Method for preparing dihydro-dehydro-di(coniferyl alcohol)glucoside (HMZG) through biotransformation

A technology for coniferyl glycoside and biotransformation, applied in the biological field, can solve the problems of high cost, difficult to prepare in large quantities, pollute the environment, etc., and achieve the effects of low cost and easy operation

Active Publication Date: 2016-08-17
上海椿龄健康管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because its content in plants is very small, it is difficult to extract and separate from plants to prepare in large quantities; using chemical methods to glycosylate dihydro-dehydrobiconiferyl

Method used

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  • Method for preparing dihydro-dehydro-di(coniferyl alcohol)glucoside (HMZG) through biotransformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Preparation of HMZG-1, HMZG-2 and HMZG-3 (cultured with PDA medium)

[0021] The slant (i.e. bacterial classification) was inoculated in the shake flask seed medium, cultivated for 24 hours at 26°C, and the shake flask rotating speed was 180r / min; access the PDA transformation medium (potato 200g / L, KH 2 PO 4 3g / L, MgSO 4 0.73g / L, VB 1 10mg / L, sucrose 25g / L, pH 7.0), while adding 0.2mg / mL of substrate, at 26°C, the rotation speed was 180r / min for 72h. The conversion rate of the three conversion products HMZG-1, HMZG-2 and HMZG-3 was detected by HPLC. The detection condition was acetonitrile-water gradient elution, and the elution program was: 0-10min 15%-24% acetonitrile, 10-25min 24%-30% acetonitrile, 30% acetonitrile for 25-30min; flow rate 1.0mL / min; column temperature 25°C; detection wavelength 203nm; injection volume 10μL. The retention times of HMZG-1, HMZG-2 and HMZG-3 were 12.306min, 14.817min and 15.359min respectively, and the conversion rates were 17.3...

Embodiment 2

[0023] Preparation of HMZG-1, HMZG-2 and HMZG-3 (cultured with BPY medium)

[0024] The slant (i.e. bacterial classification) was inoculated in the shake flask seed medium, cultivated at 26°C for 24 hours, and the shake flask rotating speed was 180r / min; insert BPY transformation medium (beef extract 5.0g / L) by 2% inoculum , peptone 10.0g / L, yeast extract 5.0g / L, NaCl 5.0g / L, glucose 10.0g / L, pH 7.0), add 0.2mg / mL substrate at the same time, at 26°C, the speed is 180r / min Fermentation and culture under the conditions for 72h. The conversion rates of the three conversion products HMZG-1, HMZG-2 and HMZG-3 were detected by HPLC. The detection conditions were the same as in Example 1, and the conversion rates were 13.20%, 10.36%, and 50.24%, respectively.

Embodiment 3

[0026] Preparation of HMZG-1, HMZG-2 and HMZG-3 (cultured with NA medium)

[0027] The slant (i.e. bacterial classification) was inoculated in the shake flask seed medium, cultivated at 26°C for 24 hours, and the shake flask rotating speed was 180r / min; insert NA transformation medium (beef extract 3.0g / L) by 2% inoculum , peptone 10.0g / L, NaCl 5.0g / L, agar 18.0g / L, pH 7.0), and 0.2mg / mL substrate was added at the same time, and the fermentation was carried out at 26°C and the rotation speed was 180r / min for 72h. The conversion rates of the three conversion products HMZG-1, HMZG-2 and HMZG-3 were detected by HPLC. The detection conditions were the same as in Example 1, and the conversion rates were 10.50%, 13.51%, and 45.98%, respectively.

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Abstract

The invention belongs to the biotechnology field, and particularly relates to a method for preparing dihydro-dehydro-di(coniferyl alcohol)glucoside (HMZG) through biotransformation. The dihydro-dehydro-di(coniferyl alcohol)glucoside (HMZG) is obtained from a strain with the preservation number of Bacillus subtilis ATCC 6633 through biotransformation, and three glucoside pure products (HMZG-1, HMZG-2 and HMZG-3) are obtained through repeated silicagel column chromatography purification. Therefore, the biotransformation method for efficiently obtaining the dihydro-dehydro-di(coniferyl alcohol)glucoside (HMZG) is established.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing dihydrodehydrobisconiferyl glucomonoside (HMZG) through biotransformation. Background technique [0002] Dihydrodehydrobiconiferyl glucomonoside (HMZG) is a trace active ingredient in medicinal plants such as Acanthopanax, Polygonum cuspidatum, Elderberry, Herba Cortex, etc. It has a wide range of pharmacological activities, such as liver protection, anti-oxidation, anti-inflammation , anti-fungal, anti-HSV and anti-virus, etc. Belongs to the benzodihydrofuran class neolignan glucose monoglycoside. Because its content in plants is very small, it is difficult to extract and separate from plants to prepare in large quantities; using chemical methods to glycosylate dihydro-dehydrobiconiferyl alcohol to obtain its glucose monoglycoside has complicated steps, high yield and low cost, and may pollute the environment. The development and utilization of s...

Claims

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Application Information

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IPC IPC(8): C12P19/44C12R1/125
CPCC12P19/44
Inventor 刘吉华余伯阳盖晓红
Owner 上海椿龄健康管理有限公司
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