A strain of achromobacter xylosoxidans producing carboxymethylcellulase
A technology for carboxymethyl cellulose and cellulose degradation, which is applied in the field of bioengineering and can solve problems such as the lack of alkaline cellulase production strains
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Embodiment 1
[0025] Example 1 strain screening, isolation and identification
[0026] Step 1: Sampling
[0027] Compost samples in the suburbs of Wuming County, Nanning City, Guangxi Zhuang Autonomous Region.
[0028] The second step: primary screening, secondary screening and purification
[0029] Take 1g of the sample, add 10ml of sterile distilled water, shake fully, stand still for 30 minutes, take the supernatant and dilute 105 times, smear and inoculate the CMC-Congo red plate screening medium, culture at 30°C for 72h until the colony grows, pick a transparent Colonies with larger hydrolysis circles were inoculated with CMC liquid fermentation medium, and cultured in shake flasks at 37° C. and 200 rpm for 24 hours, and the alkaline CMC enzyme activity of the culture medium was measured. Select the culture bottle with the highest activity, take the bacterial liquid and inoculate it with CMC-Congo red plate screening medium for culture, purify the strain 3 times, and finally obtain a h...
Embodiment 2
[0035] Example 2 Detection of Cellulase Levels Produced by Bacterial Strains
[0036] according to figure 1 and figure 2 Optimum temperature and pH of growth shown, strain 7B is inoculated with CMC fermentation medium and cultured, and the enzyme production curve is drawn, and the results are as follows: Figure 4 shown. The preparation method of the crude enzyme solution is as follows: the activated bacterial strain is inoculated with the CMC fermentation medium, cultivated at 30°C and 200rpm for 48h, takes the culture solution, centrifuges at 4000rpm for 10min at 4°C, takes the supernatant, and adds 55% (NH 4 ) 2 SO 4 The precipitate was centrifuged at 10,000 rpm for 10 min at 4°C to collect the precipitate, and the same volume of culture medium was added to the pH 8.0, 0.1 mol / L Tris-HCl buffer to dissolve. CMC sodium salt was used as a substrate to measure the activity of endocellulase, that is, CMC enzyme activity. The enzyme reaction system contains 0.5 ml of crud...
Embodiment 3
[0040] Embodiment 3 is to the processing of BTEX and refractory chelating agent
[0041] This example is carried out at laboratory level. A 10-fold diluted LB medium with 2% CMC cellulose was used to simulate the sewage water body rich in organic matter. Respectively: add 0.02% ethylbenzene to simulate BTEX polluted industrial wastewater; add 0.1% IDS (iminodisuccinic acid) to simulate refractory chelating agent polluted water; add 0.1% endosulfan to simulate refractory pesticide polluted water. In the experiment, 100 mL of culture solution was used, the inoculum size was 1%, and the shake flask was cultured at 30° C. with a rotation speed of 180 rpm for 48 hours. Sampling and detection of residual COD and pollutants every 4 hours. Among them, the determination of residual ethylbenzene adopts n-hexane extraction + gas chromatography analysis method, in which the chromatographic column is a HP-Innoax capillary column, the column temperature is 90°C, the column flow rate is 1m...
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