Pflüger K, Baumann S, Gottschalk G, Lin W, Santos H, Müller V
Section of Microbiology, Department of Biology I, Ludwig-Maximilians-Universität München, 80638 Munich, Germany.
Appl Environ Microbiol. 2003 Oct;69(10):6047-55. doi: 10.1128/AEM.69.10.6047-6055.2003.
The compatible solute N(epsilon)-acetyl-beta-lysine is unique to methanogenic archaea and is produced under salt stress only. However, the molecular basis for the salt-dependent regulation of N(epsilon)-acetyl-beta-lysine formation is unknown. Genes potentially encoding lysine-2,3-aminomutase (ablA) and beta-lysine acetyltransferase (ablB), which are assumed to catalyze N(epsilon)-acetyl-beta-lysine formation from alpha-lysine, were identified on the chromosomes of the methanogenic archaea Methanosarcina mazei Gö1, Methanosarcina acetivorans, Methanosarcina barkeri, Methanococcus jannaschii, and Methanococcus maripaludis. The order of the two genes was identical in the five organisms, and the deduced proteins were very similar, indicating a high degree of conservation of structure and function. Northern blot analysis revealed that the two genes are organized in an operon (termed the abl operon) in M. mazei Gö1. Expression of the abl operon was strictly salt dependent. The abl operon was deleted in the genetically tractable M. maripaludis. Delta(abl) mutants of M. maripaludis no longer produced N(epsilon)-acetyl-beta-lysine and were incapable of growth at high salt concentrations, indicating that the abl operon is essential for N(epsilon)-acetyl-beta-lysine synthesis. These experiments revealed the first genes involved in the biosynthesis of compatible solutes in methanogens.
相容性溶质N(ε)-乙酰-β-赖氨酸是产甲烷古菌所特有的,且仅在盐胁迫下产生。然而,N(ε)-乙酰-β-赖氨酸形成的盐依赖性调控的分子基础尚不清楚。在产甲烷古菌马氏甲烷八叠球菌Gö1、嗜乙酸甲烷八叠球菌、巴氏甲烷八叠球菌、詹氏甲烷球菌和沼泽甲烷球菌的染色体上,鉴定出了可能编码赖氨酸-2,3-氨基变位酶(ablA)和β-赖氨酸乙酰转移酶(ablB)的基因,这两种酶被认为可催化由α-赖氨酸形成N(ε)-乙酰-β-赖氨酸。这两个基因在这五种生物中的排列顺序相同,推导的蛋白质也非常相似,表明其结构和功能具有高度保守性。Northern印迹分析显示,这两个基因在马氏甲烷八叠球菌Gö1中组成一个操纵子(称为abl操纵子)。abl操纵子的表达严格依赖于盐。在遗传上易于操作的沼泽甲烷球菌中删除了abl操纵子。沼泽甲烷球菌的Δ(abl)突变体不再产生N(ε)-乙酰-β-赖氨酸,并且在高盐浓度下无法生长,这表明abl操纵子对于N(ε)-乙酰-β-赖氨酸的合成至关重要。这些实验揭示了产甲烷菌中首个参与相容性溶质生物合成的基因。