A kind of Raoultella pan22x and its application
A CGMCCNO.12208, seed technology, applied in the field of anti-bacterial quorum sensing extracts, to achieve the effects of obvious inhibition of quorum sensing, simple and easy-to-control preparation process, and simple culture conditions
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experiment example 1
[0025] Experimental example 1: Isolation, purification and identification of Raoultella sp. pan22x
[0026] (1) Isolation and cultivation of marine bacteria and fungi
[0027] 1) Isolation and culture of bacteria
[0028] Sample collection: In April 2014, seaweed samples were collected from the East China Sea at 29.9956 degrees north latitude and 122.3942 degrees east longitude.
[0029] Treatment of seaweed samples: Take 2g of seaweed, rinse it with sterile water for 3 times, put it into a sterile mortar and grind it, add 2ml of sterile water and mix it as the original solution, and then dilute it by 7 times to get 1×10 -1 ~1×10 -7 A total of 7 samples were spread on the bacterial isolation plate, cultured at 37°C for 20 hours, and single colonies pan22x, P-16, pan26, pan17, 10 -5 New, observe deweighting.
[0030] Composition of bacteria separation plate per liter: yeast extract 5g, peptone 10g, NaCl 5g, agar 15-20g, artificial seawater 1L, pH7.1-7.3, 1×10 5 Pa steriliz...
Embodiment 2
[0051] Embodiment 2. The fermentation culture of Raoultella pan22x and the preparation of antibacterial agent
[0052] (1) Incline cultivation
[0053] Inoculate Raoultella pan22x into the slant medium and culture it at 30-37°C for 18-20h to obtain the slant bacteria; the final concentration of each liter of the slant medium is composed of: yeast extract 5g, peptone 10g, NaCl 5g, agar 15 ~20g, artificial seawater 1L (composition is the same as Example 1), pH7.1~7.3, 1×10 5 Pa sterilize for 20min and set aside;
[0054] (2) Seed cultivation
[0055] Inoculate the slant bacteria into the seed medium, culture at 30-37°C for 18-20h, and obtain the seed liquid; the final concentration of the seed medium per liter is composed of: 5g of yeast extract, 10g of peptone, 5g of NaCl, and 1L of artificial seawater (composition is the same as embodiment 1), pH7.1~7.3, 1 * 10 5 Pa sterilize for 20min and set aside;
[0056] (3) Fermentation culture
[0057] Inoculate the seed liquid in...
experiment example 3
[0059] Experimental example 3 Quorum sensing inhibition of C.violaceum ATCC 12472 by Raoultella pan22x fermented in different media
[0060] (1) Incline cultivation
[0061] Inoculate Raoultella pan22x into the slant medium and culture it at 30-37°C for 18-20h to obtain the slant bacteria; use five kinds of medium for cultivation, respectively: LB fresh water: 5g of yeast extract, 10g of peptone, NaCl 5g, 1L deionized water, 15g agar powder; pH7.1~7.3, 1×10 5 Pa sterilize for 20min for later use.
[0062] Nutrient agar seawater: peptone 10g, beef extract 5g, sodium chloride 5g, artificial seawater 1L (composition is the same as in Example 1), agar powder 15g; pH7.1~7.3, 1×10 5 Pa sterilize for 20min for later use.
[0063] Nutrient agar fresh water: peptone 10g, beef extract 5g, sodium chloride 5g, deionized water 1L, pH7.1~7.3, 1×10 5 Pa sterilize for 20min for later use.
[0064] NB seawater: peptone 5g, NaCl 5g, yeast extract 1.5g, beef extract 1.5g, agar powder 15g; p...
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