Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1993-02-04
1994-11-15
Lilling, Herbert J.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435 97, 435 98, 435 99, 435101, 536 41, 536 174, 536 185, 536 186, C12P 1920, C12P 1918, C12P 1902, C12P 1914
Patent
active
053647947
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a process for producing saccharides of definite chain length, such as glucose, maltose and maltooligosaccharides, each in an isolated and highly pure form.
BACKGROUND ART
Processes so far known for producing saccharides of definite chain length each in an isolated and highly pure form comprise hydrolyzing a glucan, such as starch, with one or more appropriate amylases and separating the desired saccharide from other unwanted oligosaccharides and/or monosaccharides by some or other known column chromatographic technique or the like [e.g. Denpun Kagaku Handbook (Starch Science Handbook), p. 452, 1987; Japanese Kokai Tokkyo Koho JP 57-209000; Japanese Kokai Tokkyo Koho JP 62-19210; Japanese Patent Publication No. 02-17158], and/or crystallizing the desired saccharide to thereby separate the same from other unwanted oligosaccharides and/or monosaccharides occurring in trace quantities (e.g. Denpun Kagaku Handbook, p. 456, 1987).
However, all the processes mentioned above intrinsically entail formation of such byproducts as glucose, uncleaved dextrin and other contaminant oligosaccharides and substantially fail to remove these unwanted saccharides from the reaction mixture.
It is known that the coexistence of an unwanted saccharide or saccharides in trace amounts markedly interfere with crystallization of the desired carbohydrate of definite chain length.
Furthermore, when the known processes mentioned above is employed for producing an oligosaccharide in a purity as high as possible, the production procedure becomes complicated, presenting problems from the yield and cost points of view.
DISCLOSURE OF INVENTION
Accordingly, the inventors of the present invention considered that these known production processes for saccharides have much to be improved and attempted to establish a process for producing a saccharide of definite chain length, such as glucose, maltose, or a maltooligosaccharide, in a form substantially free of unwanted saccharides, so that it may be suited for pharmaceutical use.
The gist of the invention lies in a serial and continuous execution of the following procedures.
Thus, the process of the invention comprises causing a saccharide chain to be transferred from a saccharide source, either directly or via an intermediate, to a substance substantially separable from the desired saccharide (hereinafter referred to as "separable substance") using a saccharide chain transferase, treating the resulting oligosaccharide with an enzyme capable of excising a saccharide chain of definite chain length therefrom in an exo manner (hereinafter referred to as "exo-cleaving enzyme") and isolating the desired saccharide of definite chain length.
In the following, the invention is described in detail.
The separable substance to be used in accordance with the invention is a carrier or support of the type currently in use as an immobilization carrier, such as chitosan beads, an ion exchange resin, a synthetic resin or the like, or a compound of the general formula [I] ##STR1## (wherein R is hydrogen, lower alkyl, hydroxyalkyl, phenylalkyl, phenylalkenyl, phenylalkynyl, phenoxyalkyl, phenoxyalkenyl or phenoxyalkynyl, including the case where the phenyl moiety is substituted) (herein-after referred to as "a moranoline"), nojirimycin, an aminocyclitol, an aminocyclitol derivative, glucuronic acid, a glucosylated or oliogoglucosylated modification thereof or the like polar saccharide adsorbable on an ion exchange resin (hereinafter collectively referred to as "polar sugar"), or the like. The carrier may be in any form, e.g. bead-like, membrane-like or fibrous, for instance. Moranolines can be obtained from the mulberry white rind, actinomycetes and so on (e.g. Japanese Patent Application No. 54-159417, Japanese Patent Application No. 55-76838, Japanese Patent Publication No. 56-9919, Japanese Patent Application No. 57-93997, Japanese Patent Publication No. 59-27337). Glucosylated or oliogoglucosylated moranolines can be produced by the methods already disclosed [e.g.
REFERENCES:
patent: 3878191 (1975-04-01), Fukumoto et al.
patent: 5256788 (1993-10-01), Ezure et al.
Ezure Yohji
Maruo Shigeaki
Miyazaki Katsunori
Yamada Naoyoshi
Lilling Herbert J.
Nippon Shinyaku Company Limited
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