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1. WO2020107629 - USE OF HIGH-SPEED COUNTERCURRENT CHROMATOGRAPH IN BIOCONVERSION OF GLYCOSIDE COMPONENT

Publication Number WO/2020/107629
Publication Date 04.06.2020
International Application No. PCT/CN2018/125458
International Filing Date 29.12.2018
IPC
C12P 17/06 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
17Preparation of heterocyclic carbon compounds with only O, N, S, Se, or Te as ring hetero atoms
02Oxygen as only ring hetero atoms
06containing a six-membered hetero ring, e.g. fluorescein
CPC
C12M 21/18
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
21Bioreactors or fermenters specially adapted for specific uses
18Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
C12M 27/02
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
27Means for mixing, agitating or circulating fluids in the vessel
02Stirrer or mobile mixing elements
C12P 17/06
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
17Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
02Oxygen as only ring hetero atoms
06containing a six-membered hetero ring, e.g. fluorescein
Applicants
  • 山东省分析测试中心 SHANDONG ANALYSIS AND TEST CENTER [CN]/[CN]
Inventors
  • 王岱杰 WANG, Daijie
  • 王晓 WANG, Xiao
  • 耿岩玲 GENG, Yanling
  • 赵恒强 ZHAO, Hengqiang
  • 崔莉 CUI, Li
  • 闫慧娇 YAN, Huijiao
Agents
  • 济南圣达知识产权代理有限公司 JINAN SHENGDA INTELLECTUAL PROPERTY AGENCY CO., LTD
Priority Data
201811435142.528.11.2018CN
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) USE OF HIGH-SPEED COUNTERCURRENT CHROMATOGRAPH IN BIOCONVERSION OF GLYCOSIDE COMPONENT
(FR) UTILISATION D'UN CHROMATOGRAPHE À CONTRE-COURANT À GRANDE VITESSE DANS LA BIOCONVERSION D'UN COMPOSANT GLYCOSIDE
(ZH) 高速逆流色谱仪在糖苷类成分生物转化中的应用
Abstract
(EN)
A glycoside conversion method based on high-speed countercurrent chromatography, relating to the technical field of glycoside conversion, and specifically relating to a use of a high-speed countercurrent chromatograph in the bioconversion of a glycoside component. In the present method, a high-speed countercurrent chromatograph serves as a reactor for performing a bio-enzyme conversion reaction on a glycoside component, differences in the distribution of the glycoside and an aglycone in two solvent phases are utilized, a glycoside hydrolase is added in advance to an aqueous phase solution as a stationary phase, and a glycoside substrate to be converted is added into a mobile phase. During operation of the countercurrent chromatograph, the enzyme is located in the stationary phase, and because the substrate glycoside more readily dissolves in the aqueous solution, in a mobile process, the glycoside enters the stationary phase from the mobile phase and undergoes a complete enzymolysis reaction with the glycoside hydrolase to produce a micromolecule aglycone, and the produced aglycone flows out with the mobile phase, completing the process of bioconversion and separation. In the present method, a high-speed countercurrent chromatograph serves as a bioconversion reactor, allowing conversion and separation to be completed at the same time, and allowing inhibiting effects to be eliminated from the enzymolysis reaction process, increasing the conversion rate.
(FR)
Procédé de conversion de glycoside basé sur une chromatographie à contre-courant à grande vitesse, et plus particulièrement, une utilisation d'un chromatographe à contre-courant à grande vitesse dans la bioconversion d'un composant glycoside. Dans la présente invention, un chromatographe à contre-courant à grande vitesse sert de réacteur pour effectuer une réaction de conversion de bio-enzyme sur un composant glycoside, des différences dans la distribution du glycoside et un aglycone dans deux phases de solvant sont utilisées, une hydrolase de glycoside est ajoutée à l'avance à une solution de phase aqueuse en tant que phase stationnaire, et un substrat de glycoside à convertir est ajouté dans une phase mobile. Pendant le fonctionnement du chromatographe à contre-courant, l'enzyme est située dans la phase stationnaire, et du fait que le glycoside de substrat se dissout plus facilement dans la solution aqueuse, dans un processus mobile, le glycoside entre dans la phase stationnaire à partir de la phase mobile et subit une réaction d'enzymolyse complète avec l'hydrolase de glycoside pour produire un aglycone de micromolécule, et l'aglycone produit s'écoule avec la phase mobile, achevant le processus de bioconversion et de séparation. Selon la présente invention, un chromatographe à contre-courant à grande vitesse sert de réacteur de bioconversion, permettant de réaliser la conversion et la séparation simultanément, et permettant d'éliminer les effets d'inhibition du processus de réaction d'enzymolyse, permettant ainsi d'augmenter le taux de conversion.
(ZH)
一种基于高速逆流色谱的糖苷转化方法,涉及糖苷转化技术领域,具体涉及高速逆流色谱仪在糖苷类成分生物转化中的应用。该方法将高速逆流色谱仪作为糖苷类成分进行生物酶转化反应的反应器,利用糖苷及苷元在两相溶剂之间的分配差异,预先将糖苷水解酶加入水相溶液中作为固定相,将待转化的糖苷底物加入流动相中。逆流色谱运转时,酶位于固定相中,由于底物糖苷更易溶于水溶液,在流动过程中会从流动相进入固定相与糖苷水解酶发生充分酶解反应生成小分子苷元,生成的苷元随流动相流出,完成生物转化及分离过程。该方法将高速逆流色谱仪作为生物转化反应器,能够一次性完成转化及分离,并且能够消除酶解反应过程中的抑制效应,增加转化率。
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