来自苯甲酸降解型不透明红球菌1CP的底物/底物类似物与苯甲酸1,2-双加氧酶之间的相互作用动力学

Kinetics of interaction between substrates/substrate analogs and benzoate 1,2-dioxygenase from benzoate-degrading Rhodococcus opacus 1CP.

作者信息

Solyanikova Inna P, Borzova Oksana V, Emelyanova Elena V

机构信息

FSBIS G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, prospect Nauki, 5, Pushchino, Moscow region, 142290, Russia.

Pushchino State Natural Science Institute, Pushchino, Russia.

出版信息

Folia Microbiol (Praha). 2017 Jul;62(4):355-362. doi: 10.1007/s12223-017-0505-z. Epub 2017 Feb 24.

DOI:10.1007/s12223-017-0505-z

PMID:28236176

Abstract

Benzoate 1,2-dioxygenase (BDO) of Rhodococcus opacus 1CP, which carried out the initial attack on benzoate, was earlier shown to be the enzyme with a narrow substrate specificity. A kinetics of interaction between benzoate 1,2-dioxygenase and substituted benzoates was assessed taking into account the enlarged list of the type of inhibition and using whole cells grown on benzoate. The type of inhibition was determined and the constants of a reaction of BDO with benzoate in the presence of 2-chlorobenzoate (2CBA), 3,5-dichlorobenzoate (3,5DCBA), and 3-methylbenzoate (3MBA) were calculated. For 2CBA and 3MBA, the types of inhibition were classified as biparametrically disсoordinated inhibition and transient inhibition (from activation towards inhibition), respectively. The process of not widely recognized pseudoinhibition of a BDO reaction with benzoate by 3,5DCBA was assessed by the vector method for the representation of enzymatic reactions. Ki value was determined for 2CBA, 3MBA, and 3,5DCBA as 337.5, 870.3, and 14.7 μM, respectively.

摘要

对苯二甲酸1,2-双加氧酶（BDO）来自红平红球菌1CP，它对苯甲酸进行初始攻击，早期研究表明该酶具有狭窄的底物特异性。考虑到抑制类型列表的扩充，并使用在苯甲酸上生长的全细胞，评估了苯甲酸1,2-双加氧酶与取代苯甲酸之间的相互作用动力学。确定了抑制类型，并计算了BDO在2-氯苯甲酸（2CBA）、3,5-二氯苯甲酸（3,5DCBA）和3-甲基苯甲酸（3MBA）存在下与苯甲酸反应的常数。对于2CBA和3MBA，抑制类型分别分类为双参数不协调抑制和瞬时抑制（从激活到抑制）。通过用于表示酶促反应的向量法评估了3,5DCBA对BDO与苯甲酸反应的未被广泛认可的假抑制过程。确定2CBA、3MBA和3,5DCBA的Ki值分别为337.5、870.3和14.7μM。