AUH-JLC159 and transforming method thereof for preparation of (-)-5-OH-equol

A technology of AUH-JLC159, -5-OH-, applied in the field of Snaekia and its preparation of -5-OH-equol in genistein conversion

Active Publication Date: 2014-08-20
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Similar to equol, 5-OH-EQ is also a chiral compound, and there should be two enantiomers R- and S- in theory, although Japanese and German scholars have successively isolated 5-OH-EQ producing bacteria from human and mouse feces, but so far it is not clear whether the 5-OH-EQ produced by the bacterial transformation of genistein is also a single enantiomers (i.e. only the S-form or only the R-form); in addition, it is still unclear whether the 5-OH-EQ produced by bacterial transformation There is optical activity, if there is optical activity, it is not clear whether it is left-handed or right-handed

Method used

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  • AUH-JLC159 and transforming method thereof for preparation of (-)-5-OH-equol
  • AUH-JLC159 and transforming method thereof for preparation of (-)-5-OH-equol
  • AUH-JLC159 and transforming method thereof for preparation of (-)-5-OH-equol

Examples

Experimental program
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Effect test

Embodiment 1

[0045] 1. Isolation and cultivation of strain JLC159

[0046] (1) Collection and cultivation of rooster manure samples

[0047] Use a sterilized cotton swab to pick a fresh fecal sample, put it into 1 ml of fresh BHI liquid medium, and place it in an anaerobic workstation at 37°C, as a microbial flora for screening specific functional microbial strains;

[0048] (2) Isolation and cultivation of transformed strains

[0049] ①Single colony isolation culture

[0050] Use the microbial flora in the fresh BHI liquid medium for gradient dilution, 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 were respectively 10 –5 、10 –6 、10 –7 、10 –8 The dilution of microbial flora was uniformly coated on the pre-prepared BHI solid medium, and the BHI solid medium coated with the dilution of microbial flora was placed in an anaerobic workstation for 48 hours. Pick dozens to hundreds of single colon...

Embodiment 2

[0084] The isolation method of bacterial strain JLC159 is the same as that in Example 1.

[0085] In the anaerobic workstation, the seed liquid of the above-mentioned pre-cultivated strain JLC159 was transferred to a 250-ml Erlenmeyer flask containing 100 ml of liquid medium at a 15% inoculum size, and 40 mmol / L of genistein was added at the same time. 1.25 ml of mother liquor was cultured in an anaerobic workstation for 3 days. Use an equal volume of ethyl acetate to extract the culture in the Erlenmeyer flask twice, filter the extract and evaporate to dryness on a rotary evaporator, then add 100% methanol solution, pass through an organic membrane with a pore size of 0.45 microns, and use a preparative column to perform HPLC analysis. Separation and preparation were carried out above to obtain a conversion rate of (-)-5-OH-EQ of 83.3%.

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Abstract

The invention discloses a Shrekia AUH-JLC159 with a preservation number of CGMCC No. 5304 and an application thereof in preparing (-)-5-OH-equol. (1) Cultivation of strain AUH-JLC159; (2) Co-cultivation of genistein with strain AUH-JLC159 and separation and purification of metabolites: the seed liquid of strain AUH-JLC159 was transferred to BHI medium and worked in anaerobic The substrate genistein can be converted into (-)-5-OH-equol by culturing in the station for 2 to 3 days; the metabolites are separated and purified. The Shrekella AUH-JLC159 of the present invention can efficiently convert the substrate genistein into 5-OH-EQ under anaerobic conditions; it solves the problem of microbial biosynthesis and resource scarcity of 5-OH-EQ, and is effective for 5 -OH-EQ physiological, pharmacological activity and new drug research and development have a great role in promoting.

Description

technical field [0001] The invention relates to a bacterium and an application method thereof, in particular to a snagella and an application method thereof in preparing (-)-5-OH-equol by converting genistein. Background technique [0002] Most of the isoflavones in natural soybeans exist in the form of bound glycosides, and there are only three kinds of soybean isoflavones in the form of free aglycones, namely daidzein, genistein and genistein (Glycitein), of which daidzein and genistein account for 95% to 97% of the total soybean isoflavone aglycones, and are the main components of soybean isoflavone aglycones. A large number of epidemiological and laboratory studies have shown that soybean isoflavones have obvious antioxidant, anti-inflammatory, anti-tumor effects, relieve menopausal symptoms, prevent and treat osteoporosis, and reduce the incidence of cardiovascular and cerebrovascular diseases. The physiological function of the human body has aroused widespread concer...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/06C12R1/01
Inventor 王秀伶刘子光邵子强梁晓琳张红蕾郭常亮
Owner HEBEI AGRICULTURAL UNIV.
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