1997 Fiscal Year Final Research Report Summary
Design of Molecular-signal Responsive Enzyme Systems Using Bioaffinity Binding and Dissociation
| Project/Area Number |
08650947
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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 |
生物・生体工学
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| Research Institution | Okayama University |
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Principal Investigator |
SHINOHARA Hiroaki Okayama Univ.Faculty of Eng.Associate Professor, 工学部, 助教授 (60178887)
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| Project Period (FY) |
1996 – 1997
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| Keywords | bioaffinity binding / enzyme activity / ligand-modification / biotin-avidin / molecular signal / synthetic coenzyme / apo enzyme / intelligent molecular devise |
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| Research Abstract |
Recently development of intelligent molecular system is strongly requested. The author have paied much attention to allosteric enzyme moleules in which the enzyme activity is controlled by the molecular-signal responsive conjugate formation with catalytic protein arid regulatory protein. In this research, molecular signal-responsive enzymatic reaction systems was desingned and constructed by using bioaffinity binding and dissociation between avidin molecule and biotin- or biotin analogue-modified enzyme.
In the first year, HABA (hydroxybenzeneazo-benzoic acid)-modified beta-Galactosidase was prepared as a catalytic site and conjugated with avidin as a regulatory site. The beta-Galactosidase activity was suppressed at about 60% after conjugate formation and was recovered almost completely by free biotin addition at the sutable modification ratio.
In the second year, biotin-modified flavin coenzyme derivatives were synthesized and reconstituted into apo-D-amino acid oxidase. A biotin-binding protein, avidin bound to the reconstituted enzyme and it resulted in the enzyme activity suppression. Following addition of free biotin induced dissociation of the enzyme-avidin complexs and resulted in a partial recovery of the enzyme activity.
In conclusion, the author would like to say the direction to develope intelligent enzyme molecular systems in which the enzymatic activity is regulated by molecular signal was open.
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