Lal Piyush Behari, Schneider Barbara L, Vu Khoan, Reitzer Larry
Department of Molecular and Cell Biology, The University of Texas at Dallas, Richardson, TX, 75080, USA.
Mol Microbiol. 2014 Nov;94(4):843-56. doi: 10.1111/mmi.12801. Epub 2014 Oct 12.
Aminotransferases can be redundant or promiscuous, but the extent and significance of these properties is not known in any organism, even in Escherichia coli. To determine the extent of redundancy, it was first necessary to identify the redundant aminotransferases in arginine and lysine synthesis, and then complement all aminotransferase-deficient mutants with genes for all aminotransferases. The enzymes with N-acetylornithine aminotransferase (ACOAT) activity in arginine synthesis were ArgD, AstC, GabT and PuuE; the major anaerobic ACOAT was ArgD. The major enzymes with N-succinyl-l,l-diaminopimelate aminotransferase (SDAP-AT) activity in lysine synthesis were ArgD, AstC, and SerC. Seven other aminotransferases, when overproduced, complemented the defect in a triple mutant. Lysine availability did not regulate synthesis of the major SDAP-ATs. Complementation analysis of mutants lacking aminotransferases showed that the SDAP-ATs and alanine aminotransferases were exceptionally redundant, and it is proposed that this redundancy may ensure peptidoglycan synthesis. An overview of all aminotransferase reactions indicates that redundancy and broad specificity are common properties of aminotransferases.
氨基转移酶可能存在冗余或具有多底物适应性,但在任何生物体中,甚至在大肠杆菌中,这些特性的程度和意义都尚不清楚。为了确定冗余程度,首先有必要在精氨酸和赖氨酸合成过程中鉴定出冗余的氨基转移酶,然后用所有氨基转移酶的基因对所有氨基转移酶缺陷型突变体进行互补。在精氨酸合成过程中具有N - 乙酰鸟氨酸氨基转移酶(ACOAT)活性的酶有ArgD、AstC、GabT和PuuE;主要的厌氧ACOAT是ArgD。在赖氨酸合成过程中具有N - 琥珀酰 - L,L - 二氨基庚二酸氨基转移酶(SDAP - AT)活性的主要酶是ArgD、AstC和SerC。另外七种氨基转移酶在过量表达时,能够互补三重突变体中的缺陷。赖氨酸的可利用性并不调节主要SDAP - AT的合成。对缺乏氨基转移酶的突变体进行互补分析表明,SDAP - AT和丙氨酸氨基转移酶具有极高的冗余性,有人提出这种冗余性可能确保肽聚糖的合成。对所有氨基转移酶反应的概述表明,冗余和广泛特异性是氨基转移酶的共同特性。