Tandem oligomeric expression of metallothionein enhance heavy metal tolerance and bioaccumulation in Escherichia coli.

2.50
Hdl Handle:
http://hdl.handle.net/10547/623349
Title:
Tandem oligomeric expression of metallothionein enhance heavy metal tolerance and bioaccumulation in Escherichia coli.
Authors:
Ma, Wenli; Li, Xuefen; Wang, Qi; Ren, Zhumei; Crabbe, M. James C. ( 0000-0003-3609-1963 ) ; Wang, Lan
Abstract:
Metallothioneins (MTs) are a family of low molecular weight, cysteine-rich, metal-binding proteins, which play important roles in metal homeostasis and heavy metal detoxification. In our previous study, a novel full length MT cDNA was successfully cloned from the freshwater crab (Sinopotamon henanense). In the present study, tandem repeats of two and three copies of the crab MT gene were integrated by overlap extension PCR (SOEPCR) and expressed in Escherichia coli. The SUMO fusion expression system was adopted to increase the stability and solubility of the recombinant MT proteins. The recombinant proteins were purified and their metal-binding abilities were further analyzed by the ultraviolet absorption spectral scan. Furthermore, the metal tolerance and bioaccumulation of E. coli cells expressing oligomeric MTs were determined. Results showed that the recombinant plasmids pET28a-SUMO-2MT and pET28a-SUMO-3MT were successfully constructed. SDS-PAGE analysis showed that the SUMO-2MT and SUMO-3MT were expressed mainly in the soluble forms. Oligomeric MTs expression significantly enhanced Cu, Cd or Zn tolerance and accumulation in E. coli in the order: SUMO-3MT˃SUMO-2MT˃SUMO-MT˃control. Cells harboring pET28a-SUMO -3MT exhibited the highest Cu, Cd or Zn bioaccumulation at 5.8-fold, 3.1-fold or 6.7-fold higher than that of the control cells. Our research could lay a foundation for large-scale preparation of MTs and provide a scientific basis for bioremediation of heavy metal pollution by oligomeric MTs.
Affiliation:
Shanxi University; University of Oxford; University of Bedfordshire
Citation:
Ma W, Li X, Wang Q, Ren Z, Crabbe MJC, Wang L (2019) 'Tandem oligomeric expression of metallothionein enhance heavy metal tolerance and bioaccumulation in Escherichia coli.', Ecotoxicology and Environmental Safety, 181 (), pp.301-307.
Publisher:
Elsevier
Journal:
Ecotoxicology and Environmental Safety
Issue Date:
13-Jun-2019
URI:
http://hdl.handle.net/10547/623349
DOI:
10.1016/j.ecoenv.2019.06.022
Additional Links:
https://www.sciencedirect.com/science/article/pii/S0147651319306608
Type:
Article
Language:
en
ISSN:
0147-6513
EISSN:
0147-6513
Appears in Collections:
Biomedical and biological science

Full metadata record

DC FieldValue Language
dc.contributor.authorMa, Wenlien
dc.contributor.authorLi, Xuefenen
dc.contributor.authorWang, Qien
dc.contributor.authorRen, Zhumeien
dc.contributor.authorCrabbe, M. James C.en
dc.contributor.authorWang, Lanen
dc.date.accessioned2019-07-08T10:03:35Z-
dc.date.available2019-07-08T10:03:35Z-
dc.date.issued2019-06-13-
dc.identifier.citationMa W, Li X, Wang Q, Ren Z, Crabbe MJC, Wang L (2019) 'Tandem oligomeric expression of metallothionein enhance heavy metal tolerance and bioaccumulation in Escherichia coli.', Ecotoxicology and Environmental Safety, 181 (), pp.301-307.en
dc.identifier.issn0147-6513-
dc.identifier.doi10.1016/j.ecoenv.2019.06.022-
dc.identifier.urihttp://hdl.handle.net/10547/623349-
dc.description.abstractMetallothioneins (MTs) are a family of low molecular weight, cysteine-rich, metal-binding proteins, which play important roles in metal homeostasis and heavy metal detoxification. In our previous study, a novel full length MT cDNA was successfully cloned from the freshwater crab (Sinopotamon henanense). In the present study, tandem repeats of two and three copies of the crab MT gene were integrated by overlap extension PCR (SOEPCR) and expressed in Escherichia coli. The SUMO fusion expression system was adopted to increase the stability and solubility of the recombinant MT proteins. The recombinant proteins were purified and their metal-binding abilities were further analyzed by the ultraviolet absorption spectral scan. Furthermore, the metal tolerance and bioaccumulation of E. coli cells expressing oligomeric MTs were determined. Results showed that the recombinant plasmids pET28a-SUMO-2MT and pET28a-SUMO-3MT were successfully constructed. SDS-PAGE analysis showed that the SUMO-2MT and SUMO-3MT were expressed mainly in the soluble forms. Oligomeric MTs expression significantly enhanced Cu, Cd or Zn tolerance and accumulation in E. coli in the order: SUMO-3MT˃SUMO-2MT˃SUMO-MT˃control. Cells harboring pET28a-SUMO -3MT exhibited the highest Cu, Cd or Zn bioaccumulation at 5.8-fold, 3.1-fold or 6.7-fold higher than that of the control cells. Our research could lay a foundation for large-scale preparation of MTs and provide a scientific basis for bioremediation of heavy metal pollution by oligomeric MTs.en
dc.language.isoenen
dc.publisherElsevieren
dc.relation.urlhttps://www.sciencedirect.com/science/article/pii/S0147651319306608en
dc.rightsGreen - can archive pre-print and post-print or publisher's version/PDF-
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectmetal toleranceen
dc.subjectC150 Environmental Biologyen
dc.titleTandem oligomeric expression of metallothionein enhance heavy metal tolerance and bioaccumulation in Escherichia coli.en
dc.typeArticleen
dc.identifier.eissn0147-6513-
dc.contributor.departmentShanxi Universityen
dc.contributor.departmentUniversity of Oxforden
dc.contributor.departmentUniversity of Bedfordshireen
dc.identifier.journalEcotoxicology and Environmental Safetyen
dc.date.updated2019-07-08T10:00:25Z-
dc.description.noteUnfortunately Elsevier will not allow us to archive the final published version, do you have a previous version we could use? E.g. after review but before publisher formatting applied? supplied.-
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