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Method for preparing alpha-cyclodextrin glycosyltransferase by adopting marine microbial strain Y112

A marine microorganism, Y112 technology, applied in the field of preparation of α-cyclodextrin glucosyltransferase, can solve the problems of limiting the application of cyclodextrin, chemical synthesis method is not commonly used, unfavorable separation and purification of single cyclodextrin, etc.

Active Publication Date: 2015-11-25
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
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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 alpha-cyclodextrin glycosyltransferase by adopting marine microbial strain Y112

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Embodiment 1

[0033] Example 1 - Preparation of α-CGTase from marine microbial strain Y112

[0034] 1. Preparation of crude enzyme solution

[0035] After inoculating 30 mL of seed medium liquid in a 250 mL conical flask, inoculated with marine microbial strain Y112, activated at 30 °C and 200 r / min for 22 h, and inoculated into a conical flask containing 25 mL of fermentation medium at an inoculum of 4% (V / V) (250mL), cultured at 27°C and 230r / min for 50h, collected the fermentation broth and centrifuged for 15min under the condition of 10000g centrifugal force, and the supernatant was the crude enzyme liquid.

[0036] The composition of the seed medium: each liter of medium contains 10g of soluble starch, 5g of peptone, 5g of yeast extract, K 2 HPO 4 1g, MgSO 4 ·7H 2O0.15g, NaCl 50g, sterilized at 121°C for 20min. The composition of the fermentation medium: each liter of medium contains 10g of corn starch, 5g of peptone, 5g of yeast extract, K 2 HPO 4 1g, MgSO 4 ·7H 2 O0.15g, NaC...

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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 α-cyclodextrin glucosyltransferase from marine microorganism strain Y112. Background technique [0002] Cyclodextrin (CD for short) is a cyclic maltooligosaccharide with no reducing end linked by α-1,4-glycosidic bonds. Cyclodextrins typically consist of 6, 7 or 8 glucose residues and are referred to as alpha-, beta-, or gamma-cyclodextrins, respectively. In addition to these three commonly used cyclodextrins, there are also δ-, ε-, ζ- and η-cyclodextrins (respectively composed of 9 to 12 glucose units). Cyclodextrins have the ability to encapsulate a wide variety of chemicals in whole or in part within 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 separation and purifi...

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Application Information

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Patent Type & Authority Applications(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