Paenibacillus polymyxa strain
A technology of polymyxa spore and strain, which is applied in the field of polymyxa spore strain to achieve the effects of inhibitory activity, significant fresh-keeping effect and outstanding fresh-keeping effect
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Embodiment 1
[0046] Example 1 Bacterial strain isolation and identification
[0047] 1. The inventor collected the orchard soil of Dongguan Agricultural Science Research Center on August 2, 2011, weighed 10g of the mixed soil samples at each sampling point, and put them into 100mL sterile water (prepared 1:10 soil suspension) And in the Erlenmeyer flask with glass beads, shake it for about 20 minutes to make the soil sample and water fully mix and disperse, this is the original dilution; then use a 1mL sterile pipette to draw 1mL of the diluted soil solution into a 9mL sterile In the test tube of water, use the suction ear ball to blow and suck several times to mix well, this is 10 -1 Dilute the suspension; by analogy, dilute the soil sample gradient to 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 The soil dilution suspension was repeated 3 times. The prepared dilution was blown and mixed several times, and 10 -2 、10 -4 、10 -6 Use a sterile pipette to draw three gradient dilutions (breat...
Embodiment 2
[0060] Embodiment 2 antibacterial test
[0061] Since anthracnose and litchi downy mildew are the most common diseases of postharvest litchi, anthracnose and litchi downy mildew were selected as pathogenic indicator bacteria in this example to study the antibacterial activity of Paenibacillus polymyxa strain dgnkzx004.
[0062] 1. Preparation of suspension
[0063] Paenibacillus polymyxa ( Paenibacillus polymyxa ) dgnkzx004 glycerol tube cryopreserved strains were inoculated into 20mL improved LB liquid medium at 2% (v / v), and placed in a 32°C incubator for static activation and culture for 20 hours; the activated seed liquid was pressed 5% (v / v) inoculum was transferred to 1000mL improved LB liquid medium, and cultured in a 32°C incubator with shaking for 20 hours; after the fermentation was completed, the fermentation broth was centrifuged at 8000r / min and 4°C for 15min to collect the bacteria. Wash twice with sterile saline, and centrifuge to obtain the desired viable cel...
Embodiment 3
[0079] Example 3 Inhibition of enzyme activity test
[0080] Since polyphenol oxidase (PPO) and peroxidase (POD) are the two most important enzymes in the browning process of litchi, PPO and POD were selected for enzyme activity inhibition test in this experiment.
[0081] 1. Preparation of crude enzyme solution:
[0082] Take 2g of lychee peel, grind it with 0.2M pH6.8 phosphate buffer solution with 5 times the volume to weight ratio plus 0.4g PVP in an ice bath, centrifuge at 9500r / min at 4°C for 15min, collect the supernatant as the crude enzyme solution, and store at 4°C Save for later.
[0083] 2. Determination of POD enzyme activity
[0084] (1) The reaction system includes: 2175 μL pH6.8 phosphate buffer (composed of: disodium hydrogen phosphate and sodium dihydrogen phosphate, prepared by conventional methods), 375 μL 0.08% (volume specific concentration) H 2 o 2 Solution, 325 μL inhibitor, 750 μL 0.05M guaiacol solution, 75 μL the above crude enzyme solution dilut...
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