Preparation method of genipin

A technology of genipin and preparation process, applied in microorganism-based methods, biochemical equipment and methods, immobilized on/in organic carriers, etc., can solve the problem that genipin is expensive and reduces the yield of genipin , the environment is easy to cause pollution and other problems, to achieve the effect of low cost, improve yield and overcome high cost

Inactive Publication Date: 2010-12-01
抚州市临川之信生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the preparation method of genipin generally adopts the purified β-glucosidase to convert geniposide such as: CN 101104745A; CN 101396455A; CN 101029066A, and then extract through ether and prepare by the method of acetone crystallization. The yield of genipin is low, the purity is not high, and the enzyme input cannot be recovered, so the cost is high, so the genipin sold on the market is expensive
In addition, the preparation of genipin needs to use a large amount of toxic organic solvents, which is easy to cause pollution to the environment; there are also reports on the use of microbial fermentation to convert geniposide to prepare genipin, such as Huo Lei, Su Jian, Shen Jing, etc., Natural Product Research and Development 2008, 20: 70-73; Wang Yonghong, Su Jian, Wang Jianfang, et a

Method used

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  • Preparation method of genipin
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  • Preparation method of genipin

Examples

Experimental program
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Effect test

Embodiment 1

[0036] (1) Ferment the enzyme-producing Aspergillus niger strain for 96 hours, take out the fermented liquid together with mycelium, and concentrate it with a hollow fiber ultrafilter with a molecular weight of 6000 to about one-third of the original volume, and add the final The gelatin with a concentration of 1% and the glutaraldehyde with a final concentration of 0.25% were cross-linked, the cross-linking time was 1 hour, and the cross-linking temperature was 10°C;

[0037] (2) Embedding the cross-linked enzyme and mycelia liquid with sodium alginate, the concentration of sodium alginate being 3%;

[0038] (3) Add the embedded mixed solution dropwise to the calcium chloride solution for fixation, the concentration of calcium chloride is 3%, and prepare immobilized enzyme pellets;

[0039] (4) Place the prepared immobilized enzyme pellets at room temperature in 0.5% calcium chloride solution for hardening, and the hardening time is 2 hours;

[0040] (5) Filter the hardened ...

Embodiment 2

[0044] (1) Ferment the enzyme-producing Aspergillus Usami strain for 120 hours, take out the fermented liquid together with mycelia, concentrate it with a hollow fiber ultrafilter with a molecular weight of 30,000, and concentrate it to about one-fifth of the original volume, and add the final Concentration is 0.5% chitosan and final concentration is 0.5% glutaraldehyde to carry out cross-linking, cross-linking time is 2 hours, and cross-linking temperature is 20 ℃;

[0045] (2) Embedding the cross-linked enzyme and mycelia liquid with sodium alginate, the concentration of sodium alginate being 3%;

[0046] (3) Add the embedded mixed solution dropwise to the calcium chloride solution for fixation, the concentration of calcium chloride is 3%, and prepare immobilized enzyme pellets;

[0047] (4) Place the prepared immobilized enzyme beads at room temperature in 0.5% calcium chloride solution for hardening, and the hardening time is 3 hours;

[0048] (5) Filter the hardened immo...

Embodiment 3

[0052] (1) Ferment the enzyme-producing Rhizopus oryzae strain for 96 hours, take out the fermented liquid together with mycelia, add a final concentration of 2% gelatin and a final concentration of 1% glutaraldehyde for cross-linking, and the cross-linking time is 1 hour , the crosslinking temperature is 10°C;

[0053] (2) Embedding the cross-linked enzyme and mycelia liquid with sodium alginate, the concentration of sodium alginate being 3%;

[0054] (3) Add the embedded mixed solution dropwise to the calcium chloride solution for fixation, the concentration of calcium chloride is 4%, and prepare immobilized enzyme pellets;

[0055] (4) Place the prepared immobilized enzyme pellets at room temperature in 0.5% calcium chloride solution for hardening, and the hardening time is 2 hours;

[0056] (5) Filter the hardened immobilized enzyme and store it at 4°C for later use;

[0057] (6) Wet-pack the immobilized enzyme into a column, then prepare 2% geniposide solution, adjust t...

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Abstract

The invention belongs to the biotechnology field and relates to a preparation method of genipin. The method comprises a preparation process of immobilized enzyme, a transformation process of gardenoside and a separation and purification process of genipin and is characterized by fermenting the strains for producing beta-glucosidase, collecting the fermentation liquor after reaching the enzyme producing peak, co-immobilizing enzyme and cells or enzyme and hyphae by combining embedding with crosslinking to prepare the immobilized enzyme, utilizing a packed-bed or stirred reactor to carry out catalyzed hydrolysis on the gardenoside and obtaining genipin after purification. The invention dispenses with separation and purification of the fermentation liquor, the immobilized enzyme obtained by preparation has high activity still keeping over 50% after operation for 960h and produces less pollution to the environment, genipin has high yield and the method has obvious advantages compared with other methods.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a preparation method of genipin, in particular to a method for preparing genipin by co-immobilizing enzyme and cell biocatalyzed hydrolysis of geniposide. Background technique [0002] Genipin is an iridoid compound obtained by hydrolyzing geniposide, also known as geniposide, with β-glucosidase. Genipin has multiple uses. One is that it can react with amino acids to produce blue pigment. This natural blue pigment can be used as a food additive. It is non-toxic and stable, and has been widely used abroad; It can be used as a natural biological cross-linking agent. In modern biomedical practice, in order to reduce the rejection of organisms to implanted xenogeneic biological tissues or artificial substitutes, and to increase the biocompatibility and mechanical properties of biomaterials, cross-linked To achieve the goal, most of the traditional cross-linking agents are chemically synth...

Claims

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Application Information

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IPC IPC(8): C12P17/06C12N11/04C12N11/10C12N11/02C12R1/685C12R1/845C12R1/66
Inventor 梁华正唐伯辰贺玉兰陈贺徐尤智谢键泓曾彦雯
Owner 抚州市临川之信生物科技有限公司
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