Bacillus high temperature bacteria NJAU-N30 for accelerating composting and decomposition and application thereof
A bacillus and composting technology, which is applied in the application field of preparing organic fertilizers, can solve the problems of unpredictable H1-7 bacterial agents, accelerate compost maturity, etc., and achieve the effects of promoting temperature rise, promoting germination index, and promoting growth
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Embodiment 1
[0036] Embodiment 1, isolation and identification of functional strains
[0037] Put the samples in the high temperature phase of composting into a 250ml Erlenmeyer flask with glass beads and 100ml of sterile water, shake at 25°C and 170r / min for 20min to form a soil suspension, and draw 10 -2 、10 -3 、10 -4 、10 -5 Dilute 0.1ml of the soil suspension, spread it on the lignocellulose culture plate, repeat each concentration 3 times, culture at 28°C for 5 days, pick and purify the colonies growing on the plate for more than 5 times, and select the colonies that can still grow Transfer to slant culture and store at 4°C for later use.
[0038]Through qualitative and quantitative screening, a bacterial strain was finally obtained named NJAU-N30. The colony of the bacterial strain NJAU-N30 on the LB plate was small, yellow and shiny, round in shape, slightly raised in the middle, and scattered round in the edge. The bacteria are soft, have a certain viscosity, and are easy to sti...
Embodiment 2
[0047] Determination of various enzyme activities of NJAU-N30 under embodiment 2, high temperature growth conditions
[0048] Use LB medium as NJAU-N30 enzyme production medium: 10g peptone, 5g yeast powder, 10g NaCl, 1000ml deionized water, pH 7.0, sterilized at 121°C for 20min. Use 3ml of sterile water to prepare the bacteria suspension, take 0.5ml and inoculate them in the enzyme-producing medium respectively, and culture on a shaking table at 50°C at 170r / min. After 24 hours, the bacterial liquid was taken, and the extract was added to extract, and the cell membrane was broken to form a stock solution by using an ultrasonic breaker (300W, ultrasonic for 3s, with an interval of 7s, for 3min).
[0049] Cellulase activity was determined according to the method of Albrecht et al. (2008). The stock solution was mixed with acetic acid buffer (50mM, pH 5) at a ratio of 1:5 (w / v), shaken on a horizontal shaker for 1 hour, and then centrifuged at 10000r / min at 4°C for 10 minutes t...
Embodiment 3
[0056] Embodiment 3, the generation of liquid inoculum
[0057] Streak the NJAU-N30 strains stored in glycerol tubes at -80°C on LB solid medium plates for activation, and cultivate them in a 37°C incubator for one hour. Pick a single colony of NJAU-N30 and culture it in a 3ml liquid test tube at 37°C with shaking at 170r / min for 10 hours, and the bacterial solution is used as the seed solution. Transfer the seed solution to the LB liquid shake flask with an inoculum amount of 1% (v / v), cultivate it at 37°C and 170r / min until the mid-logarithmic phase (OD600=1.0), and collect the bacteria by centrifugation at 4°C. Wash with distilled water for 3 times, and resuspend with equal volume of distilled water to obtain liquid inoculum.
[0058] The preparation method of the LB culture medium used is as follows, taking the preparation of 1L medium as an example: protein building 10g, yeast powder 5g, NaCl 10g agar 20g, constant volume to 1000ml, pH natural, sterilized at 121°C for 20...
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