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细胞色素 7A 对纤维素的水解表现出混合双曲线产物抑制。

Cellulose hydrolysis by cellobiohydrolase Cel7A shows mixed hyperbolic product inhibition.

机构信息

CITAB - Departamento de Biologia e Ambiente, Universidade de Trás-os-Montes e Alto Douro, Apartado 1013, 5001-801, Vila Real, Portugal.

出版信息

Appl Biochem Biotechnol. 2011 Sep;165(1):178-89. doi: 10.1007/s12010-011-9242-y. Epub 2011 Apr 16.

Abstract

In order to establish which are the contribution of linear (total), hyperbolic (partial) or parabolic inhibitions by cellobiose, and also a special case of substrate inhibition, the kinetics of cellobiohydrolase Cel7A obtained from Trichoderma reesei was investigated. Values of kinetic parameters were estimated employing integrated forms of Michaelis-Menten equations through the use of non-linear regression, and criteria for selecting inhibition models are discussed. With cellobiose added at the beginning of the reaction, it was found that cellulose hydrolysis follows a kinetic model, which takes into account a mixed hyperbolic inhibition, by cellobiose with the following parameter values: K (m) 5.0 mM, K (ic) 0.029 mM, K (iu) 1.1 mM, k (cat) 3.6 h(-1) and k (cat') 0.2 h(-1). Cellulose hydrolysis without initial cellobiose added also follows the same inhibition model with similar values (4.7, 0.029 and 1.5 mM and 3.2 and 0.2 h(-1), respectively). According to Akaike information criterion, more complex models that take into account substrate and parabolic inhibitions do not increase the modulation performance of cellulose hydrolysis.

摘要

为了确定纤维二糖的线性(总)、双曲(部分)或抛物抑制作用的贡献,以及底物抑制的特殊情况,研究了从里氏木霉中获得的纤维二糖水解酶 Cel7A 的动力学。通过使用非线性回归,利用米氏方程的积分形式来估计动力学参数值,并讨论了选择抑制模型的标准。在反应开始时添加纤维二糖时,发现纤维素水解遵循一个动力学模型,该模型考虑了纤维二糖的混合双曲抑制作用,其参数值如下:K(m) 5.0 mM,K(ic) 0.029 mM,K(iu) 1.1 mM,k(cat) 3.6 h(-1) 和 k(cat') 0.2 h(-1)。没有初始纤维二糖添加的纤维素水解也遵循相同的抑制模型,具有相似的值(分别为 4.7、0.029 和 1.5 mM 和 3.2 和 0.2 h(-1))。根据赤池信息量准则,考虑底物和抛物抑制作用的更复杂模型并没有增加纤维素水解的调制性能。

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