对羟苯甲酸3-羟化酶初步纯化及进化起源探讨

Preliminary Purification and Pursuit of Evolutionary Origin of P-hydroxybenzoate 3-hydroxylase from Comamonas Testosteroni

经超声破碎、硫酸铵分级沉淀、凝胶过滤、离子交换层析和SDS PAGE ,从Comamonastestosteroni菌株中获得了相对分子质量为 4 .5万的对羟苯甲酸 3 羟化酶 (POB3 HOase) ;测定了其N 末端 2 1个氨基酸序列 ,并与文献报道来源于不同菌株的POB3 HOase序列进行了同源性比较 ,结果提示它们具有相同的进化起源。

The P-hydroxybenzoate 3-hydroxylase (POB-HOase) with 45 thousand relative molecular mass from Comamonas testosteroni was purified by using sonic crushing, ammonium sulfate fractionation, molecular sieve chromatography, exchange chromatography and SDS-PAGE. The N-termina lamino acid sequence was determined up to 21 residues. The amino acid sequence was compared with the known N-terminal sequences of POB3-HOases from Pseudomonas aeruginosa, Pseudomonas fluorescens and Acinetobacter calcoaceticus. The high degree of homology observed between the POB3-HOases from Comamonas and those of Pseudomonas and Acinetobacter indicates the common evolutionary origin of POB3-HOase among these soil bacteria of different genera.