Pichia kluyveri var.kluyveri strain, culture method and application thereof
A culturing method and strain technology, applied in the field of microorganisms, can solve the problems of high energy consumption, complex technical procedures, etc., and achieve the effects of low alcohol content, alleviation of environmental pollution, and low cost and energy consumption.
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Embodiment 1
[0035] A strain SS1-5 of Pichia kluyveri var.kluyveri strain SS1-5, which was preserved in the General Microbiology Center of China Committee for the Collection of Microorganisms on December 20, 2010, address: Chaoyang District, Beijing Institute of Microbiology, Chinese Academy of Sciences, No. 1, Beichen West Road, No. 4494.
[0036] The above-mentioned Pichia kluyveri var. kluyveri strain SS1-5 is isolated from freshly squeezed fresh grape juice. The edges and centers of the colonies are cream-colored, with spherical protrusions, rough surface wrinkles, and opaque , not reflective, irregular edges, fuzzy, dull white; cells are oval. After 2-3 days of YEPD liquid culture, precipitation and chorion appeared. The strain is temperature-resistant to 45°C, resistant to pH 2.5-12.0, and resistant to SO 2 400mg / L, resistant to ethanol 8% (vol), resistant to NaCl 100g / L, resistant to glucose 500g / L.
Embodiment 2
[0038] The cultivation method of Pichia kluyveri var.kluyveri strain SS1-5 described in Example 1, the steps are as follows:
[0039] (1) Take Pichia kluyveri var. kluyveromyces strain SS1-5 with an inoculation needle, inoculate it on YEPD solid medium by streaking, and activate the culture at 28°C for 40-48 hours to obtain the activated strain;
[0040] (2) Inoculate the activated strain obtained in step (1) into YEPD liquid medium, and culture it on a shaker at 120 r / min for 18-22 hours at 28°C to obtain a seed solution;
[0041] (3) Inoculate the seed solution prepared in step (2) into the wort liquid culture medium at a volume ratio of 5%, and culture it for 34-36 hours at 25°C to obtain the seed solution for production.
[0042] The components per liter of the YEPD solid medium in the step (1) are as follows: 20 grams of glucose, 10 grams of yeast powder, 10 grams of peptone, 20 grams of agar, distilled water to 1000 milliliters, natural pH, extinguished at 115 ° C Bacte...
Embodiment 3
[0046] The application of the above-mentioned Pichia kluyveri var. kluyveri strain SS1-5 in the preparation of low-alcohol beverages, the steps are as follows:
[0047] (i) The raw barley malt is fed into water at a weight ratio of 1:5 at 52°C and kept for 40 minutes; the temperature is raised to 65°C and kept for 60 minutes; then the temperature is raised to 72°C and kept for 15 minutes for iodine test; 78°C is kept for 10 minutes; After complete saccharification, wash the grains twice, the water temperature for washing grains is 78°C; the boiling time is 70 minutes, and the boiling strength is not less than 8%. Add three times, let it stand for 30 minutes, separate the hops grains and thermal coagulation, and cool to obtain wort with a sugar content of 12±0.3°P;
[0048] (ii) Add the seed liquid for production of Pichia kluyveri var.kluyveri strain SS1-5 to the wort prepared in step (1) according to the volume ratio of 5% , fermented and cultured for 8 days under the condit...
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