(Pichia jadinii)Hzd-pj-003 and application in fermentation of feed
A technology for Pichia pastoris and fermented feed, which is applied in the field of Pichia pastoris to achieve the effects of increasing protein content and hydrolytic enzymes, strong acid resistance, and improving feed utilization.
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Embodiment 1
[0013] Obtaining Pichia jadinii Hzd-pj-003 and its culture condition test
[0014] Screening and Identification of Pichia jadinii Hzd-pj-003
[0015] 1) Screening of Pichia jadinii Hzd-pj-003
[0016] From the old peanut shells piled up by farmers in Nancun Township, Pingdu, Shandong Province, select the ones with some signs of fermentation on the surface as samples, soak them in sterile water, and use the separation method of gradient dilution and coating to dilute the peanut shell solution by 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Afterwards, draw 0.1 mL each and apply it on the PDA medium to separate the strains, culture them in a constant temperature incubator at 28°C for 2-3 days, and pick the colony on the PDA medium that is milky white, smooth, and the diameter of the colony is about 3-5mm. Single colonies were repeatedly streaked until purified, such as figure 1 shown.
[0017] The formula of the PDA medium is as follows: 200 g of potatoes, 20 g of glucose, 15-20 g o...
Embodiment 2
[0034] Acid Resistance Test of Yeast Hzd-pj-003
[0035] Preparation of Pichia pastoris strain: Inoculate the preserved strains into 250mL eggplant bottles filled with 70-80mL solid slant PDA medium, and cultivate them in a constant temperature incubator at 30°C for 3 days; Rinse down with sterile water, and repeatedly blow and suck to mix the bacteria and sterile water evenly to obtain a bacterial suspension, insert 3% of the inoculum into a 500mL triangular flask containing 300mL YPD medium, and culture on a constant temperature shaker at 30°C 24h, rotating speed 220rpm; according to 3% inoculum amount, put it into a 500mL Erlenmeyer flask containing 300mL fermentation medium, cultivate in a constant temperature shaker at 30°C for 24h, rotating speed 220rpm; measure the OD of the fermentation medium every 4h 660 values, and the results are shown in Table 2. The test results show that the yeast Hzd-pj-003 can still grow slowly when the pH value is 2, and has strong acid resi...
Embodiment 3
[0043] Pichia jadinii Hzd-pj-003 applied to fermented feed
[0044] Pick a single colony from the slant culture medium and put it in a 500ml Erlenmeyer flask with 300ml seed medium, shake it at 30°C and 220r / min, when the OD in the liquid seed medium 660When the value reaches 1.4, the inoculum amount of 3% to 5% is inserted into a 500ml Erlenmeyer flask containing 300ml fermentation medium under sterile conditions, and shaken at 30°C and 220r / min for 24h, that is, the preparation of Bacillus subtilis Hzd- bs-001 fermentation broth.
[0045] The formulation of the seed medium (YPD medium) is as follows: yeast extract 5g / L, peptone 10.0g / L, glucose 20.0g / L, sodium propionate 2.5g / L, pH 6.8. Sterilize at 115°C for 30min.
[0046] The formula of the fermentation medium is as follows: 6% glucose, 1% corn steep liquor, 0.25% yeast powder, 0.5% soybean peptone, KH 2 PO 4 0.1%, MgSO 4 0.1%, pH 6.0-6.2, sterilized at 121 ℃ for 20min.
[0047] Proportion of fermented feed materia...
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