Rhodosporidium paludigenum for degrading gossypol and application thereof
A technology of Rhodosporidium spore and free gossypol, which is applied in the field of microorganisms, can solve the problems of shortage of protein feed resources, low feeding rate of cottonseed meal, unstable performance of gossypol degrading bacteria, etc., so as to shorten the start-up time and achieve good removal effect. , The effect of low production and use cost
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Embodiment 1
[0019] Embodiment 1 strain mutagenesis and preliminary screening
[0020] 1) Rejuvenation of strains
[0021] Remove the liquid paraffin from the strain Rhodotorula HJ-Y obtained in the original test and stored at 4°C, put 1 mL of its bacterial suspension into a 250 mL Erlenmeyer flask with 100 mL of PDA liquid medium, and shake it in a constant temperature steam bath at 28°C. The bed was cultivated for 48 hours, and the rotation speed was 200r / min.
[0022] The screening method of the yeast HJ-Y is the same as that of Xia Xincheng, Li Guiqiang, Teng Anguo, etc. Screening and identification of strains for compound fermentation of cottonseed meal and optimization of fermentation process parameters, [J] Journal of Chinese Cereals and Oils, 2010, 25(1): 91-97 strain screening method.
[0023] 2) Strain selection
[0024] On the ultra-clean operating table, use the inoculation loop to pick up the rejuvenated bacterial liquid on the PDA solid medium plate for subculture, and sel...
Embodiment 2
[0041] Embodiment 2 Bacterial classification re-screening
[0042] The original species of HJ-D-20, HJ-D-30, HJ-Z-20-D-20 and the original species of control strain HJ-Y screened in Example 1 were respectively activated on the screening medium, and the activated The bacterial strains were inoculated on the slant of the test tube (see the screening medium for the formula) for later use. The test tube species were respectively inoculated in 1000ml shake flasks containing 200ml potato liquid medium, cultured with constant temperature shaking until the logarithmic phase, and the activated strains were inserted into 20ml potato liquid culture to obtain the first-grade strains based on the 100ml Erlenmeyer flask (the condition was 30 ℃, 200r / min shaker culture for 48h), take the first-grade strain and insert 1ml into a 250ml Erlenmeyer flask equipped with 100ml potato liquid medium as the second-grade strain (conditions are 30°C, 200r / min shaker culture for 48h ).
[0043] Use HJ-...
Embodiment 3
[0048] 1) Inject Rhodosporidium yeast HJ-D-20 (CGMCC No.4372) into 20ml potato liquid culture to obtain the first-grade strain based on a 100ml Erlenmeyer flask (conditions are 30°C, 200r / min shaker culture for 48h), take one Inject 1 ml of the first-grade strain into a 250-ml Erlenmeyer flask equipped with 100 ml of potato liquid medium to be the second-grade strain (conditions are 30° C., 200 r / min shaker culture for 48 h).
[0049] 2) The secondary bacterial strains cultivated to the logarithmic phase were respectively inserted into the cottonseed meal fermentation substrate with a free gossypol content of 614mg / kg, the inoculum size was 5%, placed in a 500ml Erlenmeyer flask, 30°C, constant temperature and static Place and culture for 72h. Dry, measure FG and CP content, analyze fermentation effect, see Table 3. It can be seen from Table 3 that after fermentation by strain HJ-D-20, the degradation rate of FG reached 90.80%, the increase rate of crude protein was 24.59%, a...
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