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CN-121991142-A - Method for preparing trehalose by utilizing purple tremella

CN121991142ACN 121991142 ACN121991142 ACN 121991142ACN-121991142-A

Abstract

The invention discloses a method for preparing trehalose by utilizing purple fungus, which is characterized in that the purple fungus strain is high-temperature type edible fungus and is preserved in the Guangdong province microorganism strain preservation center (GDMCC) with the preservation number of GDMCC NO:62288, the method comprises the following steps of (1) leaching smashed purple fungus fruiting bodies by adopting an organic solvent, concentrating the obtained leaching liquid, sequentially carrying out liquid-liquid extraction by using ethyl acetate, water and n-butanol, collecting n-butanol phase, and carrying out dehydration concentration to obtain an organic crude extract, and (2) dissolving the extract obtained in the step (1), and sequentially purifying by reverse phase silica gel column chromatography, gel exclusion column chromatography and normal phase silica gel column chromatography to obtain the trehalose. The extraction and purification process provided by the invention does not need complex enzyme catalysis or fermentation steps, and avoids the use of expensive enzyme preparations.

Inventors

  • YUAN BIN
  • KE LINA
  • ZHENG YONGBIAO
  • LIAN YANPING
  • Bai Cuie
  • CHEN TIANCI
  • WU ZHENQIANG

Assignees

  • 漳州市农业科学研究所(漳州市茶叶科学研究所)

Dates

Publication Date
20260508
Application Date
20251211

Claims (8)

  1. 1. The application of the purple fungus fruiting body in preparing trehalose, wherein the purple fungus strain is a high-temperature type edible fungus which is preserved in the Guangdong province microorganism strain preservation center GDMCC, and the preservation number is GDMCC NO: 62288.
  2. 2. A method for preparing trehalose using a fruiting body of a strain of purple ear according to claim 1, comprising the steps of: (1) Leaching the crushed purple eared fruiting body by adopting an organic solvent, concentrating the obtained leaching solution, sequentially carrying out liquid-liquid extraction by using ethyl acetate, water and n-butanol, collecting n-butanol phase, and dehydrating and concentrating to obtain an organic crude extract; (2) And (3) dissolving the extract obtained in the step (1), and purifying by reverse phase silica gel column chromatography, gel exclusion column chromatography and normal phase silica gel column chromatography in sequence to obtain the trehalose.
  3. 3. The method according to claim 2, wherein the organic solvent in step (1) is selected from one or more of water, methanol, ethanol, acetone, ethyl acetate.
  4. 4. The method according to claim 2, wherein the reversed-phase silica gel column chromatography in step (2) is gradient-eluted with a methanol-water solution.
  5. 5. The method according to claim 2, wherein the gel exclusion column chromatography in step (2) is performed using a Sephadex LH-20 gel column eluting with methanol as a mobile phase.
  6. 6. The method according to claim 2, wherein the normal phase silica gel column chromatography in step (2) is eluted with a chloroform-methanol mixed solution.
  7. 7. A high purity trehalose prepared by the process of any one of claims 2 to 6.
  8. 8. A trehalose obtained by the method according to claim 2, wherein the trehalose has a molecular formula of C12H12O11 and a structural formula of: 。

Description

Method for preparing trehalose by utilizing purple tremella Technical Field The invention belongs to the field of biotechnology and natural product separation and extraction, and particularly relates to a specific purple fungus (Auricularia thailandica) strain and application thereof in preparing trehalose, in particular to a method for efficiently separating and purifying high-purity trehalose from purple fungus fruiting bodies. Background Trehalose (Trehalose) is a non-reducing disaccharide composed of two molecules of glucose through alpha, alpha-1, 1-glycosidic linkages. The unique biological functions are realized in that the biological functions have remarkable protective effects on biological macromolecules (such as proteins and liposomes) and cell structures under the adverse conditions of high temperature, freezing, drying, high permeability and the like, so that the biological functions are widely applied to the food industry (serving as a natural preservative and a stabilizer), the medicine field (serving as auxiliary materials of vaccines, enzyme preparations and cytoprotective agents) and high-end cosmetics, and are known as 'life sugar'. Currently, commercial production of trehalose relies mainly on microbial fermentation and enzymatic conversion. Enzymatic conversion usually takes starch or maltose as a substrate, and utilizes the synergistic effect of maltooligosaccharide-based trehalose synthase and maltooligosaccharide-based trehalose hydrolase, or utilizes trehalose synthase for conversion. Although the enzyme-producing strain is continuously optimized by genetic engineering, immobilized enzyme technology is adopted, and the like, the process has the inherent limitations that (1) the cost of raw materials is obviously influenced by grain price fluctuation, (2) the cost of enzyme preparations is high, (3) the subsequent separation and purification steps are complicated, so that residual substrates, byproducts and enzyme proteins are removed, and the overall cost is further increased. Therefore, development of new raw materials and new processes which do not depend on grain matrixes, are simple in process and low in cost has become an important research direction in the field. Disclosure of Invention The purple fungus (Auricularia thailandica) is a kind of edible and medicinal fungus with abundant resources. However, the prior art does not relate to a method for specific extraction and purification of trehalose directly from the purple-eared fruit body. The purple tremella is excavated as a new raw material source for producing trehalose, so that the dependence on the traditional grain matrix can be effectively reduced, and a new direction is provided for developing high-added-value fungus products. The invention is based on a specific preserved purple fungus strain, the fruiting body of the purple fungus strain is found to be rich in trehalose, and corresponding extraction and purification technology is developed accordingly, so that high-purity trehalose is successfully separated. The method provides a brand new non-grain raw material path solution for the production of trehalose. The primary aim of the invention is to overcome the defects of the prior art and provide a strain of purple fungus (Auricularia thailandica) which can be used for preparing trehalose. And further provides a method for preparing trehalose by utilizing the purple otocarpal fruit bodies, which has the advantages of simple process, low cost and high product purity. The strain of purple fungus (Auricularia thailandica) of the invention is 3 months and 10 days in 2022. Is deposited with the microorganism strain collection (GDMCC) of Guangdong province with the deposit number of GDMCC NO: 62288. The invention also provides a method for preparing trehalose by utilizing the fruiting body of the purple fungus strain, which comprises the following steps: (1) The organic leaching comprises the steps of crushing dried purple otocarpus fruit bodies, leaching by adopting an organic solvent, collecting leaching liquor and concentrating to obtain concentrated liquor, carrying out equal volume extraction on the concentrated liquor by adopting ethyl acetate and water, retaining a water phase, carrying out equal volume extraction on the water phase by adopting n-butanol, retaining a n-butanol phase, and dehydrating and concentrating the n-butanol phase to obtain an organic crude extract. (2) And (3) separating and purifying, namely dissolving the organic crude extract obtained in the step (1) by using methanol, sequentially carrying out fractional purification on the obtained sample solution by reverse phase silica gel column chromatography, gel exclusion column chromatography and normal phase silica gel column chromatography, collecting the component rich in trehalose, concentrating and drying to obtain a pure product of the trehalose. Preferably, in the step (1), the organic solvent is one or more mixed solvents of water, methano