Abstract
The copper specific binding metallothionein (CuMT) is a type of cysteine-rich, metal-binding, small protein which plays an important role in Cu2+ metabolism in vertebrates. In this study, we investigated the metal tolerance and removing ability of recombinant strains harboring CuMT obtained in vivo from the freshwater crab Sinopotamon henanense (ShCuMT) in order to study its physiological functions and metal binding capacity. We performed a 3D modeling of ShCuMT and created its structural and functional models using the I‐TASSER program. The shCumt gene was inserted into a pGEX-4t-1 vector and recombinant soluble ShCuMT was expressed in Escherichia coli. In addition, in order to characterize the tolerance and removing ability of heavy metals in E. coli with ShCuMT expression, the recombinant strains harboring ShCuMT were exposed to various concentrations of Cd2+, Cu2+ and Zn2+, respectively. The results showed that ShCuMT contains transition metal binding sites. In addition, E. coli cells expressing ShCuMT exhibited enhanced metal tolerance and higher removing ability of metal ions than control cells. However, compared with Cd2+ and Zn2+, E. coli cells expressing ShCuMT have stronger tolerance and higher removing ability of Cu2+. In general, ShCuMT contains multiple transition metal binding sites, and it could enhance tolerance and removing ability of metal ions. Therefore, ShCuMT can provide potential candidates for heavy metal bioremediation. This research on the metal binding properties of ShCuMT provides a scientific basis for bioremediation of heavy metal pollution by the recombinant strains.
Highlights
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ShCuMT recombinant strain expression enhanced Cd2+, Cu2+ and Zn2+ tolerance.
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ShCuMT recombinant strain expression enhanced Cd2+, Cu2+ and Zn2+ resistance.
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ShCuMT recombinant strain expression enhanced Cd2+, Cu2+ and Zn2+ removing ability.
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ShCuMT can be used as a regulatory biomolecule for Cu2+ homeostasis in Sinopotamon henanense.
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Acknowledgements
This work was supported by National Natural Science Foundation of China (No. 31672293), Shanxi Scholarship Council of China (No. 2016-1) and Shanxi Key Research and Development Program of China (No. 201703D221008-3). Thanks to Professors LW and CMC for their suggestions on an earlier draft of the manuscript.
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by LW, HZY, WLM, CMC and LW. The first draft of the manuscript was written by LW and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Wang, L., Yang, H.Z., Ma, W.L. et al. Study on metal binding capacity of the freshwater crab Sinopotamon henanense’s recombinant copper specific binding metallothionein expressed in Escherichia coli. Ecotoxicology 31, 149–160 (2022). https://doi.org/10.1007/s10646-021-02470-x
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DOI: https://doi.org/10.1007/s10646-021-02470-x