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Method for preparing straight-chain maltopentaose by using double-enzyme method

A technology of maltopentasaccharide and maltooligosaccharides, which is applied in the direction of medical preparations, applications, and pharmaceutical formulations containing active ingredients, which can solve the problems of low substrate concentration and inability to directly expand industrial production, and reduce production and processing costs , good thermal stability and simple process

Active Publication Date: 2018-07-20
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On domestic and international laboratory research levels, the substrate concentration used is all low (1~5%), and conversion rate level is generally about 50%~75%; The content of maltopentaose is only qualitatively analyzed without quantitative explanation, so it cannot be directly expanded and applied to industrial production

Method used

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  • Method for preparing straight-chain maltopentaose by using double-enzyme method
  • Method for preparing straight-chain maltopentaose by using double-enzyme method
  • Method for preparing straight-chain maltopentaose by using double-enzyme method

Examples

Experimental program
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Effect test

Embodiment 1

[0029] A 10% maltodextrin (DE=6) solution with pH 6.0 was prepared as a substrate, and the malto-oligosaccharide generating enzyme (the amino acid sequence numbered in GenBank is AIV43245.1) was added at a rate of 100 U / g with the enzyme added, and 2 U / g g Add the amount of enzyme and add pullulanase, react at 60°C for 24 to 72 hours. After the reaction, the enzyme is inactivated in a boiling water bath for 40 minutes, and the sample is measured by high performance anion exchange chromatography-pulse amperometric detection (HPAEC-PAD). The chromatogram of 24h reaction is as follows figure 1 Shown. The relative content of each component in the syrup corresponding to different reaction times is as follows figure 2 As shown, at this time, the main product maltopentaose content is above 43%, the substrate conversion rate is 90.2%-99.8%, and the glucose content is within 10%, and the linear oligosaccharides with a DP value of 8 and above are not detected. It shows that the substrat...

Embodiment 2

[0031] Prepare 30% maltodextrin (DE=6) solution as substrate, adjust pH 5.5, add malto-oligosaccharide-producing enzyme (amino acid sequence number in GenBank is AIV43245.1) according to 50U / g enzyme amount, press Pullulanase was added with 5U / g enzyme amount, reacted at 70℃ for 72h, after the reaction, the enzyme was killed in a boiling water bath, and the composition and content of the sample were determined by high performance anion exchange chromatography-pulse amperometric detection (HPAEC-PAD) Such as image 3 As shown, at this time, the main product maltopentaose content in the syrup is 40.5%, and the substrate conversion rate reaches 91.7%.

Embodiment 3

[0033] Prepare a 10% corn starch solution as the substrate, adjust the pH to 6.5, add the malto-oligosaccharide generating enzyme (amino acid sequence number in GenBank is AIV43245.1) according to the 100U / g enzyme amount, and add the enzyme amount according to 2U / g Add pullulanase and react at 60℃ for 48h. After the reaction, the enzyme is killed in a boiling water bath. The content of the main product maltopentaose in the syrup is 43.3% as determined by high performance anion exchange chromatography-pulse amperometric detection (HPAEC-PAD). The substrate conversion rate reached 96.5%.

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Abstract

The invention relates to a method for preparing straight-chain maltopentaose by using a double-enzyme method, and belongs to the technical field of the production of functional sugar. The number, in the GenBank, of the amino acid sequence of a used straight-chain maltooligosaccharide producing enzyme is AIV43245.1; the used pullulanase is purchased from the Japanese Amano Enzyme Inc.. The method comprises the following steps of preparing starch or maltodextrin solution with the pH (potential of Hydrogen) of 5.5 to 6.5 as a substrate, adding the straight-chain maltooligosaccharide producing enzyme according to an enzyme addition amount of 50U / g to 100U / g, adding the pullulanase according to an enzyme addition amount of 2U / g to 5U / g, and reacting for 24 to 72 hours at 60 to 70 DEG C to subsequently obtain straight-chain maltooligosaccharide syrup, wherein a conversion ratio (measured according to glucose to straight-chain maltoheptaose) reaches 90 percent or above; a main product is thestraight-chain maltopentaose; the percent of the main product can reach 40 percent or above. Two types of enzymes used by the method can be simultaneously added; the reaction temperature or the pH does not need to be regulated midway; the alpha-amylase is also not needed to liquefy the substrate; a calcium ion does not need to be added, and the method is simple in production process, is safe and further economical, and has higher application value.

Description

Technical field [0001] The invention relates to a method for preparing linear maltopentaose by a double-enzyme method, and belongs to the technical field of functional sugar production. Background technique [0002] Linear malto-oligosaccharides refer to a class of functional oligosaccharides composed of 3-10 glucose units through α-1,4-glycosidic bonds. It has good food processing adaptability: it can be used as a soft taste sweetener; it can be used as a moisture regulator in baked and puffed foods; it can improve the melting resistance of cold drinks and increase the expansion rate of ice cream in cold drinks; in chocolate, It can effectively inhibit crystal crystallization in foods such as jam; it can be used as a thickener in liquid foods; in addition, it can inhibit starch aging and protein denaturation in quick-frozen foods, thereby extending the shelf life. [0003] Straight-chain malto-oligosaccharides also have the following unique physiological effects: they are digeste...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14A23L33/125A23L29/30A61K31/702A61P3/02
CPCA23V2002/00A61K31/702A23L29/30A23L33/125C12P19/14A23V2200/32A23V2200/30A23V2250/282
Inventor 李兆丰顾正彪潘思惠李才明王颖兰程力洪雁
Owner JIANGNAN UNIV
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