Clugston S L, Daub E, Kinach R, Miedema D, Barnard J F, Honek J F
Department of Chemistry, University of Waterloo, Ont., Canada.
Gene. 1997 Feb 20;186(1):103-11. doi: 10.1016/s0378-1119(96)00691-9.
The glyoxalase I gene (gloA) from Salmonella typhimurium has been isolated in Escherichia coli on a multi-copy pBR322-derived plasmid, selecting for resistance to 3 mM methylglyoxal on Luria-Bertani agar. The region of the plasmid which confers the methylglyoxal resistance in E. coli was sequenced. The deduced protein sequence was compared to the known sequences of the Homo sapiens and Pseudomonas putida glyoxalase I (GlxI) enzymes, and regions of strong homology were used to probe the National Center for Biotechnology Information protein database. This search identified several previously known glyoxalase I sequences and other open reading frames with unassigned function. The clustal alignments of the sequences are presented, indicating possible Zn2+ ligands and active site regions. In addition, the S. typhimurium sequence aligns with both the N-terminal half and the C-terminal half of the proposed GlxI sequences from Saccharomyces cerevisiae and Schizosaccharomyces pombe, suggesting that the structures of the yeast enzymes are those of fused dimers.
鼠伤寒沙门氏菌的乙二醛酶I基因（gloA）已在大肠杆菌中通过多拷贝pBR322衍生质粒分离出来，通过在Luria-Bertani琼脂上筛选对3 mM甲基乙二醛的抗性来实现。对在大肠杆菌中赋予甲基乙二醛抗性的质粒区域进行了测序。将推导的蛋白质序列与智人和恶臭假单胞菌乙二醛酶I（GlxI）酶的已知序列进行比较，并使用高度同源区域对美国国立生物技术信息中心的蛋白质数据库进行搜索。该搜索鉴定出了几个先前已知的乙二醛酶I序列以及其他功能未明确的开放阅读框。给出了序列的Clustal比对结果，显示了可能的Zn2+配体和活性位点区域。此外，鼠伤寒沙门氏菌序列与酿酒酵母和粟酒裂殖酵母中推测的GlxI序列的N端一半和C端一半均比对一致，这表明酵母酶的结构是融合二聚体的结构。
