Neosphingosine arerophile, thallus preparation, intracellular enzyme preparation and application of neosphingosine arerophile in degradation of microcystic toxins
A technology for sphingosine and aromatic hydrocarbons, which is applied in the application of bacteria degrading microcystins, new sphingosine aromatic hydrocarbons, and microbial preparations, which can solve the problem of ineffective elimination of extracellular toxins, destruction of algal cytotoxins, and microbial preparations. secondary pollution and other problems, to achieve the effect of improving the micro-ecological balance, fast degradation, and strong survivability
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Embodiment 1
[0078] Example 1 Bacterial preparation and preparation method thereof of Arylophilic neosphingosine strain P6D9
[0079] (1) Prepare liquid culture medium, whose components are ordinary nutrient broth medium (per liter): peptone 10g, beef extract 3g, sodium chloride 5g, pH7.4, or other sources containing carbon and nitrogen to meet the basic growth of microorganisms For the required medium, autoclave at 121°C for 15 minutes;
[0080] (2) Prepare MSM medium for preserving P6D9 bacteria: MgSO 4 ·7H 2 O 0.5g, KH 2 PO 4 0.5g, K 2 HPO 4 0.5g, NaCl 0.5g, FeSO 4 ·7H 2 O 0.01g, CaCl 2 0.02 g, add MC-LR (to make the final concentration of MC-LR in the culture solution about 100 μg / L), adjust the pH value to about 6.5, and sterilize at 121°C for 15 minutes.
[0081] (3) Inoculate the seed liquid of Neosphingoarthophilus P6D9 bacteria (picking a ring of bacteria from the nutrient agar slope or plate of the activated strain and inoculate it in a conical flask containing 30mL BH...
Embodiment 2
[0082] Example 2 Enzyme preparation and preparation method of Arylophilic neosphingobacterium P6D9 strain
[0083] Inoculate the seed liquid of Neosphingoarthophilus P6D9 bacteria (picking a ring of bacteria from the nutrient agar slant or plate of the activated strain and inoculate it in a triangular flask containing 30mL of BHI broth for 20h ± 2h, the obtained liquid Culture) inoculated in the expanded nutrient broth according to 1% inoculum amount, cultivated (18±2) hours at 30°C and 150 rpm, and then centrifuged at a high speed (12000 rpm) for 10 minutes to harvest the bacteria and bacterial culture For the supernatant, wash the cells twice with normal saline solution and resuspend them in normal saline with 10% of the volume of the original bacterial solution, immediately ice-bath, and perform ultrasonic crushing in an ice-bath environment. Ultrasonic crushing conditions: power 300W, interval 10 seconds, ultrasonic oscillation for 6 seconds, and crushing time of 20 minute...
Embodiment 3
[0084] Example 3 Adding MC-LR to simulate the removal effect of microcystins in MC-LR polluted water
[0085] Water samples were collected in Chengbi Lake (located in Nanchang County, Jiangxi Province) where the water quality was good and no cyanobacteria blooms occurred, and no MC-LR was detected in it. Add MC-LR standard substance directly to the two water samples, so that the MC-LR content in the water body is about 260μg / L and 25μg / L respectively, simulating the water body polluted by algae bloom.
[0086] Group (1) the bacterium agent in embodiment 1 is evenly sprinkled in two parts of simulated water bodies, makes the concentration of adding thalline in water about 10 5 bacteria / mL.
[0087] Group (2) The enzyme preparation in Example 2 was added to two simulated water bodies in an amount of 10 mL per liter of water.
[0088] Group (3) the thalline preparation of embodiment 1 and the enzyme preparation of embodiment 2 are added respectively in two parts of simulated wa...
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