Department of Chemistry & Biochemistry, University of California, Santa Barbara, CA, United States.
Department of Chemistry & Biochemistry, University of California, Santa Barbara, CA, United States.
Methods Enzymol. 2024;702:189-214. doi: 10.1016/bs.mie.2024.06.006. Epub 2024 Jul 20.
The C-diazeniumdiolate (N-nitrosohydroxylamine) group in the amino acid graminine (Gra) is a newly discovered Fe(III) ligand in microbial siderophores. Graminine was first identified in the siderophore gramibactin, and since this discovery, other Gra-containing siderophores have been identified, including megapolibactins, plantaribactin, gladiobactin, trinickiabactin (gramibactin B), and tistrellabactins. The C-diazeniumdiolate is photoreactive in UV light which provides a convenient characterization tool for this type of siderophore. This report details the process of genomics-driven identification of bacteria producing Gra-containing siderophores based on selected biosynthetic enzymes, as well as bacterial culturing, isolation and characterization of the C-diazeniumdiolate siderophores containing Gra.
氨基酸谷氨酰胺(Gra)中的 C-二氮烯二酸盐(亚硝基羟胺)基团是微生物铁载体中一种新发现的 Fe(III)配体。谷氨酰胺首先在铁载体gramibactin 中被鉴定出来,自这一发现以来,其他含有 Gra 的铁载体也被鉴定出来,包括megapolibactins、plantaribactin、gladiobactin、trinickiabactin(gramibactin B)和 tistrellabactins。C-二氮烯二酸盐在紫外光下具有光反应性,这为这种类型的铁载体提供了一个方便的特征描述工具。本报告详细介绍了基于选定生物合成酶的基因组学驱动的生产含有 Gra 的铁载体的细菌的鉴定过程,以及含有 Gra 的 C-二氮烯二酸盐铁载体的细菌培养、分离和特性描述。