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代谢多功能细菌沼泽红假单胞菌 TIE-1 中铁和锰运输中的遗传冗余。

Genetic Redundancy in Iron and Manganese Transport in the Metabolically Versatile Bacterium Rhodopseudomonas palustris TIE-1.

机构信息

Department of Biology, Washington University in St. Louis, St. Louis, Missouri, USA.

Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis, Missouri, USA.

出版信息

Appl Environ Microbiol. 2020 Aug 3;86(16). doi: 10.1128/AEM.01057-20.

Abstract

The purple nonsulfur bacterium TIE-1 can produce useful biochemicals such as bioplastics and biobutanol. Production of such biochemicals requires intracellular electron availability, which is governed by the availability and the transport of essential metals such as iron (Fe). Because of the distinct chemical properties of ferrous [Fe(II)] and ferric iron [Fe(III)], different systems are required for their transport and storage in bacteria. Although Fe(III) transport systems are well characterized, we know much less about Fe(II) transport systems except for the FeoAB system. Iron transporters can also import manganese (Mn). We studied Fe and Mn transport by five putative Fe transporters in TIE-1 under metal-replete, metal-depleted, oxic, and anoxic conditions. We observed that by overexpressing , , and , the intracellular concentrations of Fe and Mn can be enhanced in TIE-1 under oxic and anoxic conditions, respectively. The deletion of a single gene/operon does not attenuate Fe or Mn uptake in TIE-1 regardless of the growth conditions used. This indicates that genetically dissimilar yet functionally redundant Fe transporters in TIE-1 can complement each other. Relative gene expression analysis shows that and are expressed during Fe and Mn depletion under both oxic and anoxic conditions. The promoters of these transporter genes contain a combination of Fur and Fnr boxes, suggesting that their expression is regulated by both Fe and oxygen availability. The findings from this study will help us modulate intracellular Fe and Mn concentrations, ultimately improving TIE-1's ability to produce desirable biomolecules. TIE-1 is a metabolically versatile bacterium that can use various electron donors, including Fe(II) and poised electrodes, for photoautotrophic growth. TIE-1 can produce useful biomolecules, such as biofuels and bioplastics, under various growth conditions. Production of such reduced biomolecules is controlled by intracellular electron availability, which, in turn, is mediated by various iron-containing proteins in the cell. Several putative Fe transporters exist in TIE-1's genome. Some of these transporters can also transport Mn, part of several important cellular enzymes. Therefore, understanding the ability to transport and respond to various levels of Fe and Mn under different conditions is important to improve TIE-1's ability to produce useful biomolecules. Our data suggest that by overexpressing Fe transporter genes via plasmid-based expression, we can increase the import of Fe and Mn in TIE-1. Future work will leverage these data to improve TIE-1 as an attractive microbial chassis and future biotechnological workhorse.

摘要

紫色非硫细菌 TIE-1 可以生产有用的生物化学物质,如生物塑料和生物丁醇。这些生物化学物质的生产需要细胞内电子的可用性,这由必需金属(如铁 (Fe))的可用性和运输来控制。由于二价铁 [Fe(II)] 和三价铁 [Fe(III)] 的化学性质明显不同,因此需要不同的系统来运输和储存它们。尽管 Fe(III) 转运系统已经得到很好的描述,但除了 FeoAB 系统之外,我们对 Fe(II) 转运系统的了解要少得多。铁转运蛋白还可以导入锰 (Mn)。在金属充足、金属耗尽、需氧和缺氧条件下,我们研究了 TIE-1 中五个假定的 Fe 转运蛋白对 Fe 和 Mn 的运输。我们观察到,过表达 、 和 可以分别增强 TIE-1 在需氧和缺氧条件下的细胞内 Fe 和 Mn 浓度。无论使用何种生长条件,单个基因/操纵子的缺失都不会削弱 TIE-1 对 Fe 或 Mn 的摄取。这表明 TIE-1 中遗传上不同但功能上冗余的 Fe 转运蛋白可以相互补充。相对基因表达分析表明,在需氧和缺氧条件下,当 Fe 和 Mn 耗尽时, 和 都有表达。这些转运蛋白基因的启动子包含 Fur 和 Fnr 框的组合,表明它们的表达受到 Fe 和氧气可用性的共同调节。这项研究的结果将帮助我们调节细胞内 Fe 和 Mn 的浓度,最终提高 TIE-1 生产理想生物分子的能力。TIE-1 是一种代谢多功能细菌,可使用各种电子供体,包括 Fe(II) 和 poised 电极,进行光自养生长。TIE-1 可以在各种生长条件下生产有用的生物分子,如生物燃料和生物塑料。这些还原生物分子的产生受细胞内电子可用性的控制,而电子可用性又受细胞内各种含铁蛋白的调节。TIE-1 基因组中存在几个假定的 Fe 转运蛋白。其中一些转运蛋白还可以运输 Mn,这是几种重要细胞酶的一部分。因此,了解在不同条件下运输和响应各种水平的 Fe 和 Mn 的能力对于提高 TIE-1 生产有用生物分子的能力非常重要。我们的数据表明,通过质粒表达过表达 Fe 转运蛋白基因,我们可以增加 TIE-1 对 Fe 和 Mn 的摄取。未来的工作将利用这些数据来提高 TIE-1 作为有吸引力的微生物底盘和未来生物技术骨干的能力。

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