Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario K1A 0R6, Canada.
Appl Environ Microbiol. 1993 Apr;59(4):1092-8. doi: 10.1128/aem.59.4.1092-1098.1993.
Eleven strains of methanogenic bacteria were divided into two groups on the basis of the directionality (oxidative or reductive) of their citric acid pathways. These pathways were readily identified for most methanogens from the patterns of carbon atom labeling in glutamate, following growth in the presence of [2-C]acetate. All used noncyclic pathways, but members of the family Methanosarcinaceae were the only methanogens found to use the oxidative direction. Methanococcus jannaschii failed to incorporate carbon from acetate despite transmembrane equilibration comparable to other weak acids. This organism was devoid of detectable activities of the acetate-incorporating enzymes acetyl coenzyme A synthetase, acetate kinase, and phosphotransacetylase. However, incorporation of [1-C]-, [2-C]-, or [3-C]pyruvate during the growth of M. jannaschii was possible and resulted in labeling patterns indicative of a noncyclic citric acid pathway operating in the reductive direction to synthesize amino acids. Carbohydrates were labeled consistent with glucogenesis from pyruvate. Leucine, isoleucine, phenylalanine, lysine, formate, glycerol, and mevalonate were incorporated when supplied to the growth medium. Lysine was preferentially incorporated into the lipid fraction, suggesting a role as a phytanyl chain precursor.
根据柠檬酸途径的方向性(氧化或还原),将 11 株产甲烷菌分为两组。通过在[2-C]乙酸盐存在下生长时谷氨酸中碳原子标记模式,可以很容易地识别大多数产甲烷菌的这些途径。所有产甲烷菌都使用非循环途径,但仅在 Methanosarcinaceae 科中发现产甲烷菌使用氧化方向。尽管跨膜平衡与其他弱酸相当,但 Methanococcus jannaschii 未能从乙酸盐中掺入碳。该生物缺乏可检测到的乙酰辅酶 A 合成酶、乙酸激酶和磷酸转乙酰酶等掺入乙酸盐的酶的活性。然而,在 M. jannaschii 的生长过程中,[1-C]、[2-C]或[3-C]丙酮酸的掺入是可能的,并导致表明非循环柠檬酸途径在还原方向起作用以合成氨基酸的标记模式。碳水化合物的标记与来自丙酮酸的糖异生一致。当供应到生长培养基中时,掺入了亮氨酸、异亮氨酸、苯丙氨酸、赖氨酸、甲酸盐、甘油和甲羟戊酸。赖氨酸优先掺入脂质部分,表明其作为植烷链前体的作用。