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Method for producing paeonin metabolite-I by short lactobacillin fermentation

A technology of paeoniflorin metabolites and Lactobacillus brevis, which is applied in the field of biochemical engineering, can solve the problems of unsuitability for industrialization, no report of yield rate, long conversion time, etc., and achieve easy control of the process, mature separation method, and fast production cycle Effect

Inactive Publication Date: 2010-01-27
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In 1985, Tsuneo Namba of Japan reported the preparation process of paeoniflorin metabolite-I obtained by fermenting paeoniflorin in vitro with intestinal bacteria. And there is no yield report, not suitable for industrialization

Method used

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  • Method for producing paeonin metabolite-I by short lactobacillin fermentation
  • Method for producing paeonin metabolite-I by short lactobacillin fermentation

Examples

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Effect test

Embodiment 1

[0026] The components and contents of seed medium, expansion medium and fermentation medium are: tomato juice 200g / L, peptone 7.5g / L, yeast extract 7.5g / L, glucose 10g / L, Tween 80 0.3ml / L L, adjust the pH value to 7.0-8.5 with 10% NaOH, and sterilize at 115°C for 30 minutes.

[0027] Take out 10ml of Lactobacillus brevis AS 1.12 from the anaerobic tube, inoculate it into 100ml of seed medium, and culture it anaerobically at 37°C for 24 hours, and culture 5 copies in total. Inoculate 5 parts of the culture solution into 5L expansion medium, expand the culture for 24 hours, inoculate the culture solution into 45L fermentation medium, culture anaerobically at 37°C for 24 hours, collect the cells by centrifugation, wash once with normal saline, and inoculate Suspend in 0.05 mol / L phosphate buffer (concentration: 40 g wet cells / L) and add paeoniflorin to make the concentration 1 mg / ml, transform at 37° C. for 6 hours. The conversion solution was extracted 4 times with ethyl acetat...

Embodiment 2

[0029] The components and contents of seed culture medium, expansion medium and fermentation medium are the same as in Example 1.

[0030] Take out 10ml of Lactobacillus brevis AS 1.12 from the anaerobic tube, inoculate it into 100ml of seed medium, and culture it anaerobically at 37°C for 24 hours. Inoculate the culture solution in 1L expansion medium, expand the culture for 24 hours, inoculate the culture solution in 10L fermentation medium, culture anaerobically at 37°C for 24 hours, collect the cells by centrifugation, wash once with normal saline, and suspend in Add benzoyl paeoniflorin to 0.05 mol / L phosphate buffer solution (concentration: 40 g wet cells / L) to make the concentration 1 mg / ml, and transform at 37° C. for 6 hours. The conversion solution was extracted 4 times with ethyl acetate, and the extracts were combined and concentrated to dryness to obtain 4.5 g of extract. The extract was subjected to ordinary silica gel column chromatography to obtain 0.1 g of pa...

Embodiment 3

[0032] The components and contents of seed culture medium, expansion medium and fermentation medium are the same as in Example 1.

[0033] Take out 10ml of Lactobacillus brevis AS 1.12 from the anaerobic tube, inoculate it into 100ml of seed medium, and culture it anaerobically at 37°C for 24 hours. Inoculate the culture solution in 1L expansion medium, expand the culture for 24 hours, inoculate the culture solution in 10L fermentation medium, culture anaerobically at 37°C for 24 hours, collect the cells by centrifugation, wash once with normal saline, and suspend in Add oxidized paeoniflorin to 0.05 mol / L phosphate buffer solution (concentration: 40 g wet cells / L) to make the concentration 1 mg / ml, and transform at 37° C. for 6 hours. The conversion solution was extracted 4 times with ethyl acetate, and the extracts were combined and concentrated to dryness to obtain 3.2 g of extract. The extract was subjected to ordinary silica gel column chromatography to obtain 0.07 g of ...

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Abstract

A method for producing paeonoside metabolic hormone-I by fermenting short Lactobacillus is carried out by inducing L.brevis AS 1.12 into culture medium with tomato juice 200g / L, protein peptone 7.5g / L, yeast paste 7.5g / L, glucose 10g / L, Tween-80 0.3 ml / L and pH=7.0-8.5, anaerobic culturing at 25-37 degree for 12-24 hrs, centrifugal collecting bacterial cells, washing by normal saline, centrifuging, dispersing obtained cells into buffer liquid of 0.05M phosphate, adding into substrate paeonoside or its analogs hydroxy-paeonoside or methyl-paeonoside or benzoyl-paeonoside or mixture of paeonoside and its analogs, converting at 30-40 degree for 2-24 hrs, extracting by ethyl acetate, drying concentrated extract to obtain concrete, chromatographying by silica-gel column to obtain final product. It's simple, cheap, efficient, fast and controllable and has friendly environment. It can be used for anti-inflammatory, stress ulcer, analgesic and antispasmodic, ecstatic coronary blood vessel, myocardial ischemia and platelet aggregation-inhibition.

Description

technical field [0001] The invention belongs to the field of biochemical engineering, and relates to a method for producing paeoniflorin metabolite-I by fermenting lactobacillus. Background technique [0002] Paeoniflorin is the main active ingredient of Paeoniflorin, which has various effects such as analgesic and antispasmodic, anti-inflammatory, anti-stress ulcer, expansion of coronary vessels, anti-acute myocardial ischemia and inhibition of platelet aggregation. Recent studies have shown that after oral administration of paeoniflorin, there is little direct absorption and low bioavailability, suggesting that paeoniflorin is first degraded by intestinal bacteria in the intestinal tract, and its metabolites play the pharmacological role of paeoniflorin. Japan's Masao Hattori et al. first reported that paeoniflorin was converted into paeoniflorin metabolin-I, II, III (see Figure 1 for Paeonimatabolin-I, II, III) in human intestinal bacteria in vitro culture medium, paeonif...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/04A61K31/357A61P9/10A61P7/02A61P29/00C12R1/24
Inventor 王晓玲官家发彭树林范成英焦威丁立生
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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