A selenium-enriched Candida glabrata strain fxy-4 and its cultivation method and application as a fish feed additive
A technology of Candida glabrata and FXY-4 is applied in the field of microorganisms to achieve the effects of enhancing immunity, increasing added value and improving economic benefits
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Embodiment 1
[0020] Candida glabrata FXY-4, which can be obtained as follows:
[0021] Isolation of the original strain:
[0022] Dilute the pickle juice sample and spread it on the plate of YEPD medium, cultivate it at 28°C, observe the growth state of the colony at regular intervals, and pick a single colony and transfer it to the YEPD slant for storage. After preliminary screening, the following yeasts are obtained: Strains: FXY-1, FXY-3, FXY-4, FXY-5, FXY-7, FXY-10, FXY-11, FXY-19, FXY-21, FXY-23, FXY-24, FXY-25, FXY-38, FXY-42, FXY-43, FXY-44, FXY-46, FXY-47, FXY-49, FXY-50, FXY-53, FXY-55, FXY-56.
[0023] containing Na 2 SeO 3 (final concentration is 0.5g / L) in the YEPD substratum that carries out the screening of selenium-enriched yeast, then the yeast that preliminary screening obtains is cultivated with YEPD substratum, and every strain bacterium is divided into blank group and experimental group and cultivates, Add Na to the experimental group 2 SeO 3 (final concentration ...
Embodiment 2
[0025] Sequence Analysis of 18S ITS rRNA Gene of Yeast Strain FXY-4 (Deposit Number: CCTCC NO: M 2015664):
[0026] (1) Extraction of chromosomal DNA (small amount):
[0027] 1. Take 1.5mL bacterial liquid in a 1.5mL Eppendorf tube and centrifuge at 12 000r / min for 5min;
[0028] 2. Discard the supernatant, resuspend the pellet in 900 μL phosphate buffered solution (PBS), and centrifuge at 12 000 r / min at 4 °C for 5 min;
[0029] 3. Discard the supernatant, add 300TE and 200μL 10mg / mL lysozyme, blow and aspirate to mix, incubate at 37°C for 1h, and mix by inverting every 15min;
[0030] 4. Add 600TENS lysate (200mmol / L NaCl, 100mmol / L Tril-HCl pH8.0, 2.0% SDS, 50mmol / L EDTA, 0.5% Triton X-100) and 20mg / mL proteinase K 10μL to the pellet, mix by inversion , incubate at 55°C for 1 hour, and mix by inverting every 15 minutes;
[0031] 5. Centrifuge at 12 000r / min at 5.4°C for 5min, and take the supernatant;
[0032]6. Add an equal volume of P:C:I (25:24:1) to the supernatant,...
Embodiment 3
[0060] Yeast FXY4 (Deposit number: CCTCC NO: M 2015664), Bio Mérieux VITEK 2 automatic identification analysis
[0061] Pick the bacterial lawn of the strain to be tested and inoculate it on the YEPD plate, cultivate it to the logarithmic growth phase at 30°C, dip the Vitek liquid in the kit with a sterilized cotton swab, scrape off the bacterial lawn on the plate, and then dissolve it in 1.8ml Vitek liquid , oscillate and mix well; use a turbidimeter to adjust the turbidity to the specified range (2MacF); then use a pipette gun to add the bacterial suspension to the Vitek identification plate, and the bacterial solution is well filled with the identification hole; then incubate at 30°C for 24 to 48 hours When the identification plate was taken out, the identification plate was read and analyzed with the Bio Mérieux VITEK 2 automatic identification analyzer. The results are shown in Table 3. The physiological and biochemical results are the same as the typical physiological and...
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