Polystyrene pretreatment and enzymatic degradation method
A polystyrene, pretreatment technology, applied in the field of enzyme engineering, can solve the problems of poor accessibility of enzymes, incomplete UV pre-oxidation, low degree of UV pre-oxidation of PS plastics, etc.
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Embodiment 1
[0064] Example 1 Preparation of dye decolorizing peroxidase AnaPX enzyme solution
[0065] The recombinant expression method of the dye decolorizing peroxidase AnaPX is as follows:
[0066] The AnaPX encoding gene (as shown in SEQ ID NO. 2) was connected to the pET-24a(+) vector, and then transferred to the E.coliBL21(DE3) expression host to obtain the recombinant expression strain E.coli BL21(DE3) / pET-24a (+)-AnaPX. The above recombinant expression strains were inoculated into LB medium containing kanamycin resistance, cultured at 200 rpm and 37°C for 10 h; then inoculated into TB liquid containing kanamycin resistance at a ratio of 5% (v / v) medium, cultured at 37°C, when the OD 600 When reaching 0.6, isopropyl-1-thio-β-d-galacturonic acid and 5-aminolevulinic acid were added to a final concentration of 0.4 mM, and then cultured at 30° C. for 24 h. The obtained fermentation broth was centrifuged at 8000 rpm and 4 °C for 20 min, the supernatant was removed, the cells were c...
Embodiment 2
[0071] Example 2: Oxidative pretreatment of low molecular weight polystyrene powder with potassium permanganate and concentrated sulfuric acid
[0072] Weigh 1g PS powder (M w 35000) and 1g potassium permanganate into a 100mL conical flask, first add 10mL ethyl acetate and stir to dissolve PS powder, then add 10mL deionized water to dissolve potassium permanganate, then add 5mL concentrated sulfuric acid, at 55 ℃ Catalyze for 12h, ventilate and volatilize overnight, remove the liquid part by filtration, then rinse with water several times to remove the residual oxidant, and finally place the sample in an oven to dry to constant weight, which is the obtained sample.
[0073] Detect the infrared spectrum of polystyrene (PS) pre-oxidized by potassium permanganate / concentrated sulfuric acid ( figure 1 ), after dissolving in ethyl acetate, potassium permanganate and concentrated sulfuric acid can oxidize polystyrene to a certain extent, and at 1700 cm -1 Oxygen-containing functio...
Embodiment 3
[0074] Example 3: Enzymatic degradation of potassium permanganate / concentrated sulfuric acid pre-oxidized polystyrene by dye decolorization peroxidase in aqueous system
[0075] The polystyrene (PS) after oxidative pretreatment in Example 2 was subjected to enzymatic degradation in the aqueous system, and the AnaPX enzyme solution prepared in Example 1 was adjusted to the optimum pH 3.0 with glycine-hydrochloric acid buffer (100 mM, pH 2.0), and said Take 20mg of chemically pre-oxidized PS powder and add it to 100mL, the enzyme activity is 3.0×10 4 U / L AnaPX enzyme solution (150U AnaPX酶 / mg 聚苯乙烯 ), then add 1 mL of 10 mM H 2 O 2 , at 25°C and 200rpm for 48h. After the reaction, the enzymatic hydrolysis solution of the aqueous phase system was filtered to retain the polystyrene degradation products, first washed with SDS, rinsed with deionized water, then washed with 75% ethanol, and rinsed with deionized water in an oven. After drying in medium, the product was subjected ...
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