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
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[0025] Implementation Example 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....
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[0030] Implementation example 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.
Example Embodiment
[0036] Implementation Example 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 eva...
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