Method for using pulullan to prepare high-purity maltotriose

A technology of pullulan and maltotriose, which is applied in the preparation of sugar derivatives, chemical instruments and methods, oligosaccharides, etc., can solve the problems of not being able to be used as diagnostic reagents, difficulty in separation and extraction, and difficulty in extraction, etc. Low, high product purity, the effect of simplifying the preparation process

Inactive Publication Date: 2009-10-14
杨凌壹之农微生物工程技术研究院有限公司
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Problems solved by technology

[0003] However, the current production methods of maltotriose include: (1) high-quality syrup containing about 70% maltotriose is obtained by using wheat starch as raw material through enzymatic hydrolysis, which has low purity and is difficult to separate and extract, so it cannot be used as a diagnostic reagent; (2) Fermentation method, the reagent-grade high-purity maltotriose is obtained by fermentation, and there is also the problem of low purity and difficult extraction

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  • Method for using pulullan to prepare high-purity maltotriose

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Embodiment 1

[0018] (1) Dissolve 8kg of pullulan in 92kg of 0.2M pH5.7 phosphate buffer (dissolution and reaction are carried out in the same tank, the tank volume is 150L, ​​with stirring), and it is prepared to have a mass percent concentration of 8 % pullulan polysaccharide solution, then add 250g pullulanase, keep the temperature between 45~48°C, stir constantly, and the reaction time is 8 hours;

[0019] (2) After the reaction is over, the reaction liquid is pumped into the first storage tank, and then pumped into the ultrafiltration membrane separator by the first storage tank. The molecular weight cut-off of the membrane is 5000D, and the retained liquid is returned to the tank. Molecular salts and water permeate the membrane, and the permeate is collected in the second storage tank. During this process, deionized water is continuously added to the first storage tank. The amount of deionized water added is 50% of the liquid volume, when the residual amount of maltotriose in the rete...

Embodiment 2

[0023] (1) Dissolve 6.4kg of pullulan in 73.6kg of 0.2M pH5.7 phosphate buffer (dissolution and reaction are carried out in the same tank, the tank volume is 150L, ​​with stirring), and it is prepared as a mass percent concentration 8% pullulan polysaccharide solution, then add 200g pullulanase, keep the temperature between 45-48°C, keep stirring gently, and the reaction time is 8 hours;

[0024] (2) After the reaction is over, the reaction liquid is pumped into the first storage tank, and then pumped into the ultrafiltration membrane separator by the first storage tank. The molecular weight cut-off of the membrane is 7000D, and the retained liquid is returned to the tank. Molecular salts and water permeate the membrane, and the permeate is collected in the second storage tank. During this process, deionized water is continuously added to the first storage tank. The amount of deionized water added is 50% of the liquid volume, when the residual amount of maltotriose in the rete...

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Abstract

The invention discloses a method for using pulullan to prepare high-purity maltotriose. The technical proposal is as below: (1) using pullulanase to carry out zymohydrolysis on the pulullan; (2) using an ultrafiltration membrane to remove macromolecular protein type substances in reaction liquid; (3) using a nanofiltration membrane to remove micromolecular salt type substances and condense the reaction liquid; and (4) using chromatographic column separation to collect the pure product of the maltotriose, and obtaining the finished product of the maltotriose through vacuum drying. The method adopts an enzyme process to produce the maltotriose and realizes over 99 percent of product purity and low cost; the production process can realize continuity and automation; a great amount of pharmaceutical-grade maltotriose can be prepared; and the method is of great significance in producing a fast diagnosis reagent for curing acute pancreas diseases and developing saccharides with medicinal use.

Description

technical field [0001] The invention relates to a method for preparing maltotriose, in particular to a method for preparing high-purity maltotriose from pullulan. Background technique [0002] Maltotriose is a very valuable oligosaccharide composed of three glucose units connected by α-1,4-glucosidic bonds, and its molecular weight is 504.44. The determination of α-amylase has been used clinically for many years. Since the late 1970s, foreign countries have begun to develop methods for the determination of amylase with artificially synthesized substrates. In recent years, this type of research has developed rapidly and made great progress. It has been widely used, among which p-nitrophenol maltooligoglucoside (PNP-Gn) and 2-chloro-4-nitrophenol maltoside (CNP-Gn) are the main development trends, especially the substrates linked by maltotriose, because It does not need auxiliary enzymes, is cheap, and has high precision, and has been widely recognized and applied. Measureme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/16C12P19/00C07H3/06C07H1/06
Inventor 苏理赵双枝王金忠兰文忠蔺春兰董学前吉武科张彦昊魏连圣
Owner 杨凌壹之农微生物工程技术研究院有限公司
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