Barrette W C, Hannum D M, Wheeler W D, Hurst J K
Department of Chemical and Biological Sciences, Oregon Graduate Center, Beaverton 97006-1999.
J Bacteriol. 1988 Aug;170(8):3655-9. doi: 10.1128/jb.170.8.3655-3659.1988.
Metabolic regulation by nucleotides has been examined in several bacteria within the context of the adenylate energy charge (EC) concept. The ECs of bacteria capable of only fermentative metabolism (Streptococcus lactis and the ATPase-less mutant Escherichia coli AN718) fell to less than 0.2 under carbon-limiting conditions, but the bacteria were able to step up the EC to greater than 0.8 upon exposure to nutrient sugars. Similarly, nongrowing E. coli 25922, whose EC had been artificially lowered to less than 0.1 by the addition of the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), was able to immediately step up the EC to 0.8 to 0.9 upon the addition of glucose but was unable to respond to respiratory substrates. The EC of respiring bacteria (E. coli 25922 and Pseudomonas aeruginosa 27853) fell to 0.3 to 0.4 under certain limiting growth conditions, but the bacteria also responded immediately when challenged with succinate to give EC values greater than 0.8. These bacteria could not step up the EC with respiratory substrates in the presence of CCCP. For all bacteria, the loss of the ability to step up the EC was attributable to the loss of nutrient transport function. Mixtures of viable and HOCl-killed E. coli 25922 were able to step up the EC in proportion to the fraction of surviving cells. The data indicate that nucleotide phosphorylation levels are not regulatory in nongrowing bacteria but that the EC step-up achievable upon nutrient addition may be an accurate index of viability.
在腺苷酸能荷(EC)概念的背景下,已经在几种细菌中研究了核苷酸的代谢调节。仅具有发酵代谢能力的细菌(乳酸链球菌和无ATP酶的大肠杆菌突变体AN718)在碳限制条件下,其能荷降至0.2以下,但在接触营养糖后,这些细菌能够将能荷提高到0.8以上。同样,未生长的大肠杆菌25922,其能荷通过添加质子载体间氯苯腙(CCCP)被人为降低至0.1以下,在添加葡萄糖后能够立即将能荷提高到0.8至0.9,但对呼吸底物无反应。进行呼吸作用的细菌(大肠杆菌25922和铜绿假单胞菌27853)在某些限制生长条件下,其能荷降至0.3至0.4,但当用琥珀酸刺激时,这些细菌也能立即做出反应,使能荷值大于0.8。在存在CCCP的情况下,这些细菌不能通过呼吸底物提高能荷。对于所有细菌,提高能荷的能力丧失归因于营养物质转运功能的丧失。活的和经次氯酸杀死的大肠杆菌25922的混合物能够根据存活细胞的比例提高能荷。数据表明,在未生长的细菌中,核苷酸磷酸化水平不具有调节作用,但添加营养物质后可实现的能荷提高可能是活力的准确指标。