Halogenated benzoic acid degradation strain, inoculant produced by strain and application
A technology of halogenated benzoic acid and degrading bacteria, which is applied in the field of biotechnology, can solve problems such as threats to the environment and human health, and achieve the effects of maintaining human health, low production and use costs, and good repair effects
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Embodiment 1
[0026] Embodiment 1, isolation and identification of bacterial strain
[0027] The present invention provides a kind of bacterial strain capable of efficiently degrading halogenated benzoic acid and the bacterial agent produced thereof, the bacterial strain used is Gram-negative bacteria H8, which is isolated from the soil of a certain pesticide factory in Suzhou, Jiangsu. The specific isolation and screening methods for strains are as follows:
[0028] Take 5.0 g of the soil sample and add it to 100 ml of inorganic salt (hereinafter referred to as MM) medium containing 0.2 mM 3,5-dibromo-4-hydroxybenzoic acid (hereinafter referred to as MM) culture medium at 30 ° C and 150 rpm for 5 days, with 5% inoculum size (v / v) transfer to the same fresh medium, and enrich and culture continuously for four times. Dilute the fifth-generation enrichment solution on MM solid medium containing 0.2mM 3,5-dibromo-4-hydroxybenzoic acid, culture at 30°C for 4 days, pick a single colony on the ...
Embodiment 2
[0030] Embodiment 2, laboratory degradation experiment
[0031] 2.1 Growth utilization and degradation of 3,5-dibromo-4-hydroxybenzoic acid by strain H8
[0032] Detection of 3,5-dibromo-4-hydroxybenzoic acid by high-performance liquid chromatography: take 1mL sample, centrifuge at 12000rpm for 5min, carefully absorb the supernatant, and filter the supernatant through a 0.22μm aqueous membrane filter for detection by HPLC. Detection conditions: Shimadzu RID-10A is the high-performance liquid chromatograph; the chromatographic column is a C18 reversed-phase column, the specification is 250mm×4.6mm; the column temperature is 40°C; the mobile phase is acetonitrile:water:acetic acid (50:50:0.5, V :V:V), the flow rate is 1.0mL / min; the detection wavelengths are 221nm and 250nm.
[0033] The strain H8 was adjusted to a final concentration of 0.06-0.08 (OD 600The inoculum amount) was connected to 100mL MM containing 0.2mM 3,5-dibromo-4-hydroxybenzoic acid, cultured on a shaker at 3...
Embodiment 3
[0045] Embodiment 3, soil degradation experiment
[0046] The vegetable garden soil was taken as the test soil sample. Pass the soil sample through a 2mm sieve, and take a certain amount of 3,5-dibromo-4-hydroxybenzoic acid, 3-bromo-4-hydroxybenzoic acid, 3,5-dichloro-4-hydroxybenzoic acid and 3- Chloro-4-hydroxybenzoic acid powder was dissolved in 100mL of methanol, and then soaked in diatomaceous earth, so that the pesticide was completely absorbed. Diatomite after soaking is placed in fume hood to dry, and it is mixed in the soil, so that the concentration of pesticide in the soil is about 50mg / kg. Take 500g of each soil sample, insert the seed solution according to 10% inoculum amount, and cultivate it in a constant temperature incubator at 30°C. The soil sample with the same amount of sterile MM liquid is used as a control, and the water holding capacity of the soil is maintained at 60%. Samples were taken at 0, 1, 3 and 5 days respectively, and 3 samples were taken fo...
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