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Raoultella sp. pan22x and application thereof

A technology of CGMCCNO.12208 and seeds is applied in the field of anti-bacterial quorum sensing extracts, and achieves the effects of simple culture conditions, high yield, and obvious inhibition of quorum sensing.

Active Publication Date: 2016-08-17
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the widespread use of antibiotics, the emergence of bacterial resistance has become a new bottleneck in finding a new broad-spectrum antibacterial

Method used

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  • Raoultella sp. pan22x and application thereof
  • Raoultella sp. pan22x and application thereof
  • Raoultella sp. pan22x and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The present invention relates to a Raoultella sp. pan22x and application thereof to production of a bacteriostatic agent for inhibiting purple pigment generation. The marine Raoultella sp. pan22x is preserved in China General Microbiological Culture Collection Center, Institute of Microbiology, Chinese Academy of Sciences, No 3, Yard No.1, Beichen West Road, Chaoyang District, Beijing (zip code 100101) on the date of March 11, 2016, and has an accession number CGMCC NO. 12208. The bacteriostatic agent of the invention can effectively inhibit purple pigment production of C.violaceum ATCC12472 and C. violaceum CV026 in the sample addition amount of 0-2.0mg. The antibacterial agent has quorum sensing inhibition effect showing concentration dependence; along with the increase of sample addition amount, the inhibition effect is enhanced; in concentration range of 0.1-0.3mg / mL, the inhibition rate of purple pigment production of C. violaceum ATCC 12472 reaches 54% without affecting cell growth. The Raoultella sp. pan22x fermentation productscan be used to research on the prevention of purple bacterium infections, and the development of new antimicrobial agents in the medical field.

Description

(1) Technical field [0001] The invention belongs to the field of marine microorganisms, in particular to an antibacterial quorum sensing extract prepared from Raoultella sp. pan22x and its application. (2) Background technology [0002] Bacterial infection has always been a threat to human health, and a complete eradication of bacterial infection is a major problem plaguing human beings. Since the German scientist Robert Koch confirmed the theory of the bacterial origin of infectious diseases in the late 19th century, people have gained a new understanding of the treatment of infectious diseases, and the search for drugs for bacterial infections has a relatively clear goal, and the discovery of antibiotics represented by penicillin Bacterial infection was once effectively restrained. However, with the widespread use of antibiotics, the emergence of bacterial resistance has become a new bottleneck in finding a new broad-spectrum antibacterial. The process of discovering new...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/04A61K35/74A61P31/04C12R1/01
CPCA61K35/74C12N1/20C12P1/04C12N1/205C12R2001/01
Inventor 王鸿平夏婷陈苏潘秋
Owner ZHEJIANG UNIV OF TECH
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