Wakayama M, Ashika T, Miyamoto Y, Yoshikawa T, Sonoda Y, Sakai K, Moriguchi M
Department of Applied Chemistry, Faculty of Engineering, Oita University.
J Biochem. 1995 Jul;118(1):204-9. doi: 10.1093/oxfordjournals.jbchem.a124879.
The gene coding the N-acyl-D-glutamate amidohydrolase of Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (Alcaligenes A-6) was cloned and its complete DNA sequence was determined. The N-acyl-D-glutamate amidohydrolase structural gene consists of 1,464 nucleotides and encodes 488 amino acid residues. The molecular weight of the enzyme was calculated to be 51,490. This value is close to the apparent molecular weight of 59,000 determined for the purified enzyme from Alcaligenes A-6 by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). The N-terminal amino acid sequence of the recombinant protein exactly matches the amino acid sequence derived from the DNA sequence and that determined from the Alcaligenes A-6 enzyme (NH2-MQEKLDLVIEGGWVIDGLGG). The deduced amino acid sequence of the cloned N-acyl-D-glutamate amidohydrolase showed high sequence homology with those of N-acyl-D-aspartate amidohydrolase (46%) and D-aminoacylase (47%) from Alcaligenes A-6. This fact strongly suggests that these three enzymes have evolved from a common ancestral gene.
对木糖氧化产碱杆菌木糖氧化亚种A-6（产碱杆菌A-6）的N-酰基-D-谷氨酸酰胺水解酶编码基因进行了克隆，并测定了其完整的DNA序列。N-酰基-D-谷氨酸酰胺水解酶结构基因由1464个核苷酸组成，编码488个氨基酸残基。计算该酶的分子量为51490。该值与通过十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳（PAGE）测定的产碱杆菌A-6纯化酶的表观分子量59000接近。重组蛋白的N端氨基酸序列与从DNA序列推导的氨基酸序列以及从产碱杆菌A-6酶测定的氨基酸序列（NH2-MQEKLDLVIEGGWVIDGLGG）完全匹配。克隆的N-酰基-D-谷氨酸酰胺水解酶推导的氨基酸序列与产碱杆菌A-6的N-酰基-D-天冬氨酸酰胺水解酶（46%）和D-氨基酰化酶（47%）的氨基酸序列具有高度的序列同源性。这一事实有力地表明这三种酶是由一个共同的祖先基因进化而来的。
