首页> 美国卫生研究院文献>Applied and Environmental Microbiology >The methanogenic archaeon Methanosarcina thermophila TM-1 possesses a high-affinity glycine betaine transporter involved in osmotic adaptation.
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The methanogenic archaeon Methanosarcina thermophila TM-1 possesses a high-affinity glycine betaine transporter involved in osmotic adaptation.

机译:产甲烷的古生甲烷菌(Methanosarcina thermophila)TM-1具有参与渗透性适应的高亲和力甘氨酸甜菜碱转运蛋白。

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摘要

Methanogenic Archaea are found in a wide range of environments and use several strategies to adjust to changes in extracellular solute concentrations. One methanogenic archaeon, Methanosarcina thermophila TM-1, can adapt to various osmotic conditions by synthesis of alpha-glutamate and a newly discovered compatible solute, Ne-acetyl-beta-lysine, or by accumulation of glycine betaine (betaine) and potassium ions from the environment. Since betaine transport has not been characterized for any of the methanogenic Archaea, we examined the uptake of this solute by M. thermophila TM-1. When cells were grown in mineral salts media containing from 0.1 to 0.8 M NaC1, M. thermophila accumulated betaine in concentrations up to 140 times those of a concentration gradient within 10 min of exposure to the solute. The betaine uptake system consisted of a single, high-affinity transporter with an apparent K3 of 10 microM and an apparent maximum transport velocity of 1.15 nmol/min/mg of protein. The transporter appeared to be specific for betaine, since potential substrates, including glycine, sarcosine, dimethyl glycine, choline, and proline, did not significantly inhibit betaine uptake. M. thermophila TM-1 cells can also regulate the capacity for betaine accumulation, since the rate of betaine transport was reduced in cells pregrown in a high-osmolarity medium when 500 microM betaine was present. Betaine transport appears to be H+ and/or Na+ driven, since betaine transport was inhibited by several types of protonophores and sodium ionophores.
机译:产甲烷古生菌存在于广泛的环境中,并使用多种策略来适应细胞外溶质浓度的变化。一种产甲烷的古菌甲烷嗜热甲烷菌(Methanosarcina thermophila TM-1)可以通过合成α-谷氨酸和新发现的相容性溶质Ne-乙酰-β-赖氨酸,或通过积累甘氨酸甜菜碱(甜菜碱)和钾离子来适应各种渗透条件。环境。由于尚无任何产甲烷甲烷菌的甜菜碱转运特征,因此我们研究了嗜热毁丝霉TM-1对这种溶质的吸收。当细胞在含有0.1至0.8 M NaCl的矿物盐培养基中生长时,嗜热毁丝霉会在暴露于溶质的10分钟内累积甜菜碱,其浓度高达浓度梯度的140倍。甜菜碱摄取系统由单个高亲和力转运蛋白组成,其表观K3为10 microM,表观最大转运速度为1.15 nmol / min / mg蛋白质。转运蛋白似乎对甜菜碱具有特异性,因为潜在的底物(包括甘氨酸,肌氨酸,二甲基甘氨酸,胆碱和脯氨酸)不会显着抑制甜菜碱的摄取。嗜热毁丝霉TM-1细胞还可以调节甜菜碱的积累能力,因为当存在500 microM甜菜碱时,高渗透压培养基中预生长的细胞中甜菜碱的转运速率会降低。甜菜碱的运输似乎是由H +和/或Na +驱动的,因为甜菜碱的运输受到几种类型的质子载体和钠离子载体的抑制。

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