Bacillus amyloliquefaciens for efficiently purifying breeding wastewater and application of bacillus amyloliquefaciens
A technology for dissolving starch spores and aquaculture sewage, which is applied in the field of microbial technology and aquaculture water environment improvement, can solve problems such as poisoning aquaculture organisms, deterioration of aquatic product quality, and decline in water self-purification ability, so as to increase benefits, reduce usage, and reduce water pollution. Effect
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Embodiment 1
[0020] Example 1 Screening and Identification of Bacterial Strains by Protease Activity
[0021] 1. Screening
[0022] (1) Take 50mL of mud samples from the sewage ditch of aquaculture farmers in Weibei Liuchang, Weifang City, bathe in 80°C water for 15 minutes to kill non-spore bacteria; after dilution, spread the samples on nutrient agar medium (peptone 10g, beef Cream 3g, sodium chloride 5g, agar 15-20g, distilled water 1000mL, pH 7.2-7.4) plate, 37°C, culture for 24h; select typical Bacillus colony to streak on the nutrient agar medium plate, 37°C, culture for 24h.
[0023] (2) Primary screening of protease-producing strains: pick a small number of colonies and inoculate them into seed culture solution (10g of peptone, 5g of yeast powder, 10g of sodium chloride, 1000mL of distilled water, pH 7.2-7.4), at a constant temperature of 35°C and 180r / min Cultivate for 24 hours, and dilute the proliferating bacteria suspension into different gradients according to the 10-fold dil...
Embodiment 2
[0046] The evaluation of embodiment 2 Bacillus amyloliquefaciens enzyme production conditions and enzyme activity quality
[0047] 1. The growth curve and enzyme production curve of the strain
[0048] Strain Am11 was inoculated in liquid medium (peptone 10g, yeast powder 5g, sodium chloride 10g, distilled water 1000mL, pH 7.2-7.4), and cultured at 35°C and 180r / min. Sampling every 2h, 6 times in a row, and every 12h, sampling for a total of 72h, measuring OD 600nm To draw the growth curve of the strain, at the same time, centrifuge the bacterial liquid sampled each time at 10 000r / min, take the supernatant to measure the protease activity, and draw the enzyme production curve to determine the maximum enzyme production, maximum enzyme production time and enzyme production of the strain. relationship over time.
[0049] 2. Factors affecting the ability of strains to produce protease
[0050] (1) Preparation of seed liquid: in the ultra-clean bench, use an inoculation loop to...
Embodiment 3
[0058] The preparation of embodiment 3 bacillus amyloliquefaciens microbial preparation
[0059] 1. Preparation of slant strains: Streak inoculation of the above-mentioned Bacillus amyloliquefaciens Am11 on the nutrient agar medium, culture at 37°C for 12 hours;
[0060] 2. Preparation of solid seeds: inoculate the slant strains prepared in step 1 into the fresh nutrient agar slant of the eggplant bottle (10g of peptone, 3g of beef extract, 5g of sodium chloride, 15-20g of agar, 1000mL of distilled water, pH 7.2-7.4), 37 ℃, incubate for 12h;
[0061]3. Preparation of liquid seeds: Wash the solid seeds prepared in step 2 from the slope of the eggplant bottle and inoculate them into a 250L seed tank with an inoculation amount of 0.5%, at 37°C, and cultivate for 12 hours. The components of the medium in the seed tank and their weight The ratios are: bean cake powder 2.5%, corn steep liquor 2%, disodium hydrogen phosphate 0.1%, ammonium sulfate 0.05%, manganese sulfate 0.05%, and...
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