Aroma producing yeast and application thereof in fermented soybean milk
A technology for producing aroma yeast and fermenting soymilk, which is applied in the field of yeast and fermented soymilk processing, can solve the problems such as difficult to remove beany smell, achieve the effect of improving flavor characteristics, simple cultivation conditions, and good development and application prospects
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[0031] (4) Preparation of lactic acid bacteria and yeast seed liquid:
[0032] ①Lactobacillus XPL-1 seed solution: Take a 100mL Erlenmeyer flask, add 50ml of 12% (w / v) milk medium, bathe in water at 60°C for 30min, then sterilize at 115°C for 15min. Inoculate XPL-1 bacterial powder at a ratio of 0.004g / 100ml, incubate at 37°C for 12 hours, and use it as a seed culture solution for later use.
[0033] ② Aroma-producing yeast PichiaamethioninaY seed liquid: take a 100mL Erlenmeyer flask, add 50mL wort liquid medium, sterilize at 115°C for 15min, inoculate PichiaamethioninaY strain, and culture at 36°C for 24h as a mother starter. Take a 100mL Erlenmeyer flask, add 40ml of 12% (w / v) milk medium, bathe in water at 60°C for 30min, and then sterilize at 115°C for 15min. The above-mentioned mother starter was added to the milk medium at a ratio of 1 / 40 (v / v), cultured at 36° C. for 24 hours, and used as a seed culture solution for later use.
[0034] ③ Wine active dry yeast VIC see...
Embodiment 1
[0036] Example 1 strain screening
[0037] (1) Using sterile sampling bottles, sample the naturally fermented physalis water on site and bring it back at low temperature. Immediately dilute to 10 with sterile water -3 、10 -4 、10 -5 times, spread on malt juice agar medium. Then put it into a constant temperature incubator at 37°C and cultivate for 48h. Pick suspected colonies and separate them by streaking on plates, repeat this 4-5 times until pure single colonies are obtained. The purified single colony was inoculated on the malt juice agar slant medium with an inoculation needle, and stored in a refrigerator at 4°C.
[0038] (2) isolate bacterial strain Y from the physalis water of natural fermentation, its bacterial colony on wort agar medium is circular, and diameter is generally 3-5mm, milky white, opaque shape, moist sticky, easy to be provoked ( Figure 1a ); Gram staining and cell shape observation were carried out, and the thalli of bacterial strain Y was cylind...
Embodiment 2
[0039] Example 2 strain identification
[0040] Use the DNA extraction kit to extract yeast DNA and perform agarose gel electrophoresis, then design yeast 26SrDNA primers, and perform PCR amplification. The electrophoresis is as follows figure 2 As shown, the band is single, and the band size is about 500bp. (1,2 are PCR amplification products of yeast 26SrDNA sequence; M is 10000bpMarker).
[0041]The purified and recovered DNA fragments were sequenced, and the sequencing results are shown in SEQ.ID.NO1. Then use BLAST software to compare with the reported 26S rDNA, and the comparison results show that the DNA sequence is consistent with Issatchenkiaorientalis, Pichiakudriavzevii and Pichiakudriavzevii in the genus Pichia. The similarity between Pichia amethionina and the like reaches more than 99%. Therefore, it was further inoculated on the YT identification plate, and after culturing at 28°C for 72 hours, the results of YTMroPlate assimilation test (Table 1a) and dissi...
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