Preparation method of genipin

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...

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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 al. Enzyme and MicrobialTechnology, 2008, 42: 440-444, but there is a relatively big defect in this method is that there are more amino acids and protein substances in the fermentation liquid during microbial fermentation. When geniposide is hydrolyzed to generate genipin, Genipin will react with amino acids to generate blue pigment, which greatly reduces the yield of genipin; foreign countries have reported the technology of using immobilized β-glucosidase to convert geniposide to prepare genipin, such as ShigeakiFUJIKAWA, Tomoko YOKOTA, Kunimasa KOGA, et al.Biotechnology Letters, 1987,9 (10): 697-702, conversion rate can reach more than 95%, and the half-life of immobilized enzyme reaches 600 hours, but this method needs to obtain a large amount of pure β-glucoside Enzymes and therefore more expensive

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