The grown colony was picked and inserted into the acclimatization medium, cultured at 30°C for 48 hours, and the culture was diluted and spread on the separation medium plate to obtain a single colony, which was isolated and purified by streaking to obtain Wickerhamomyces anomalus).

[0036] The enrichment medium is: 20.0g glucose, 10.0g peptone, 5g yeast extract, 5g lactic acid, pH 5.0, sterilized at 121°C for 20min,

[0037] Th...

Embodiment 2

[0044] Example 2: Preparation of seed solution of abnormal Wickham yeast:

[0045] The Y05 colony of Wickhamiana abnormalis obtained in Example 1 was expanded and produced to obtain Bacillus cereus (code Y15) seed solution.

[0046] Slant culture: inoculate the frozen-preserved strain of Wickerhamomyces anomalus Y05 on the bran juice slant medium, and culture at a constant temperature of 30° C. for 1-2 days. The bran juice slant medium is: add 150 mL of water to 25 g of bran, heat and boil for 30 minutes, filter, make up 100 mL of the filtrate, add 2 g of glucose, 0.5 g of yeast extract, and 1.5 g of agar.

[0047]Shake flask liquid culture: 1000mL of 1% glucose aqueous solution, sterilized by high pressure steam, inoculate the bacteria obtained from slant culture into the medium liquid, culture temperature: 30°C, shaker 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

[0049] The abnormal Wickham yeast seed solution obtained in Example 2 was added to tobacco leaves with a water content of 15% according to 30% of the weight of the tobacco leaves, and fermented at 30°C for 6 days. Compared with the unfermented blank tobacco leaves, the fermented tobacco leaves The total water-soluble sugar content in the medium was reduced by 60.5%, and the sensory evaluation results showed that the smoke coordination of the fermented tobacco leaves was better, the acidity was weakened, the sweetness returned, and the sense of satisfaction was enhanced.

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PUM

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Abstract

The invention discloses a bacterial strain for lowering water-soluble sugar in a tobacco leaf. The bacterial strain is wickerhamomyces anomalus; the bacterial strain is obtained through the following way of taking 9g to 10g of a soil sample, putting the soil sample in 90mL of an enrichment culture medium, afterwards, culturing the soil sample for 92h to 100h at 28 to 32 DEG C to obtain an enrichment culture solution, if a mycelium sends forth in a culture process, picking out and discarding the mycelium; taking 1 to 1.2mL of the enrichment culture solution, putting the enrichment culture solution into a culture dish loaded with an isolation culture medium which is cooled to be 48 to 52 DEG C after being fused, solidifying an obtained mixture, then putting the obtained mixture into a constant-temperature culture box at 28 to 32 DEG C, and culturing the obtained mixture for 45h to 50h; afterwards, selecting a growing bacterial colony, inoculating the bacterial colony into an acclimation culture medium, and culturing the bacterial colony in a standing manner for 45h to 50h at the temperature of 28 to 32 DEG C; next, diluting a culture in a gradient manner and coating the culture on an isolation culture medium slab, so as to obtain a single bacterial colony, and obtaining the bacterial strain through streaking and purification. By using the bacterial strain to treat the tobacco leaf which is high in sugar content and seriously disordered in sugar-nicotine ratio in a fermentation manner, 20 to 95 percent of water-soluble content can be lowered.

Description

technical field [0001] The invention relates to a microbial strain, in particular to an abnormal Wickham yeast (Wickerhamomyces anomalus) capable of reducing water-soluble sugar in tobacco leaves, a screening method, a culture method and the application of the strain in tobacco leaves. Background technique [0002] Tobacco leaves are the raw material for cigarette production, and the quality of tobacco leaves is directly related to the quality of cigarettes. The chemical conventional components of tobacco leaves are mainly: total plant alkaloids, water-soluble total sugars, reducing sugars, total nitrogen, chlorine and potassium, and also contain macromolecular compounds such as protein, starch, cellulose, pectin, etc. 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. The water-soluble sugar in tobacco leaves includes reducing sugar and non-reducing water-s...

Claims

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Application Information

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IPC IPC(8): C12N1/16A24B15/20C12R1/645
CPCA24B15/20C12N1/16C12N1/145C12R2001/645
Inventor 周容李东亮寇明钰
Owner CHINA TOBACCO SICHUAN IND CO LTD
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