Engineering Polyhistidine Tags on Surface Proteins of Acidithiobacillus ferrooxidans: Impact of Localization on the Binding and Recovery of Divalent Metal Cations,ACS Applied Materials & Interfaces

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Engineering Polyhistidine Tags on Surface Proteins of Acidithiobacillus ferrooxidans: Impact of Localization on the Binding and Recovery of Divalent Metal Cations
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2022-02-16 , DOI: 10.1021/acsami.1c23682
Heejung Jung ₁ , Yuta Inaba ₁ , Virginia Jiang ₁ , Alan C West ₁ , Scott Banta ₁

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Metal processing using microorganisms has many advantages including the potential for reduced environmental impacts as compared to conventional technologies.Acidithiobacillus ferrooxidansis an iron- and sulfur-oxidizing chemolithoautotroph that is known to participate in metal bioleaching, and its metabolic capabilities have been exploited for industrial-scale copper and gold biomining. In addition to bioleaching, microorganisms could also be engineered for selective metal binding, enabling new opportunities for metal bioseparation and recovery. Here, we explored the ability of polyhistidine (polyHis) tags appended to two recombinantly expressed endogenous proteins to enhance the metal binding capacity of A. ferrooxidans. The genetically engineered cells achieved enhanced cobalt and copper binding capacities, and the Langmuir isotherm captures their interaction behavior with these divalent metals. Additionally, the cellular localization of the recombinant proteins correlated with kinetic modeling of the binding interactions, where the outer membrane-associated polyHis-tagged licanantase peptide bound the metals faster than the periplasmically expressed polyHis-tagged rusticyanin protein. The selectivity of the polyHis sequences for cobalt over copper from mixed metal solutions suggests potential utility in practical applications, and further engineering could be used to create metal-selective bioleaching microorganisms.

中文翻译：

氧化亚铁硫杆菌表面蛋白工程多组氨酸标签：定位对二价金属阳离子结合和回收的影响

与传统技术相比，使用微生物进行金属加工具有许多优势，包括减少环境影响的潜力。Acidithiobacillus ferrooxidans是一种铁和硫氧化性化学自养生物，已知参与金属生物浸出，其代谢能力已被用于工业规模的铜和金生物开采。除了生物浸出之外，还可以对微生物进行工程改造以实现选择性金属结合，从而为金属生物分离和回收提供新的机会。在这里，我们探讨了附加到两个重组表达的内源蛋白上的多组氨酸 (polyHis) 标签增强A. ferrooxidans的金属结合能力的能力. 基因工程细胞实现了增强的钴和铜结合能力，朗缪尔等温线捕捉了它们与这些二价金属的相互作用行为。此外，重组蛋白的细胞定位与结合相互作用的动力学模型相关，其中外膜相关的多组氨酸标记的葡聚糖酶肽比周质表达的多组组氨酸标记的锈菌青蛋白更快地结合金属。来自混合金属溶液的多组氨酸序列对钴对铜的选择性表明在实际应用中具有潜在的用途，并且进一步的工程可用于创建金属选择性生物浸出微生物。

更新日期：2022-02-16

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