| Project/Area Number |
09680060
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
家政学
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| Research Institution | KACHO JUNIOR COLLEGE |
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Principal Investigator |
TOYOHARA Masako HUMAN LIFE STUDIES, KACHO JUNIOR COLLEGE, ASSOCIATE PROFF., 生活学科, 助教授 (50241211)
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| Co-Investigator(Kenkyū-buntansha) |
NAKATA Reiko NARA WOMEN'S UNIV., FAC. HUMAN LIFE & ENVIR., LECTURER, 生活環境学部, 講師 (90198119)
TOYOHARA Haruhiko KYOTO UNIV., APPLIED BIOSCI, ASSOCIATE FROFF., 大学院・農学研究科, 助教授 (90183079)
MURATA Michiyo HUMAN LIFE STUDIES, KACHO JUNIOR COLLEGE, PROFF., 生活学科, 教授 (30133135)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | FIBRYNOLYTIC ENZYME / MUSHROOM / FIBRINOGEN ZYMOGRAPHY / 血栓溶解酵素 |
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| Research Abstract |
The purpose of the present study was to search fibrinolytic activities among various food materials by using the fibrinolytic assay and fibrinogen zymography originally devised by the authors. The author also made attempts to examine the intake of the active enzyme in the blood of animal by oral administration and the reduction of the activity during curing.
Among more than 200 materials examined, the author successfully detected the strong activities in mushrooms in addition to natto as reported. Judged from the inhibitory effect by 1,10-phenanthroline, most of the activities from mushrooms were supposed to be derived from metalloproteinases. The most heat stable activity was found in edible mushroom basidiomycete (Grifola frondosa), which remained after short period heating at 90℃. Saute was shown to be the most suitable cooking to reserve the activity.
Purification of the enzyme by conventional ion exchange chromatographies from the fruiting body of basidiomycete was hampered possibly because the enzyme formed a complex with glycoproteins with various isoelectric points. By gel permeation chromatographies, the enzyme molecule was eluted as a relatively low molecular weight molecule (approximately 20kDa). The intake of the enzyme into blood by oral administration was examined by using antibodies raised against whole extract of basidiomycete, but significant intake was not immunologically detected. The cDNA possibly encoding the enzyme was cloned by PCR method using primers designed form the report determining the amino acid sequence of the enzyme (J. Biol. Chem., 272, 30032-30039(1999)). The successful cloning of the enzyme would lead the bacterial production of the large amount of the enzyme as recombinant proteins.
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