| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2000: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
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| Research Abstract |
CHEMILUMINOMETRIC FLOW-THROUGH SENSORS WITH IMMOBILIZED ENZYMES IN FLOW INJECTION SYSTEMS HAVE BEEN DEVELOPED. PEROXIDASE." LEUCINE DEHYDROGENASE, NADH OXIDASE, FORMALDEHYDE DEHYDROGENASE, URICASE, GLUTAMATE OXIDASE, AND LYSINE OXIDASE WERE IMMOBILIZED COVALENTLY ON TRESYLATED HYDROPHILIC VINYL POLYMER BEADS AND PACKED INTO TRANSPARENT PTFE TUBE (1.0 MM I.D. X 1.5 MM O.D.), AND THE TUBE WAS PLACED IN FRONT OF A PHOTOMULTIPLIER TUBE AS A FLOW CELL THE STABILITIES OF THE IMMOBILIZED ENZYMES WERE SUPERIOR TO THOSE PREVIOUSLY USED, BECAUSE THESE WERE THERMOPHILIC ENZYMES. THE SENSORS WERE STABLE FOR A MONTH. IT WAS FOUND THAT THE PEROXIDASE FROM ARTHROMYCES RAMOSUS IS WELL SUITED AS CATALYST FOR CHEMILUMINOMETRIC FLOW-INJECTION MEASUREMENTS OF LUMINOL-HYDROGEN PEROXIDE REACTION.
A SELECTIVE FLOW-THROUGH SENSOR FOR THE DETERMINATION OF L-GLUTAMATE IN SERUM WAS DEVELOPED. THE PRINCIPLE FOR THE SELECTIVE DETECTION FOR L-GLUTAMATE WAS BASED ON COUPLED RACTIONS OF FOUR SEQUENTIALLY ALIGNED IMMOB
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ILIZED OXJDASES, UIRCASE/PEROIXIDASE/ GLUTAMATE OXIDASE/PEROXIDASE IN A FLOW CELL THE IMMOBILIZED URICASE WAS USED EMPLOYED TO DECOMPOSE URATE, WHICH IS ONE OF THE MAJOR INTERFERING COMPONENTS IN SERUM FOR LUMINOL-HYDROGEN PEROXIDE CHEMILUMINESCENCE REACTION. THE HYDROGEN PEROXIDE PRODUCED FROM THE URICASE REACTION READILY REACTED WITH REDUCING COMPONENTS, SUCH AS ASCORBATE AND GLUTATHIONE, AND THEN THE EXCESS HYDROGEN PEROXIDE WAS DECOMPOSED BY THE IMMOBILIZED PEROXIDASE. L-GLUTAMATE IN THE SAMPLE PLUG WAS ENZYMATICALLY CONVERTED TO HYDROGEN PEROXIDE WITH IMMOBILIZED GLUTAMATE OXIDASE SUBSEQUENTLY THE PEROXIDE REACTED WITH LUMINOL ON THE IMMOBILIZED PEROXIDASE TO PRODUCE CHEMILUMINESCENCE, PROPORTIONAL TO GLUTAMATE CONCENTRATION.
A FLOW-THROUGH SENSOR FOR THE DETERMINATION OF HISTAMINE IN FISH MEAT WAS DEVELOPED. HISTAMINE OXIDASE WAS FOUND IN CELLS OF ARTHROBACTER CRYSTALLOPOITES KAIT-B-007 ISOLATED FROM SOIL THE OXIDASE AND PEROXIDASE WERE COIMMOBILIZED COVALENTLY ON POLYMER BEADS AND PACKED INTO THE TPTFE TUBE. ONE ASSAY FOR HISTAMINE WAS DONE AT INTERVALS OF 2 MIN WITHOUT CARRYOVER. THE RESPONSE WAS REPRODUCIBLE WITHIN 1.25 % OF THE RELATIVE STANDARD DEVIATION FRO ll-REPLICATE INJECTIONS OF 50 MM HISTAMINE. Less
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