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A mild intensity of the enzyme-support multi-point attachment promotes the optimal stabilization of mesophilic multimeric enzymes: Amine oxidase from Pisum sativum.
Journal of Biotechnology ( IF 4.1 ) Pub Date : 2020-05-12 , DOI: 10.1016/j.jbiotec.2020.04.006
Paz García-García 1 , Jose M Guisan 2 , Gloria Fernandez-Lorente 1
Affiliation  

Stabilization of dimeric enzymes requires the stabilization of the quaternary structure as well as the 3D one. Both subunits may be easily immobilized on a highly activated support. Additional stabilization of the 3D structure may be achieved via multipoint covalent attachment (MCA) on highly activated supports. In the case of monomeric enzymes or thermophilic dimeric ones, the optimal stabilization is obtained via the most intense MCA and it is associated to a small loss of catalytic activity. However, in the case of mesophilic enzymes, a very intense MCA of both subunits may promote negative effects, e.g., associated to distortions of the assembly between subunits and a subsequent very important loss of catalytic activity. A dimeric mesophilic amine oxidase from P.sativum was stabilized by MCA on glyoxyl-agarose. Both subunits were covalently immobilized on the support through the region with the highest density in Lys residues. In addition to that, an interesting activity/stabilization binomial was obtained after only 3 h of enzyme-support multiinteraction (50 % of activity/350 fold stabilization). However, after 24 h of enzyme-support multi-interaction this binomial activity-stabilization decreased down to 30/150. A moderate multiinteraction seems to be the optimal strategy for immobilization-stabilization of mesophilic dimeric enzymes and it promotes moderate losses of activity and interesting stabilizations against the combined effect of heat, acid pH and ethanol. The control of the intensity of enzyme-support multi-interactions becomes now strictly necessary.

中文翻译:

中等强度的酶支持多点连接可促进中温多聚酶(Pisum sativum的胺氧化酶)的最佳稳定性。

二聚体酶的稳定化需要四级结构和3D稳定化。两个亚基都可以容易地固定在高度活化的载体上。3D结构的其他稳定性可以通过在高度活化的支持物上进行多点共价连接(MCA)来实现。在单体酶或嗜热二聚酶的情况下,可通过最强的MCA获得最佳的稳定性,这与催化活性的少量损失有关。但是,在嗜温酶的情况下,两个亚基的非常强烈的MCA可能会促进负面影响,例如,与亚基之间组装的变形以及随后催化活性的非常重要的损失相关。MCA在乙醛琼脂糖上稳定了来自腐霉的二聚体嗜温胺氧化酶。通过在Lys残基中具有最高密度的区域将两个亚基共价固定在支持物上。除此之外,仅3小时的酶支持多重相互作用(50%的活性/ 350倍的稳定性)就获得了令人感兴趣的活性/稳定性二项式。但是,经过24小时的酶支持多相互作用后,该二项式活动稳定度下降至30/150。适度的多重相互作用似乎是嗜温二聚体酶固定化-稳定化的最佳策略,它促进了适度的活性丧失和对热,酸性pH和乙醇的综合作用产生了令人感兴趣的稳定作用。现在严格地需要控制酶-载体多重相互作用的强度。除此之外,仅3小时的酶支持多重相互作用(50%的活性/ 350倍的稳定性)就获得了令人感兴趣的活性/稳定二项式。但是,经过24小时的酶支持多相互作用后,该二项式活动稳定度下降至30/150。适度的多重相互作用似乎是嗜温二聚体酶固定化-稳定化的最佳策略,它促进了适度的活性损失和对热,酸性pH和乙醇综合作用的稳定作用。现在严格地需要控制酶-载体多重相互作用的强度。除此之外,仅3小时的酶支持多重相互作用(50%的活性/ 350倍的稳定性)就获得了令人感兴趣的活性/稳定性二项式。但是,经过24小时的酶支持多相互作用后,该二项式活动稳定度下降至30/150。适度的多重相互作用似乎是嗜温二聚体酶固定化-稳定化的最佳策略,它促进了适度的活性丧失和对热,酸性pH和乙醇的综合作用产生了令人感兴趣的稳定作用。现在严格地需要控制酶-载体多重相互作用的强度。在24小时的酶支持多相互作用后,该二项式活动稳定度降低至30/150。适度的多重相互作用似乎是嗜温二聚体酶固定化-稳定化的最佳策略,它促进了适度的活性丧失和对热,酸性pH和乙醇的综合作用产生了令人感兴趣的稳定作用。现在严格地需要控制酶-载体多重相互作用的强度。经过24小时的酶支持多相互作用后,该二项式活动稳定度降低至30/150。适度的多重相互作用似乎是嗜温二聚体酶固定化-稳定化的最佳策略,它促进了适度的活性丧失和对热,酸性pH和乙醇的综合作用产生了令人感兴趣的稳定作用。现在严格地需要控制酶-载体多重相互作用的强度。
更新日期:2020-05-12
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