A high copper-resistant and high iron-resistant Leptospira siderophilus byl and its application
A technology of Leptospira ferrous and copper resistance, applied in the field of microorganisms, can solve the problem of not showing high-concentration iron ion and copper ion tolerance, not showing high-concentration copper ion and iron ion tolerance, affecting metal leaching And other issues
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Embodiment 1
[0019] Embodiment 1 Isolation and identification of bacterial strains of the present invention
[0020] (1) Take 1ml of water sample from the pit water of Baiyin deep copper mine and inoculate it to contain 20g / L FeSO 4 ·7H 2 O, in 9k medium with pH 2.5-2.8, 30°C enrichment culture for 5 days. Inoculate the bacterial solution obtained from the above-mentioned enrichment culture to a concentration of 60g / L FeSO 4 ·7H 2 O, in 9k medium with pH 2.5-2.8, cultured at 40°C, pH 1.8, passaged once every 3 days, to obtain a bacterial liquid containing Leptospira. Then transfer the above bacterial solution to pH1.6 containing 60g / L FeSO 4 ·7H 2 O, 28g / L ferric sulfate, cultivated in the 9k subculture medium of pH2.5-2.8 for 3 days, and serially diluted and passaged for 3 times (passage gradient is 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 ), and then transfer the last grown bacteria to the 9k subculture medium with pH 0.8, and then transfer to the 9k sub...
Embodiment 2
[0026] Embodiment 2 Growth characteristics of bacterial strains of the present invention
[0027](1) Take substratum 2 as bacterial growth substratum, add 50ml substratum in 250ml Erlenmeyer flask, the inoculum size of Leptospira ferrophilum BYL is 10%, adjusting pH is 1.0, 1.2, 1.4, 1.6, 1.8, 2.0; 180rpm, cultured at 40°C for 36h, and the iron oxidation rate was determined by potassium dichromate titration. Iron Oxidation Rate IOR (Iron Oxidation Rate) as figure 2 As shown, the optimum pH is 1.4.
[0028] (2) Take culture medium 2 as bacterial growth medium, add 50ml culture medium in 250ml Erlenmeyer flask, the inoculum size of Leptospira ferrophilum BYL is 10%, adjust pH1.4, at temperature be 30 ℃, 35 ℃ , 40°C, 45°C, and measure the iron oxidation rate. The result is as image 3 As shown, the optimum growth temperature range of the bacteria is 35-40°C, and it has oxidation ability at 45°C.
Embodiment 3
[0029] Example 3 Oxidation and tolerance of bacterial strains of the present invention to high-concentration iron ions
[0030] Change the concentration of ferrous sulfate in medium 2 to the following gradient (g / L): 10, 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, the pH value was adjusted to 1.3, and 10 mL of medium with different concentration gradients was used to inoculate Leptospira ferrophilum BYL according to the inoculum size of 10% (v / v), and cultured with shaking at 40°C. After 36 h of oxidation, the concentration of ferrous ions in the culture was measured. get as Figure 4 Shown result, when growing in the culture medium that ferrous sulfate concentration range is 10-220g / L, along with the raising of iron ion concentration, iron oxidation efficiency also increases thereupon; When growing in the culture medium of / L, with the increase of iron ion concentration, the iron oxidation efficiency is obviously reduced. ...
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