Kato Yasuo, Tsuda Takashi, Asano Yasuhisa
Biotechnology Research Center, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
Biochim Biophys Acta. 2007 Jul;1774(7):856-65. doi: 10.1016/j.bbapap.2007.04.010. Epub 2007 May 5.
An enzyme that catalyzes the conversion of N-hydroxy-l-phenylalanine to phenylacetaldoxime was shown to be present in the Z-phenylacetaldoxime-degrading bacterium, Bacillus sp. strain OxB-1. The aldoxime-forming enzyme, which is induced by L-phenylalanine, was purified 8,050-fold to apparent homogeneity with a yield of 15.2%. The enzyme has a subunit M(r) of about 86,000. The enzyme converts N-hydroxy-L-phenylalanine (K(m) 0.99 mM) to only one geometrical isomer, namely Z-phenylacetaldoxime. Relatively large amounts of pyridoxal 5'-phosphate (PLP) are required to be present in the reaction mixture because PLP reacts non-enzymatically with the N-hydroxy amino acid substrate to form a nitrone. Several characteristics of the enzyme were compared with those of other PLP-dependent aromatic amino acid-converting enzymes described in the literature. The enzyme is tentatively named "N-hydroxy-L-phenylalanine decarboxylase/oxidase". Finally, the possible biosynthesis and metabolism of phenylacetaldoxime in Bacillus sp. strain OxB-1 is discussed.
一种催化N-羟基-L-苯丙氨酸转化为苯乙醛肟的酶被证明存在于降解Z-苯乙醛肟的细菌芽孢杆菌属OxB-1菌株中。由L-苯丙氨酸诱导产生的醛肟形成酶被纯化了8050倍,达到表观均一性,产率为15.2%。该酶的亚基分子量约为86,000。该酶将N-羟基-L-苯丙氨酸(Km为0.99 mM)仅转化为一种几何异构体,即Z-苯乙醛肟。反应混合物中需要存在相对大量的磷酸吡哆醛(PLP),因为PLP会与N-羟基氨基酸底物发生非酶反应形成硝酮。将该酶的几个特性与文献中描述的其他依赖PLP的芳香族氨基酸转化酶的特性进行了比较。该酶被暂定为“N-羟基-L-苯丙氨酸脱羧酶/氧化酶”。最后,讨论了芽孢杆菌属OxB-1菌株中苯乙醛肟可能的生物合成和代谢。