Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1994-08-08
1997-03-18
Wityshyn, Michael G.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435100, 435 84, 435 85, 435 95, 435 96, 435 98, 435 99, 435105, 536124, C12P 1904, C12P 1912, C12P 1914
Patent
active
056122039
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
This invention relates to a process for producing saccharides of definite chain length, such as glucose maltose, malto-oligosaccharides and isomalto-oligosaccharides with a high purity.
Among the saccharides of definite chain length which can be obtained by the process of the present invention, those having no branches include, inter alia, glucose and the species of the following formula: ##STR1## wherein m is an integer, m is preferably an integer of 0 to 5
Among such saccharides, maltose (m=0), maltotriose (m=1), maltotetraose (m=2), maltopentaose (m=3), maltohexaose (m=4) and the like can be used as pharmaceutical ingredients, food additives and starting materials for diagnostic reagents and so forth.
BACKGROUND OF THE INVENTION
Processes so far known for preparing high purity saccharides of definite in chain length generally comprise decomposing a saccharide having an arbitrary chain length, typically starch or the like, with one or more appropriate amylases and separating the object saccharide from the other unwanted oligo- and/or monosaccharides by the per se known column chromatographic and other fractionation techniques [cf. e.g. Jiro Nikuni (chief editor): Denpun Kagaku Handbook (Starch Science Handbook), published by Asakura Shoten, Tokyo, 1987, page 452] followed, if required, by crystallization of the object saccharide to further purify the same from the other unwanted contaminant oligo- and/or monosaccharides [cf. e.g. Denpun Kagaku Handbook (vide supra), page 456]
However, these processes invariably entail formation of unwanted byproduct oligosaccharides in addition to uncleaved dextrin and, therefore, are inherently disadvantageous.
As regards maltose, a process for producing high-purity maltose from terrestrial starch has been proposed (Japanese Kokai Tokkyo Koho JP 04-158795). However, this process is still unsatisfactory from the commercial viewpoint since high concentration charging of starch is impossible.
For solving these and other problems, there has been made an invention which is disclosed in the international patent application PCT/JP91/00984 (International Publication No. WO 92/01805). According to the above invention, a saccharide chain is transferred from a saccharide chain source, either directly or via an intermediate, to a substance which can be substantially separated from the desired saccharide of definite chain length (such substance is hereinafter referred to briefly as separable substance) by means of a transglycosidase, then the thus-obtained saccharide is treated with an enzyme capable of excising a saccharide chain having a specific chain length from said saccharide in an exo manner (such enzyme is hereinafter briefly referred to as exo-cleaving enzyme), and finally the desired saccharide of definite in chain length is isolated.
In this connection, a mode of causing an exo-cleaving enzyme to act on amylose is described by Nakakuki et al. [T. Nakakuki, K. Azuma and K. Kainuma, Carbohydrate Research, 128 (1984) 297-310] and the application of the periodate oxidation technique, among others, is described by Marshall et al. [J. J. Marschall and W. J. Whelan, Analytical Biochemistry, 43 (1971) 316-321]. However, these technologies have no direct relevance to the whole technology of the present invention.
Althogh the process described in PCT/JP91/00984 is an absolutely novel process capable of solving a number of problems, it still has the following disadvantages. accordingly a saccharide chain source must be provided for the supply of the necessary saccharide chain, the yield of the object saccharide is inevitably poor. have a structure such that it can serve as a receptor for the transglycosidase (for instance, a substance whose hydroxyl groups in positions 2, 3 and 4 have a glucose-type configuration, or ascorbic acid or the like). the action of an exo-cleaving enzyme, a procedure is essential for isolating said saccharide and, therefore, the yield is necessarily limited.
The object of the invention is to overcome the above technical drawbacks
REFERENCES:
patent: 4622295 (1986-11-01), Ikenaka et al.
patent: 4683298 (1987-07-01), Yalpani
patent: 4713118 (1987-12-01), Barker et al.
patent: 4787939 (1988-11-01), Barker
Kirk-Othmer Encyclopedia of Chemical Technology, vol. 13, 1981, pp. 678-705.
Robyt, et al, 145 Arch. Biochem and Biophysics 105-114 (1971).
Nakakuki, et al, 29 J. Jap. Soc. Starch Sci. 188-197 (1982).
Okemoto, et al, 25 Appl. Microbiol Biotechnol 137-142 (1986).
Ezure Yohji
Maruo Shigeaki
Tachikake Noriyuke
Nippon Shinyaku Co. Ltd.
Prats Francisco C.
Wityshyn Michael G.
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