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Clostridium AUH-JLC39 and application in conversion of tectorigenin

A technology of AUH-JLC39 and irisaxanthin, which is applied in Clostridium AUH-JLC39 and its application in the transformation of irisaxanthin, can solve the problems of no separation and artificial synthesis of dihydroirisaxanthin, and achieve new drug development The effect of promoting and solving microbial biosynthesis problems

Active Publication Date: 2014-04-16
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no report of isolating and artificially synthesizing dihydroirisaxanthin from nature, and there is no report of a single microbial strain that converts irisaxanthin into dihydroirisaxanthin

Method used

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  • Clostridium AUH-JLC39 and application in conversion of tectorigenin
  • Clostridium AUH-JLC39 and application in conversion of tectorigenin
  • Clostridium AUH-JLC39 and application in conversion of tectorigenin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] In this example, the crude irisaxanthin was used for conversion, and the crude irisaxanthin was obtained from the acid hydrolyzate of the methanol crude extract of kudzu flower through extraction with ethyl acetate.

[0054] 1. Isolation and cultivation of strain JLC39

[0055] (1) Collection and cultivation of chicken manure samples

[0056] Use a sterilized cotton swab to pick up fresh feces, put it into 1 ml of fresh BHI liquid medium, put it in an anaerobic workstation and cultivate it at 37°C for 24 hours, as a microbial flora for screening specific functional microbial strains;

[0057] (2) Isolation and cultivation of Clostridium AUH-JLC39

[0058] ①Single colony isolation culture

[0059] Use fresh BHI liquid medium to serially dilute the microbial flora that has been cultivated in the anaerobic workstation for 24 hours, and dilute to a concentration of 10 –1 , 10 –2 、10 –3 、10 –4 、10 –5 、10 –6 、10 –7 、10 –8 , and then 100 microliters of concentration ...

Embodiment 2

[0088]In this example, the crude irisaxanthin was used for transformation, which was obtained from the acid hydrolyzate of the crude methanol extract of Pueraria japonica through extraction with ethyl acetate; the isolation method of the strain JLC39 was the same as in Example 1.

[0089] (1) Cultivation of strain AUH-JLC39

[0090] Will –70 After freezing and thawing the low-temperature-preserved chicken intestinal isolate strain JLC39, inoculate 10% of the inoculum into a test tube containing fresh BHI liquid medium, and inoculate it in an anaerobic workstation for 37 After 20 hours of cultivation under high temperature, the bacterial solution in the test tube was flocculent and turbid. Then retransfer the turbid strain JLC39 in the test tube to a plastic centrifuge tube filled with fresh BHI liquid medium with 13% inoculum, and continue to cultivate in the anaerobic workstation for 20 hours as a seed solution;

[0091] (2) Co-cultivation of the substrate irisaxanthin wi...

Embodiment 3

[0094] In this example, pure irisaxanthin is used for transformation, which is obtained from acid hydrolyzate of kudzu flower methanol crude extract through high-efficiency liquid phase separation; the separation method of strain JLC39 is the same as that in Example 1.

[0095] (1) Cultivation of strain AUH-JLC39

[0096] Will –70 After freezing and thawing the low-temperature-preserved chicken intestinal isolate strain JLC39, it was inoculated into a test tube filled with fresh BHI liquid medium at a 13% inoculum size, and placed in an anaerobic workstation for 37 After 22 hours of cultivation under high temperature, the bacterial solution in the test tube was flocculent and turbid. Then retransfer the turbid strain JLC39 in the test tube to a plastic centrifuge tube filled with fresh BHI liquid medium with 12% inoculum, and continue to cultivate in the anaerobic workstation for 18 hours as a seed solution;

[0097] (2) Co-cultivation of the substrate irisaxanthin with st...

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Abstract

The invention discloses clostridium AUH-JLC39 with a preservation number of CGMCC No. 7221 and application of the clostridium AUH-JLC39 in conversion of tectorigenin. The application of the clostridium AUH-JLC39 in the conversion of the tectorigenin comprises the steps of grafting seed solution of bacterial strain JLC39 to a liquid medium according to 10-15% inoculation amount, adding substrate tectorigenin, standing and cultivating for 48-60 hours in an anaerobic working station, extracting culture twice using ethyl acetate with a same volume, evaporating the extracting liquid and separating and manufacturing the product on HPLC using a preparatory column. The bacterial strain can effectively convert the tectorigenin to dihydro tectorigenin, and the external clearing rate of the dihydro tectorigenin to DPPH is very remarkable and is higher than the substrate tectorigenin (p (0.01). The clostridium AUH-JLC39 solves the problem of microorganism synthesis of the dihydro tectorigenin and lack of resources, and has significant pushing effect of new medicine development.

Description

technical field [0001] The invention relates to a bacterial strain and its application, in particular to a Clostridium AUH-JLC39 and its application in irisaxanthin transformation. Background technique [0002] Pueraria lobata, also known as Pueraria japonica, is a leguminous plant of Pueraria mirifica ( Pueraria lobata (Wild.) Ohwi) or kudzu ( Pueraria thomsonii Benth. ) of dried flower buds. According to the records of the Pharmacopoeia of the People's Republic of China, kudzu flower is sweet and cool in nature, and it is used for relieving alcohol and refreshing the spleen, curing alcohol fever, polydipsia, and lack of appetite. It has been used for a long time to relieve symptoms such as vomiting after drinking. Pharmacological research results show that kudzu flower has various pharmacological activities such as protecting the liver, lowering blood sugar, lowering blood fat and estrogen. [0003] Pueraria japonica is a traditional Chinese medicine in my country. It...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/06C12R1/145
Inventor 王秀伶郭常亮邵子强刘明月
Owner HEBEI AGRICULTURAL UNIV.
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