Acinetobacter calcoaceticus and culture method and application thereof
A cultivation method, calcium acetate technology, applied in the field of microorganisms, can solve the problems of slow degradation of petroleum, secondary pollution, limited number of viable bacteria, etc., achieve obvious effects, speed up the restoration process, and improve the effect of the micro-ecological environment
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[0034] The present invention will be further described below in conjunction with the examples, but the protection scope of the present invention is not limited thereto.
[0035] Acinetobacter calcoaceticus (Acinetobacter calcoaceticus) WG-6 described in the examples was deposited in the General Microbiology Center of China Committee for the Collection of Microbial Cultures on June 11, 2010, with a preservation number of CGMCC No.3915.
[0036] The peat described in the examples was purchased from Guanghe Garden Group, the bran was purchased from Dongjia Flour Mill in Licheng District, Jinan City, and the crude oil was from the dehydrated crude oil in Gudong Oilfield, Shengli Oilfield. Other reagents used in the examples were commercially available common products.
Embodiment 1
[0037] Embodiment 1: the degradation of petroleum by Acinetobacter calcoacetate
[0038] Acinetobacter calcoaceticus (Acinetobacter calcoaceticus) WG-6 was picked and inoculated on beef extract peptone slant medium, and activated for 16 hours.
[0039] The components of the beef extract peptone slant medium are as follows: 3 g beef extract, 5 g peptone, 5 g NaCl, 1.5 g agar, 1000 mL distilled water, pH 7.2.
[0040] Pick Acinetobacter calcoaceticus from the beef extract peptone slant medium and connect it to the liquid beef extract peptone medium, cultivate it on a shaker with a rotation speed of 250rpm at 28°C for 24 hours, and then inoculate it into crude oil culture according to the volume ratio of 5% inoculum Medium, cultured on a shaker at 28°C for 7 days with a rotation speed of 150rpm. The crude oil culture medium without bacteria was used as blank control. At the end of the cultivation, add chloroform to the medium for extraction, heat the extract to 75°C to volatilize...
Embodiment 2
[0047] Embodiment 2: the preparation of the high-density microbial agent that can remediate petroleum-contaminated soil
[0048] Acinetobacter calcoaceticus (Acinetobacter calcoaceticus) WG-6 culture:
[0049] (1) Inoculate the Acinetobacter calcoaceticus (Acinetobacter calcoaceticus) WG-6 with the strain preservation number of CGMCC No.3915 on the beef extract peptone slant medium (the components are the same as in Example 1), and the culture temperature is 28°C , activated for 24 hours, and then transferred to liquid beef extract peptone medium at a culture temperature of 20°C and activated for 16 hours in a shaker flask to obtain the activated bacterial liquid;
[0050] (2) the bacterium liquid after the activation that step (1) is made is transferred in the liquid fermentation medium in the ratio of 5% by volume, shaker cultivates 16h, and rotating speed is 200rpm, and cultivation temperature is 20 ℃, obtains calcium acetate not Acinetobacter bacteria liquid. After testi...
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