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Method for efficiently transforming immobilized streptomyces griseus cells to prepare corydalmine

A technology of Streptomyces griseus and immobilized cells, which is applied in the biological field, can solve the problems of limiting the input amount of the transformation substrate tetrahydropalmatine, and the transformation and fermentation period is long, and achieves low production cost, low cost and simple process operation. Effect

Inactive Publication Date: 2014-07-30
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Above method utilizes unimmobilized cell to carry out the conversion of alkaloid tetrahydropalmatine, because alkaloid tetrahydropalmatine has certain toxic effect to the growth of Streptomyces griseus cell, has limited the transformation substrate tetrahydropalmatine Feeding amount, and the conversion fermentation cycle is longer as 96h

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The preparation of embodiment 1 Streptomyces griseus immobilized cells

[0019] (1) Cultivation of Streptomyces griseus (Streptomyces griseus) ATCC-13273 seed solution

[0020] Streptomyces griseus fermentation medium was used to inoculate Streptomyces griseus (Streptomyces griseus) ATCC-13273 from slant culture after sterilization, and cultured on a shaker at 28°C and 180rpm for 24 hours as a seed liquid for later use; among them, Streptomyces griseus fermented The medium formula is: 3% corn flour, 0.76% soybean meal, 0.5% yeast powder, 0.5% K 2 HPO 4 , 0.12% Glucose, 0.5% NaCl, 0.01% MgSO 4 7H 2 O, the initial pH is 6.3;

[0021] (2) Pretreatment of polyurethane foam carrier

[0022] Cut the polyurethane foam into a cube with a side length of 5 mm, soak it in 5% hydrochloric acid solution for 24 hours, wash it repeatedly with distilled water until neutral, then soak it with 5% sodium hydroxide solution for 24 hours, wash it repeatedly with distilled water until n...

Embodiment 2

[0025] Example 2 Using Immobilized Streptomyces griseus Cells to Transform Tetrahydropalmatine in a Shake Flask to Prepare Corydalamine

[0026] Take a 250mL Erlenmeyer flask and add 40mL of immobilized Streptomyces griseus cell transformation medium to sterilize, then add 0.2g / L of immobilized cells, 0.8g / L of transformation substrate tetrahydropalmatine, and transform in a shaker at 160rpm and 28°C After 24 hours, the transformed fermentation broth was used for the detection of the conversion yield of Corydalis damming, and the immobilized cells were taken out and used for transformation and preparation of Corydalis damming under the same conditions as above. The immobilized cells were reused 15 times, and the high performance liquid chromatography showed that Immobilized Streptomyces griseus is used for shake flask transformation, and the conversion yield of Corydalis dammine is greater than 59% every time in the repeated use process; among them, the transformation medium of...

Embodiment 3

[0027] Example 3 Using immobilized Streptomyces griseus cells to transform tetrahydropalmatine in a fermenter to prepare Corydalamine

[0028] Add 60L Streptomyces griseus immobilized cell transformation medium to a 100L fermenter, add 0.3g / L immobilized Streptomyces griseus cells after sterilization, 0.8g / L transformation substrate tetrahydropalmatine, 28°C, stirring speed 150rpm , ventilation rate 1.2vvm transformation and fermentation for 36h, release the fermented liquid for the separation and preparation of Corydalis damming, add the sterilized transformation medium and transformation substrate in the fermenter and transform and prepare Corydalis damming under the same conditions as above, fix The immobilized cells were reused 16 times, and the high-performance liquid chromatography detection showed that the conversion yield of Corydalis damming was greater than 56% every time when the immobilized Streptomyces griseus was used in the transformation process of the fermenter...

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Abstract

The invention discloses a method for efficiently transforming immobilized streptomyces griseus cells to prepare corydalmine. The method comprises the following steps: culturing streptomyces griseus with a collection number of ATCC-13273 by a fermentation culture medium, pre-treating a polyurethane foam carrier, culturing and immobilizing cells and efficiently transforming immobilized streptomyces griseus ATCC-13273 cells to prepare the corydalmine. The method disclosed by the invention solves the problems that the corydalmine prepared by an existing non-immobilized cell transformation method is long in transformation period, low in transformation substrate feeding concentration and low in corydalmine transformation yield. And moreover, the cell immobilizing method is simple, and low in cost; the immobilized cells can be repeatedly used to realize continuous transformation preparation of the corydalmine, and therefore, the method has good economic benefits.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for efficiently transforming Streptomyces griseus (Streptomyces griseus) ATCC-13273 cells into immobilized polyurethane foam to prepare Corydalis damming. Background technique [0002] Corydalis damamine is a natural trace active ingredient with good analgesic activity, and has the characteristics of high-efficiency and rapid analgesia, but its natural sources are very limited, which limits its application in the field of medicine. We have previously reported the method of biotransformation preparation of Levocordydamine (Pharmaceutical Biotechnology, 2003, 10 (3): 165-168), and applied for the biotransformation preparation method of Corydalamine and its application in the preparation of analgesics The application in (Chinese patent ZL02138363.4) and a biotransformation fermentation process for efficient production of Corydalis damming (Chinese patent ZL201110125328.2) paten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/545
Inventor 刘吉华索成林余伯阳
Owner CHINA PHARM UNIV
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