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Method for preparing α-cyclodextrin glucosyltransferase from marine microbial strain y112

A marine microorganism, Y112 technology, applied in the field of preparation of α-cyclodextrin glucosyltransferase, can solve the problems of unfavorable separation and purification of single cyclodextrin, limitation of cyclodextrin application, and uncommon use of chemical synthesis methods, etc.

Active Publication Date: 2018-05-22
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

[0003] At present, cyclodextrins are mainly prepared by enzymatic synthesis, chemical synthesis is not commonly used, and enzymatic preparation will produce a mixture of various cyclodextrins, which is not conducive to the separation and purification of a single cyclodextrin, which greatly limits Application of Cyclodextrin

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  • Method for preparing α-cyclodextrin glucosyltransferase from marine microbial strain y112

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Embodiment 1

[0033] Embodiment 1—preparation of α-CGTase by marine microbial strain Y112

[0034] 1. Preparation of crude enzyme solution

[0035] Put 30mL of seed culture medium in a 250mL Erlenmeyer flask to inoculate the marine microbial strain Y112, activate it at 30°C and 200r / min for 22h, and inoculate it into a Erlenmeyer flask containing 25mL of fermentation medium with an inoculation amount of 4% (V / V). (250mL), cultured at 27°C and 230r / min for 50h, collected the fermentation broth and centrifuged for 15min under a centrifugal force of 10000g, and the supernatant was the crude enzyme solution.

[0036] The composition of the seed medium: every liter of medium contains 10g of soluble starch, 5g of peptone, 5g of yeast extract powder, K 2 HPO 4 1g, MgSO 4 ·7H 2O 0.15g, NaCl 50g, sterilized at 121°C for 20min. The composition of the fermentation medium: every liter of medium contains 10g of cornstarch, 5g of peptone, 5g of yeast extract powder, K 2 HPO 4 1g, MgSO 4 ·7H 2 O ...

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Abstract

The invention relates to a method for preparing alpha-cyclodextrin glycosyltransferase by adopting a marine microbial strain Y112. The method comprises the following steps: preparing crude enzyme of the marine microbial strain Y112; carrying out fractional precipitation of ethanol; carrying out gel chromatography; and carrying out DEAE-Sepharose ion-exchange column chromatography. The prepared alpha-cyclodextrin glycosyltransferase product is high in single purity, the purity is improved by 12.4 times, and the recovery ratio is 57.5%. The alpha-cyclodextrin glycosyltransferase can be widely applied in the fields of medicine, food, chemical industry, cosmetics, industry, agriculture, and analytical chemistry.

Description

technical field [0001] The invention relates to the field of marine microorganisms, in particular to a method for preparing alpha-cyclodextrin glucosyltransferase from marine microorganism strain Y112. Background technique [0002] Cyclodextrin (cyclodextrin, abbreviated as CD) is formed by α-1,4-glucosidic linkages, cyclic maltooligosaccharides without reducing ends. Cyclodextrins usually consist of 6, 7 or 8 glucose residues and are called α-, β-, or γ-cyclodextrins, respectively. In addition to these three commonly used cyclodextrins, there are also δ-, ε-, ζ-, and η-cyclodextrins (composed of 9 to 12 glucose units, respectively). Cyclodextrins have the ability to encapsulate all or part of a variety of chemicals into cavities, thereby altering the physical and chemical properties of these compounds. Cyclodextrin is converted from starch by cyclodextrin glucosyltransferase (CGTase), and the main products are α-, β-, γ-cyclodextrin. Because the cyclodextrin separation a...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12R1/07
CPCC12N9/1074C12Y204/01019
Inventor 孙谧郝建华
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI