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在大肠杆菌中异源表达的金属结合蛋白对 Hg/Cr 的吸附。

Adsorption of Hg/Cr by metal-binding proteins heterologously expressed in Escherichia coli.

机构信息

Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China.

Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, PR China.

出版信息

BMC Biotechnol. 2024 Mar 23;24(1):15. doi: 10.1186/s12896-024-00842-9.

Abstract

BACKGROUND

Removal of heavy metals from water and soil is a pressing challenge in environmental engineering, and biosorption by microorganisms is considered as one of the most cost-effective methods. In this study, the metal-binding proteins MerR and ChrB derived from Cupriavidus metallidurans were separately expressed in Escherichia coli BL21 to construct adsorption strains. To improve the adsorption performance, surface display and codon optimization were carried out.

RESULTS

In this study, we constructed 24 adsorption engineering strains for Hg and Cr, utilizing different strategies. Among these engineering strains, the M'-002 and B-008 had the strongest heavy metal ion absorption ability. The M'-002 used the flexible linker and INPN to display the merR at the surface of the E. coli BL21, whose maximal adsorption capacity reached 658.40 μmol/g cell dry weight under concentrations of 300 μM Hg. And the B-008 overexpressed the chrB in the intracellular, its maximal capacity was 46.84 μmol/g cell dry weight under concentrations 500 μM Cr. While in the case of mixed ions solution (including Pb, Cd, Cr and Hg), the total amount of ions adsorbed by M'-002 and B-008 showed an increase of up to 1.14- and 4.09-folds, compared to the capacities in the single ion solution.

CONCLUSION

The construction and optimization of heavy metal adsorption strains were carried out in this work. A comparison of the adsorption behavior between single bacteria and mixed bacteria systems was investigated in both a single ion and a mixed ion environment. The Hg absorption capacity is reached the highest reported to date with the engineered strain M'-002, which displayed the merR at the surface of chassis cell, indicating the strain's potential for its application in practical environments.

摘要

背景

从水和土壤中去除重金属是环境工程面临的紧迫挑战,而微生物的生物吸附被认为是最具成本效益的方法之一。在这项研究中,分别从铜绿假单胞菌中表达了金属结合蛋白 MerR 和 ChrB,以构建吸附菌株。为了提高吸附性能,进行了表面展示和密码子优化。

结果

本研究利用不同策略构建了 24 株用于 Hg 和 Cr 的吸附工程菌。在这些工程菌中,M'-002 和 B-008 对重金属离子的吸附能力最强。M'-002 利用柔性接头和 INPN 将 merR 展示在大肠杆菌 BL21 的表面,在 300 μM Hg 浓度下,其最大吸附量达到 658.40 μmol/g 细胞干重。而 B-008 在细胞内过表达 chrB,在 500 μM Cr 浓度下,其最大容量为 46.84 μmol/g 细胞干重。而在混合离子溶液(包括 Pb、Cd、Cr 和 Hg)中,M'-002 和 B-008 吸附的总离子量分别比单一离子溶液中的容量增加了 1.14 倍和 4.09 倍。

结论

本工作对重金属吸附菌株进行了构建和优化,并在单一离子和混合离子环境中研究了单菌和混合菌系统的吸附行为。用工程菌株 M'-002 吸附 Hg 的能力达到了迄今为止的最高水平,该菌株在底盘细胞表面展示了 merR,表明该菌株在实际环境中的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4afd/10960487/a50d327403d2/12896_2024_842_Fig1_HTML.jpg

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