| 摘要 |
PURPOSE:To determine hydroxy-L-proline, specifically and easily, by reacting a liquid specimen with a hydroxy-L-proline dehydrogenation composite enzyme in the presence of a hydrogen-accepting pigment, and determining the color change of the pigment by optical means. CONSTITUTION:The amount of hydroxy-L-proline can be determined by reacting hydroxy-L-proline with a hydroxy-L-proline dehydrogenation composite enzyme in a proper solvent in the presence of a hydrogen-accepting pigment. The reaction is carried out under the condition to provide the above composite enzyme with high stability and the optimum activity. The solvent is water or especially a neutral or weakly basic buffer solution containing phosphoric acid, borate, etc. The hydrogen-accepting pigment is e.g. nitroblue tetrazolium, etc., and its amount is selected to be necessary and sufficient for the reaction. The reaction temperature is selected to provide the enzyme with high stability and optimum activity, and is preferably 30-37 deg.C.
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