Peck M W
Institute of Food Research, Norwich Laboratory, United Kingdom.
Appl Environ Microbiol. 1989 Apr;55(4):940-5. doi: 10.1128/aem.55.4.940-945.1989.
Coenzyme F420 has been assayed by high-performance liquid chromatography with fluorimetric detection; this permits quantification of individual coenzyme F420 analogs whilst avoiding the inclusion of interfering material. The total intracellular coenzyme F420 content of Methanosarcina barkeri MS cultivated on methanol and on H2-CO2 and of Methanosarcina mazei S-6 cultured on methanol remained relatively constant during batch growth. The most abundant analogs in M. barkeri were coenzymes F420-2 and F420-4, whilst in M. mazei coenzymes F420-2 and F420-3 predominated. Significant changes in the relative proportions of the coenzyme F420 analogs were noted during batch growth, with coenzymes F420-2 and F420-4 showing opposite responses to each other and the same being also true for coenzymes F420-3 and F420-5. This suggests that an enzyme responsible for transferring pairs of glutamic acid residues may be active. The degradation fragment FO was also detected in cells in late exponential and stationary phase. Coenzyme F420 analogs were present in the culture supernatant of both methanogens, in similar proportions to that in the cells, except for FO which was principally located in the supernatant.
辅酶F420已通过高效液相色谱荧光检测法进行测定;这使得能够对单个辅酶F420类似物进行定量,同时避免包含干扰物质。在甲醇和H2-CO2上培养的巴氏甲烷八叠球菌MS以及在甲醇上培养的马氏甲烷八叠球菌S-6的细胞内辅酶F420总含量在分批培养过程中保持相对恒定。巴氏甲烷八叠球菌中最丰富的类似物是辅酶F420-2和F420-4,而在马氏甲烷八叠球菌中辅酶F420-2和F420-3占主导。在分批培养过程中,注意到辅酶F420类似物的相对比例有显著变化,辅酶F420-2和F420-4呈现相反的响应,辅酶F420-3和F420-5也是如此。这表明负责转移谷氨酸残基对的一种酶可能是有活性的。在指数后期和稳定期的细胞中也检测到了降解片段FO。两种产甲烷菌的培养上清液中都存在辅酶F420类似物,其比例与细胞中的相似,但FO主要位于上清液中。