Application of bacillus mojavensis KJS-3 to decomposition of phenol

[0015] Example 1 Isolation of strains

[0016] The part of the sticky substance in the food waste that cannot be grown by molds is collected and moved to the laboratory, and cultured in a shaker at 37°C for 16 hours in Trypticase Soy Agar (TSA). Here, select the grown The colonies are each inoculated in Trypticase Soy Broth (TSB) for storage. The grown strains are stored at 45°C and 20°C for cultivation, and then the strains grown in this culture are selected for storage before storage . These selected strains were stored at 4°C for 30 days, and then this was cultured again at 37°C in Trypticase Soy Agar Liquid Medium (TSA). In 10 kinds of culture solutions at 37°C Two of the strains with endospore formation within 7 days were selected, and the strains were named No. 3 and No. 4 respectively.

[0017] These strains were entrusted to the Korea Collection of Microorganisms to carry out genetic material analysis and obtained Bacillus mojavensis KJS-3 (Bacillus mojavensis KJS-3) and Bacillus mojavensis KJS-4 (Bacillus mojavensis KJS-4) strains, and the strains were tested. Strain registration. Bacillus mojavensis KJS-3 (Bacillus mojavensis KJS-3) is deposited at the Korea Collection of Microorganisms Bacillus mojavensis KJS-3 (deposit number: KCCM10961P).

[0018] Example 2 Confirmation of phenol decomposition ability

[0019] The above-mentioned two microorganisms, Bacillus mojavensis KJS-3 (Bacillus mojavensis KJS-3) and Bacillus mojavensis KJS-4 (Bacillus mojavensis KJS-4) at a temperature of 40 ℃, tryptone soybean agar medium (TSA ) Inoculated and cultivated. This experiment was performed using Bacillus mojavensis KJS-3 (Bacillus mojavensis KJS-3) cultured at 40°C.

[0020] 1. Inoculate the KJS-3 strain of Bacillus mojave in 10ml TSB medium, cultivate at 40°C for more than 16 hours.

[0021] After culturing for 16 hours, take 5ml out of the culture solution, inoculate it in 100ml TSB, incubate for 90 minutes at 40°C, and keep the stirring speed at 100-200rpm.

[0022] 2. Take 500 μl (μl) of the culture cultured in the above steps, centrifuge at 10,000 rpm for 1 minute and remove the supernatant, then inject 500 μl of normal saline (0.9% NaCl), mix thoroughly, and inoculate in Minimal medium.

[0023] 3. According to different time, after 0min, 30min, 60min, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours and 8 hours, measure the absorbance (OD) value in the UV-visible spectrophotometer to confirm The growth curve of the strain.

[0024] The composition of the minimum medium is as follows: (1L, pH7.0) NH 4 Cl 1.0g, K 2 HPO 4 4.35g, NaH 2 PO 4 3.9g, MgSO 4.7H 2 O 0.48g CaCl 2 0.03g, FeSO 4 0.01g, MnCl 2 0.01g, CoCl 2 0.001g, Na 2 MnO 4 0.001g.

[0025] Prepare 3 minimum culture media, respectively make and test with phenol concentration of 50ppm, 250ppm, and 500ppm. The absorbance is measured at 600nm wavelength as follows:

[0026] 50ppm phenol 250ppm phenol 500ppm phenol 0min 0.032 0.032 0.032 30min 0.057 0.07 0.066 1hr 0.074 0.076 0.094 2hrs 0.114 0.130 0.161 3hrs 0.150 0.181 0.193 4hrs 0.312 0.331 0.317 5hrs 0.519 0.524 0.539 6hrs 0.721 0.687 0.688 7hrs 0.754 0.704 0.735 8hrs 0.731 0.705 0.691

[0027] From the above results, it can be seen that the Bacillus mojave strain KJS-3 uses phenol as the carbon element to decompose and utilize, and it shows the maximum growth state after 7 hours. figure 1.

[0028] Example 3

[0029] For the determination of the phenol content of the phenol-decomposing strain, the reaction experiment was carried out in the following order, and the absorbance was measured at a wavelength of 650nm.

[0030] 1) After the strain is cultured for 8 hours, transfer the culture solution to the refrigerator for storage.

[0031] 2) Phenol quantification

[0032] (1) Standard preparation: 250ppm-phenol, 25ppm-phenol, 12.5ppm-phenol.

[0033] (2) Sample: 500ppm-phenol, containing a small amount of culture medium (cultured at 40°C for 8 hours);

[0034] 250ppm-phenol contains a small amount of medium, and 50ppm-phenol contains a small amount of medium (cultivation at 40°C for 8 hours).

[0035] (3) Experimental process: Take 100ml each for the standard and each sample, and add 6ml of 8wt% K 3 Fe(CN) 6 , 6ml of 0.1M FeCl 3 (0.1M HCl as solvent), after reacting for 5 minutes, measure the absorbance (OD) at a wavelength of 650 nm.

[0036] The results are shown in the following table:

[0037]

[0038] It can be seen from the above table that the OD value of the sample 500ppm is 1.304, about 17ppm of phenol is detected, indicating that 483ppm of the 500ppm phenol is decomposed, and the OD value of the sample 250ppm is 1.108, and about 14ppm of phenol is detected. It shows that 236ppm of phenol in 250ppm is decomposed. Finally, the OD value in 50ppm of the sample is 1.084, which means that about 13ppm of phenol is detected and 37ppm of phenol is decomposed. Therefore, in the present invention, Bacillus mojave KJS- The outstanding characteristics of the 3 strains against phenolic compounds have been confirmed.