肺炎克雷伯菌4-羟基苯乙酸羟化酶编码基因的分子克隆与分析

Molecular cloning and analysis of the genes encoding the 4-hydroxyphenylacetate hydroxylase from Klebsiella pneumoniae.

作者信息

Gibello A, Suárez M, Allende J L, Martín M

机构信息

Departamento de Microbiología (Patología Animal I), Facultad de Veterinaria, Universidad Complutense, Avenida Puerta de Hierro s/n, 28040-Madrid, Spain.

出版信息

Arch Microbiol. 1997 Feb-Mar;167(2-3):160-6.

PMID:9133323

Abstract

The Klebsiella pneumoniae genes encoding the hydroxylase involved in the meta-cleavage pathway of 4-hydroxyphenylacetic acid (4-HPA) were cloned, and the DNA fragment from the region essential for hydroxylase activity was sequenced. K. pneumoniae 4-HPA hydroxylase was composed of two proteins (HpaA and HpaH) with different molecular masses. HpaA seems to be a flavin-containing hydroxylase with a molecular mass of 58,781 Da. HpaH, with a molecular mass of 18,680 Da, seems to be a "helper" protein required for productive hydroxylation of the substrate. The hpa genes were expressed and the hydroxylase was active in Escherichia coli. Comparison of the enzyme with other monooxygenases indicates that K. pneumoniae 4-HPA hydroxylase is a member of a new family of hydroxylases.

摘要

编码参与4-羟基苯乙酸（4-HPA）间位裂解途径的羟化酶的肺炎克雷伯菌基因被克隆，并对羟化酶活性所必需区域的DNA片段进行了测序。肺炎克雷伯菌4-HPA羟化酶由两种分子量不同的蛋白质（HpaA和HpaH）组成。HpaA似乎是一种分子量为58,781 Da的含黄素羟化酶。分子量为18,680 Da的HpaH似乎是底物有效羟化所需的“辅助”蛋白。hpa基因在大肠杆菌中表达且羟化酶具有活性。将该酶与其他单加氧酶进行比较表明，肺炎克雷伯菌4-HPA羟化酶是羟化酶新家族的成员。