Method for preparing coronatine and special strain thereof

A technology of coronatine and variants, applied in the field of coronatine preparation, can solve the problems of lack of excellent and high-yielding strains and high cost of coronatine fermentation production, and achieve the effect of increasing yield

Active Publication Date: 2009-01-07
CHENGDU NEWSUN CROPSCI
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Problems solved by technology

The main reason for restricting its industrial application is that the current fermentation p...

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  • Method for preparing coronatine and special strain thereof
  • Method for preparing coronatine and special strain thereof
  • Method for preparing coronatine and special strain thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the construction of Pseudomonas syringae pathogenic variant soybean recombinant bacteria

[0054] 1. Construction of Escherichia coli containing plasmid vector pRL1063a

[0055]The plasmid vector pRL1063a (Cloning and functional analysis ofcysI gene involved in siderophores biosyntheses in Pseudomonas mossel ii E1, Huang WH, Ding YQ, Yao LT, Du ZB, Du BH, Department of Microbiology, College of life sciences, Shandong Agricultural University, Tai'an 271018, China.hwh7252@163.com) (China Agricultural University) was introduced into Escherichia coli S17-1, and then screened with streptomycin to obtain positive recombinant Escherichia coli containing the plasmid vector pRL1063a. It was named Escherichia coli S17-1 / pRL1063a.

[0056] The structure of plasmid pRL1063a is as follows Figure 4 shown.

[0057] 2. Preparation of recombinant bacteria

[0058] Pseudomonas syringae pv.glycinea (Carol Bender, David Palmer, Alejandro - Vázquez, Vi dhya Rangaswamy, ...

Embodiment 2

[0066] Embodiment 2, fermentation prepares coronatin

[0067] 1. Fermentation in a triangular flask

[0068] 1. Fermentation

[0069] Inoculate Pseudomonas syringae pathogenic strain of soybean MW123 (preservation number CGMCC No.2533) into MG liquid medium containing kanamycin at a final concentration of 10 μg / L and streptomycin at a final concentration of 30 μg / L , cultivated on a shaker at a temperature of 26°C and a rotation speed of 260rpm, until the OD of the culture medium 600 The value reached 0.7 and a seed solution was obtained.

[0070] According to the inoculum amount of 2.5% (volume ratio), above-mentioned seed solution is inoculated in the 500ml Erlenmeyer flask that 200ml fermentation medium is housed, is 21 ℃ at temperature, and the shaking table that rotating speed is 260rpm cultivates 5 days;

[0071] The fermentation medium was HSC medium containing kanamycin at a final concentration of 10 μg / L and streptomycin at a final concentration of 30 μg / L.

[007...

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Abstract

The invention discloses a method for preparing coronatine and a special bacterium thereof. The bacterium provided by the invention is a pseudomonas syringae pv.glycinea with a preservation number of CGMCC No.2533. The bacterium MW123 (with the preservation number of CGMCC No.2533) is an excellent bacterium of a coronatine with high output. Test result shows that the method for using the bacterium MW123 (with the preservation number of CGMCC No.2533) to prepare the coronatine has the advantage of high and stable output; the output can achieve 180mg/L to 200mg/L offermentation broth. Compared with the original bateriusm, the output is improved by 3 to 4 times. Therefore, the method of the invention for preparing the coronatine solves the difficulty of low output of the coronatine of the existing fermentation production and is applicable for industrial popularization and application.

Description

technical field [0001] The invention relates to a method for preparing coronatin and a special bacterial strain thereof. Background technique [0002] Coronatine is composed of an amino acid-containing coronamic acid and a polyketide-structured coronafacic acid linked by an amide bond, and its molecular formula is C 18 h 25 NO 4 , with a molecular weight of 319. Studies have shown that coronatin has similar functions to the plant growth regulator substances abscisic acid and jasmonic acid, and its activity is thousands of times that of the latter two. substitution. Coronatine is a pure natural product, and its application in production will not cause pollution to the environment. It is an environmentally friendly biological regulator. [0003] So far, there is no batch product of coronatin in the world, and only a small amount of samples are purified by individual laboratories for research needs. The main reason for limiting its industrial application is that the curre...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/02C12R1/38
Inventor 李召虎吴慧玲章家长段留生
Owner CHENGDU NEWSUN CROPSCI
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