Debaryomyces hansenii with good fermentation performance and aroma production function and screening method thereof
A technology for yeast and fermented sausage, applied in the field of microorganisms, can solve the problem of aroma-producing yeast being in its infancy and the like
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Embodiment 1
[0026] Example 1: Collection of traditional air-dried sausage samples and separation and purification of yeast
[0027] Traditional air-dried sausage samples were collected from 5 regions in Northeast China (Harbin, Suihua, Daqing, Hegang, and Mudanjiang), and 10 g of minced air-dried sausage samples were added to 90 mL of 0.85% sterile saline (w / v). Shake on a shaker for 30 minutes, after 10-fold serial dilution, take 100 μL in the ultra-clean workbench and spread it on a YPD agar plate supplemented with 100 mg / L chloramphenicol, and place it in a microbial incubator at 28°C for 48 hours. Select a YPD agar plate with 30-300 colonies on the plate, pick independent colonies with different shapes on the plate that conform to the morphological characteristics of yeast colonies, and further purify them by streaking on a new YPD agar plate until a pure culture is obtained. , the obtained strains were stored in 20% glycerol at -80°C for future use.
Embodiment 2
[0028] Embodiment 2: the identification of yeast
[0029] The screened strains were subjected to morphological analysis and ITS rRNA sequence identification. The amplification primers used were ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). The results were compared on NCBI to identify As a result, 46 strains of yeast were found to be isolated.
[0030] Such as figure 1 Shown, the morphological characteristics of described bacterial strain SH4 are: the bacterium colony that grows on YDP agar plate is milky white, bacterium colony spherical protrusion, smooth and glossy surface, neat edge, opaque, easy to provoke (see figure 1 ).
[0031] The ITS rDNA sequencing result of the strain SH4 provided by the present invention is compared with the existing sequence in NCBI data, and it is confirmed that the strain SH4 is Debaryomyces hansenii.
Embodiment 3
[0032]Example 3: Screening of yeast with excellent fermentation properties
[0033] (1) Screening of acid-resistant, salt-resistant, and nitrite-resistant yeast: 46 yeast strains were divided into 10 6 CFU / mL were respectively inoculated into YPD liquid medium with different pH gradients (4.0 and 5.0), different sodium chloride concentrations (4% and 6%) and different sodium nitrite concentrations (0.01% and 0.015%) as treatment groups, Using normal YPD liquid medium as a control, after culturing at 28°C for 24 hours, measure the number of colonies on the YPD agar plate, and finally screen out 8 strains that can tolerate pH = 4.0, 6% sodium chloride concentration and 0.015% sodium nitrite concentration of yeast.
[0034] The acid resistance, salt resistance and nitrite resistance of Debaria hansenii SH4 are as follows: figure 2 , 3 , 4 shown. After the fermentation, the pH of the air-dried sausage is usually between 4.0 and 5.0, and the growth of acid-sensitive yeast will...
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