ATYPICAL KINETICS OF IMMOBILIZED FIREFLY LUCIFERASE
ATYPICAL KINETICS OF IMMOBILIZED FIREFLY LUCIFERASE
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DOI：
10.1002/bit.260280804
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发表时间：
1986-08-01
影响因子：
3.8
通讯作者：
COULET, PR
中科院分区：
文献类型：
--
作者：
BLUM, LJ;COULET, PR
研究了胶原结合萤火虫荧光素酶的动力学性质。在反应介质中具有低搅拌的确定流体动力学条件下，与溶液中无酶相比，观察到的行为发生了改变：降低搅拌速率降低了观察到的酶活性。但是，扩散阻力不能单独解释这些非典型的动力学，虽然传质可能肯定发挥了重要作用，在生物发光反应的瞬态。固定化后，达到稳态所需的时间从300 ms增加到3 min，两种底物，胰蛋白酶和ATP，相对于酶的行为不同：首先与结合酶接触的饱和底物的性质并不是无关紧要的，这表明固定化可以揭示游离酶不可见的行为或机制。
The kinetic properties of collagen-bound firefly luciferase have been investigated. Under definite hydrodynamic conditions with low agitation in the reaction medium, the observed behavior is modified compared to the enzyme free in solution: reducing the stirring rate decreases the observed enzymatic activity. But diffusional resistances alone cannot account for these atypical kinetics though mass transfer may certainly play an important role during the transient state of the bioluminescent reaction. After immobilization, the time necessary to reach the steady state increased from 300 ms to 3 min and the two substrates, luciferin and ATP, behave differently with respect to the enzyme: The nature of the saturating substrate first in contact with the bound enzyme is not indifferent suggesting that immobilization can reveal behaviors or mechanisms which are not visualized with the free enzyme.