Bacillus amyloliquefaciens and method for preparing succinyl ononin in nonaqueous phase

A technology of amylolytic spores and succinyl formonone, which is applied in the field of biocatalysis, can solve the problems of high cost and cumbersome steps, and achieve the effects of low cost, simple separation and improved solubility

Active Publication Date: 2016-11-30
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main method of preparing formononetin and its derivatives is chemical synthesis, which has high cost and cumbersome steps.

Method used

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  • Bacillus amyloliquefaciens and method for preparing succinyl ononin in nonaqueous phase
  • Bacillus amyloliquefaciens and method for preparing succinyl ononin in nonaqueous phase
  • Bacillus amyloliquefaciens and method for preparing succinyl ononin in nonaqueous phase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Screening and identification of strains for producing succinylated formononetin in non-aqueous phase

[0030] The soil samples for strain isolation were from the soils with severe chemical pollution and the soil of medicinal botanical gardens in various regions of Jiangsu Province. Extremophiles resistant to organic solvents were screened and isolated from the soil samples. Take about 1g of soil sample, put it into a conical flask filled with 20mL screening medium, 30°C, 180r min -1 Shake culture in a water bath for 24 hours, and transfer 3 times with 5% inoculum. After appropriate dilution of the culture solution, spread it with LB plate medium, incubate at 30°C for 24 hours, observe the colony shape, and then pick a single colony on the LB plate medium for streak isolation to obtain a single clone. numbered and stored in cryopreservation.

[0031] Using sucrose as the carbon source for the growth of organic solvent-resistant bacteria, adding 20% ​​(v / v) dimethylform...

Embodiment 2

[0037] Fermentation of Bacillus amyloliquefaciens FJ18 and preparation of resting cells

[0038] Fermentation of Bacillus amyloliquefaciens FJ18 was inoculated into the seed medium: yeast extract 5.0g / L, peptone 10.0g / L, NaCl 10.0g / L, pH7.0, cultured at 30°C, 200rpm for 12 hours. Expansion medium, fermentation medium, its components and contents are: sucrose 20g / L, yeast powder 15g / L.KH 2 PO 4 1.0g / L, CaCl 2 0.8g / L. The pH was adjusted to 8.0 with NaOH. The seed solution was inoculated into the expansion medium and the fermentation medium at 0.5% (v / v), and cultivated at 30° C. and 200 rpm for 12 hours. After centrifuging at 10000rpm for 15 minutes, the bacterial cells were collected, washed 1-2 times with physiological saline, and the resting cells of Bacillus amyloliquefaciens FJ18 were obtained.

Embodiment 3

[0040] The thalline cell fermentation broth in Example 2 was filtered to obtain wet thalline. The raw material solution, ie the reaction solution, is prepared with dimethyl sulfoxide, formononetin, sucrose and phosphate buffer. The ratio of organic solvent dimethyl sulfoxide in the reaction solution is 20% (v / v), formononetin 0.5g / L, the molar concentration of phosphate buffer is 150mmol / L, the pH of phosphate buffer is 8.0, and the sucrose concentration is 50g / L. Disperse the wet bacteria obtained above in the reaction solution, add it to the reactor, cultivate it at 30°C and 200 rpm for 24 hours, centrifuge at 10,000 rpm for 10 minutes to obtain the supernatant of the reaction solution, and detect and analyze the formononetin conversion by HPLC The rate is 97.2%.

[0041] The product is separated with a macroporous resin, and an appropriate amount of resin is taken, soaked in ethanol for 24 hours, and then resin fragments and sundries are removed. Wet Packing Rinse wit...

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Abstract

The invention relates to bacillus amyloliquefaciens and a method for preparing succinyl ononin in a nonaqueous phase, and belongs to the technical field of biological catalysis. The bacillus amyloliquefaciens is named bacillus amyloliquefaciens FJ18. The preservation unit is China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: M2016272. The invention also provides a method for preparing succinyl ononin in a nonaqueous phase by the strain. The strain can effectively catalyze fermlononetin to prepare succinyl ononin. Thus, the problem of scarcity of formononetin glucoside compounds is solved.

Description

technical field [0001] The invention belongs to the technical field of biocatalysis, and relates to a bacterial strain and a method for preparing succinyl formononetin, in particular to a bacillus amyloliquefaciens and a method for preparing succinyl formononetin in a non-aqueous phase. Background technique [0002] Flavonoids compounds (flavonoids compounds) are secondary metabolites of plants, which widely exist in natural plants in the form of free or combined with sugar as glycosides. Its pharmacological activity can prevent and treat cardiovascular and cerebrovascular diseases and respiratory system diseases, and has pharmacological effects such as anti-inflammatory, antibacterial, hypoglycemic, anti-oxidation, anti-radiation, anti-cancer, anti-tumor, and enhancing immunity. In recent years, the study of flavonoids has entered a new level. With the in-depth study of their structure-activity relationship, the mechanism of some pharmacological effects has been discovered,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/60C12R1/07
CPCC12P19/60C12N1/205C12R2001/07
Inventor 张森段金廒朱振华钱怡云
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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