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Debaryomyces hansenii strain and application thereof

A technology of yeast strains and seeds, applied in the field of microorganisms, can solve the problems of not meeting market demand, human health hazards, limited production, etc., and achieve significant industrial application value, low cost, and the effect of reducing miscellaneous gas and irritation

Active Publication Date: 2021-09-07
CHINA TOBACCO SICHUAN IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Synthetic fragrances are unnatural and can cause harm to human health
Natural spices are mainly derived from animals, plants and microorganisms, among which animal and plant spices have limited resources and limited output, which cannot meet the growing market demand
At present, there is no report on the screening of aroma-producing Debaria hansenii from tobacco leaves and its use in tobacco fermentation for flavor enhancement and production of natural tobacco flavors

Method used

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  • Debaryomyces hansenii strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Enrichment, purification and screening of strains:

[0027] Take 5 g of flue-cured tobacco leaves (Sichuan Panzhihua tobacco leaves) in 100 mL enrichment medium, place them in a shaker at 28° C., and culture at 150 rpm for 36 hours. Take 1 mL of the culture solution and place it in a petri dish, mix it evenly with the separation medium cooled to 50°C, and culture it at 28°C for 48 hours after solidification. The growing colonies were picked, inserted into the isolation medium, and cultured at 28° C. and 150 rpm for 36 hours. The culture was diluted in a gradient and spread on the separation medium plate to obtain a single colony, which was inserted into the separation medium, and the aroma-producing culture was screened by smelling the aroma. The culture with aroma was serially diluted and spread on the separation medium plate to obtain a single colony, which was separated by streaking to obtain Debaryomyces hansenii, numbered SCT-Y1.

[0028] The enrichment medium is...

Embodiment 2

[0031] ITS molecular identification and biological characteristics of the strain:

[0032] Use primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') to carry out PCR amplification of the ITS region molecular fragment of the strain, and then perform sequencing after agarose gel electrophoresis detection, and obtain the ITS sequences of strains.

[0033] Described Debaryomyces hansenii culture form is described as follows:

[0034] The surface of the colony on the potato dextrose agar medium is relatively wet, the shape is large and raised, creamy in color, with smooth edges, and the color of the front and back of the colony is the same.

[0035] Comparing the sequence with the ITS gene sequences of some strains registered in GenBank, the homology reached 99%. Combined with the morphological identification results, it was shown that the strain SCT-Y1 belonged to Debaryomyceshansenii.

Embodiment 3

[0037] Debaria hansenii was inoculated in potato dextrose medium, placed in a constant temperature shaker at 28°C, 200rpm, and cultured for 36h. Take the inoculum amount of 2% by volume of the seed solution, inoculate it in potato glucose medium, and place it in a constant temperature shaker at 28° C. and 150 rpm for 36 hours. The culture solution: water: tobacco leaves are evenly sprayed on the surface of the flue-cured tobacco leaves according to the mass ratio of 1:1:8, placed in a constant temperature and humidity box at 35°C and 75% relative humidity for 1 day, and the treated tobacco leaves are placed at 60°C Drying under conditions to a moisture content of 12% to obtain fermented tobacco leaves.

[0038] Potato glucose medium: 200.0g potato; 20.0g glucose; 1000mL tap water; natural pH; sterilized at 115°C for 20min.

[0039] The fermented tobacco leaves are cut into shreds and then rolled into cigarette sticks. The sensory quality evaluation is carried out by professio...

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Abstract

The invention discloses a debaryomyces hansenii strain and application thereof. The aroma-producing debaryomyces hansenii strain screened from tobacco leaves or fermentation liquor of the aroma-producing debaryomyces hansenii strain is inoculated to the surfaces of the tobacco leaves, and fermentation is performed under certain conditions, so that the quality of the tobacco leaves can be remarkably improved, and the fermented tobacco leaves have the characteristics that the sweet feeling is remarkably improved, the smoke is soft, and offensive odor and irritation are reduced, and the like.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of Debaria hansenii and its application. Background technique [0002] The lack of flavor of tobacco can be made up by adding tobacco flavors and fragrances. Tobacco flavors are mainly divided into two categories: synthetic and natural flavors. Synthetic fragrances are unnatural and can be harmful to human health. Natural spices are mainly derived from animals, plants and microorganisms, among which animal and plant spices have limited resources and limited output, which cannot meet the growing market demand. The preparation of natural spices by microbial fermentation has the characteristics of short production cycle, not affected by climate, mild reaction conditions, high selectivity and high efficiency, and has attracted more and more attention. [0003] Screening specific aroma-producing microorganisms from the surface of alcoholized tobacco leaves to produc...

Claims

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Application Information

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IPC IPC(8): C12N1/16A24B3/12A24B3/14A24B15/20C12R1/645
CPCC12N1/16A24B3/12A24B3/14A24B15/20Y02E50/10
Inventor 张倩颖李东亮罗诚蔡文耿宗泽马亚萍
Owner CHINA TOBACCO SICHUAN IND CO LTD
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