Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving oxidoreductase
Patent
1998-07-29
2000-03-07
Gitomer, Ralph
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving oxidoreductase
435 29, 435190, 435225, 4352561, C12Q 126, C12N 904, C12N 962, C12N 114
Patent
active
060338679
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a novel fructosyl amino acid oxidase. More particularly, it relates to a process for producing a fructosyl amino acid oxidase from the genus Aspergillus, a method of assaying an amadori compound using the enzyme, and a reagent or a kit containing the enzyme.
BACKGROUND ART
When reactive substances such as proteins, peptides and amino acids having an amino group(s) coexist with a reducing sugar such as aldose having an aldehyde group(s), they combine non-enzymatically and irreversibly through the amino and aldehyde groups, which is followed by the amadori rearrangement to form an amadori compound. The production rate of an amadori compound being a function of the concentration of reactants, contacting period, temperature, and the like, various useful information about a sample containing such a reactive substance(s) can be derived from the amount of amadori compounds. Examples of materials containing an amadori compound include food products such as soy sauce and body fluids such as blood.
In a living body, fructosylamines are formed through the glycation reaction between glucose and various amino acids. For example, fructosylamines produced when hemoglobin and albumin in blood are glycated are called glycohemoglobin and glycoalbumin, respectively. The reducing ability of glycated derivative of a protein in blood is called "fructosamine". The concentration of these glycated derivatives in blood reflects an average level of blood sugar over a given period of time, and the determination thereof can be a significant index for diagnosis and control of conditions of diabetes. Thercfore, the establishment of a method of measuring an amadori compound in blood must be clinically very useful. Further, the state of preservation and period after production of a food product can be estimated on the basis of the amount of amadori compounds in the food product, and, therefore, the method of measuring an amadori compound also contributes to the quality control of a food product. As mentioned above, an assay of amadori compounds should be useful in a wide range of fields involving medicine and food products.
Examples of assays of amadori compounds include a method which utilizes high performance liquid chromatography [Chromatogr. Sci. 10: 659 (1979)], a column filled with solid materials to which boric acid is attached [Clin. Chem. 28: 2088-2094 (1982)], electrophoresis [Clin. Chem. 26: 1598-1602 (1980)] or antigen-antibody reaction [JJCLA 18: 620 (1993), J. Clin. Lab. Inst. Reag. 16: 33-37 (1993)], a method for measuring the fructosamine [Clin. Chim. Acta 127: 87-95 (1982)], a calorimetric determination following the oxidization with thiobarbituric acid [Clin. Chim. Acta 112: 197-204 (1981)], or the like. These existing methods, however, require an expensive device(s) and are not necessarily accurate and rapid enough.
In the field of clinical assay and food analysis, a method utilizing enzymatic process has become more and more popular, which makes it possible to analyze an intended substance selectively with accuracy and rapidity owing to characteristics of enzymes (specificity in terms of substrate, reaction, structure, active site, etc.)
There have been provided assays for determining amadori compounds on the basis of the amount of oxygen consumed or hydrogen peroxide generated in the reaction between an amadori compound and an oxidoreductase (e.g. Japanese Patent Publication (KOKOKU) Nos. 5-33997 and 6-65300, and Japanese Patent Publication (KOKAI) Nos. 2-195900, 3-155780, 4-4874, 5-192193 and 6-46846). Further, assays of glycated protein for the diagnosis of diabetes have also been proposed (Japanese Patent Publication (KOKAI) Nos. 2-195899, 2-195900, 5-192193 (EP-A-0526150), 6-46846 (EP-A-0576838).
The reaction between an amadori compound and an oxidoreductase can be represented by the following general formula: --CO--CHO+R.sup.2 --NH.sub.2 +H.sub.2 O.sub.2 or peptide residue.
Examples of enzymes which catalyze the above reaction are as follows: Patent Publication
REFERENCES:
patent: 4985360 (1991-01-01), Takahashi et al.
patent: 5370990 (1994-12-01), Staniford et al.
patent: 5712138 (1998-01-01), Kato et al.
patent: 5789221 (1998-08-01), Kato et al.
Horiuchi et al. Purification and properties of fructosylamine oxidase from Aspergillus sp. 1005 Agric. Biol. Chem. 55(2), pp. 333-338 (Feb. 1991).
Yoshida et al. Distribution and properties of fructosyl amino acid oxidase in fungi. Appl. Environ. Microbiol. 61 (12), pp. 4487-4489. (1995).
Yoshida et al. Primary structures of fungal fructosyl amino acid oxidases and their application to the measurement of glycated proteins. EurJ. Biochem. 242 (3), pp. 499-505. (Dec. 15, 1996).
Cyousa Satoshi
Fukuya Hiroshi
Ishimaru Kaori
Kato Nobuo
Sakai Toshikatsu
Gitomer Ralph
Kyoto Dai-ichi Kagaku Co. Ltd.
Moran Marjorie A.
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