Keratinolytic microbacterium and its culture method and application
A technology of microbacteria and keratin, applied in the field of environmental microorganisms, can solve problems such as high cost and secondary pollution, and achieve the effects of strong environmental adaptability, high phosphorus removal efficiency and good application prospects
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Embodiment 1
[0022] Example 1 Isolation, screening and identification of high-efficiency phosphorus-accumulating bacteriokeratinolyticum Mk-6 strain
[0023] (1) Medium
[0024] Beef extract peptone solid medium: beef extract 3 g / L, peptone 10 g / L, NaCl 5 g / L, agar 20 g / L, pH 7.0~7.2 (liquid medium without agar), used for the isolation of bacteria ,purification.
[0025] Phosphorus-limited (phosphorus-rich) solid medium: Dissolve 8.372 g of 3-morpholine propanesulfonic acid and 0.717 g of N-tris(hydroxymethyl)methylglycine in 30 mL of deionized water, and adjust the pH to 7.4, add the solution in the following order: 0.01 mL1.830% FeSO 4 solution (prepared), 5 mL 1.9 mol / L NH 4 Cl, 1 mL 0.276 mol / L K 2 SO 4 , 0.025 mL0.02 mol / L CaCl 2 2H 2 O, 0.42 mL 1.25 mol / L MgCl 2 6H 2 O, 20 mL 2.5 mol / L NaCl, 0.02 mL trace element mixed solution (ammonium heptamolybdate 3×10 -6 mol / L, H 3 BO 3 4×10 -4 mol / L, CoCl 2 3×10 -5 mol / L, CuSO 4 10 -5 mol / L, MnCl 2 8×10 -5 mol / L, Zn...
Embodiment 2
[0037] Example 2 Determination of Growth Curve of Mk-6 Strain in Low Nutrient High Phosphorus Synthetic Sewage Culture Solution
[0038] (1) Strain: Mk-6 strain.
[0039] (2) Medium
[0040] Beef extract peptone solid medium: the formula is the same as in Example 1
[0041] Synthetic sewage liquid medium: sodium acetate 0.925 g / L, peptone 0.1 g / L, yeast extract 0.01 g / L, NaCl 0.05 g / L, KH 2 PO 4 0.0655 g / L, MgCl 2 ·6H 2 O 0.1412 g / L, CaCl 2 0.025 g / L, pH 7.0~7.2.
[0042] (3) Method
[0043] Take 20 μL of glycerol bacteria stored at -20°C and spread it on the plate of beef extract peptone solid medium, culture it at 28°C for 24 hours, pick a single colony and mix it with 0.3 mL sterile water, take 100 μL and spread it on beef extract On the plate of peptone solid medium, use a puncher (pore diameter 0.5 cm) to take 2 wells and connect it to 15 mL beef extract peptone liquid medium, and place it in a shaker at 28 °C and 170 r / min for 24 h (OD 600 ≈1.5) was used as th...
Embodiment 3
[0046] Example 3 The effect of liquid loading on the biological phosphorus removal effect of the Mk-6 bacterial strain
[0047] (1) Strain: Mk-6 strain.
[0048] (2) Medium
[0049] Beef extract peptone solid medium: the formula is the same as in Example 1;
[0050] Synthetic sewage liquid culture medium: formula is the same as embodiment 2.
[0051] (3) Method
[0052] Take 20 μL of glycerol bacteria stored at -20°C and spread it on the plate of beef extract peptone solid medium, culture it at 28°C for 24 hours, pick a single colony and mix it with 0.3 mL sterile water, take 100 μL and spread it on beef extract On the plate of peptone solid medium, use a puncher (pore diameter 0.5 cm) to take 2 wells and connect it to 15 mL beef extract peptone liquid medium, and place it in a shaker at 28 °C and 170 r / min for 24 h (OD 600 ≈1.5) was used as the seed solution; and then 8% inoculum volume was inserted into the liquid medium containing different amounts of synthetic sewage (...
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