Rhodococcus etherivorans ZHC and application thereof in degradation of methyl acrylate
A technology of Rhodococcus etherophora and methyl acrylate, which is applied in the direction of bacteria, microbial-based methods, water/sludge/sewage treatment, etc., can solve the problems of uneconomical and poor effects, and achieve high-efficiency degradation ability and good degradation effect, the effect of broad application prospects
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Embodiment 1
[0033] Example 1: Isolation, purification and identification of Rhodococcus aetherivorans ZHC.
[0034] 1. Isolation and purification of Rhodococcus aetherivorans ZHC.
[0035] (1) Strain screening
[0036] Rhodococcus aetherivorans ZHC is a Gram-positive bacterium domesticated and isolated from activated sludge. The specific steps are as follows:
[0037] Add 50mL of inorganic salt culture solution to a 330mL shake flask, add 10mL of activated sludge (MLVSS 75mg / L) from a sewage plant, then add methyl acrylate with a final concentration of 95mg / L, and enrich at 30°C and 160rpm Cultivate, when the concentration of methyl acrylate is 50% of the initial addition, take out 5mL of the enrichment solution and inoculate it into 50mL of fresh inorganic salt culture solution, add the same amount of methyl acrylate to make the final concentration 95mg / L, repeat the above enrichment After 5 times of process, the last enrichment liquid was diluted 10, 100, and 1000 times with inorganic s...
Embodiment 2
[0053] Embodiment 2, acquisition of resting cells of Rhodococcus etherophore ZHC
[0054] ① Incline cultivation:
[0055] Inoculate Rhodococcus etherophore ZHC into LB liquid medium, culture at 30°C, 160rpm for 2 days, then line the activated bacteria on a solid LB plate in a 30°C incubator, take a single colony and continue to line the plate to detect bacteria The purity of the LB test tube was stored on a slant plane (4°C).
[0056] ② Expanded cultivation
[0057] Inoculate the slant bacteria in step ① into LB liquid medium, cultivate at 30°C and 160rpm for 12 hours to obtain the expanded culture medium, centrifuge at 8000rpm for 10min, collect the wet bacteria, wash with the inorganic salt culture medium, and obtain the static strain of Rhodococcus etherophorum ZHC information cells.
Embodiment 3
[0058] Example 3: Degradation performance detection of different concentrations of methyl acrylate by Rhodococcus etherovorans ZHC.
[0059] Divide the inorganic salt culture solution into 330mL shake flasks each with a volume of 50mL, and sterilize at 110°C for 40min. After the sterilization, it was placed at room temperature for 2 days to confirm that there was no growth of miscellaneous bacteria. Add the quiescent cells obtained by the method of Example 2 with a final concentration of 30 mg / L (in terms of dry cell weight), then add methyl acrylate as the only carbon source to make the final concentrations 95, 190, 285, 380, 475 mg / L respectively, After the shake flask was sealed with a polytetrafluoroethylene stopper, it was cultured on a shaker at 30°C and 160 rpm, and a blank control without adding bacteria was made. Measured by gas chromatography, the detector used is a flame ionization detector, the chromatographic column model is Agilent 19091N-213 (30m × 320um × 0.5u...
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