Genes for dimeric and tetrameric human metallothionein (hMT) were designed and successfully overexpressed in Escherichia coli to generate functional oligomeric hMTs. An hMT synthesized with prokaryotic codons, a linker encoding a gly-gly-gly tripeptide, and Met-deficient hMT-II was ligated to create a dimeric hMT, from which a tetrameric hMT was then constructed. The increased molecular size of the constructs resulted in improved stability and productivity in E. coli. The oligomeric proteins formed inclusion bodies which were dissolved with dithiothreitol, and the purified apo-metallothioneins were reconstituted with Cd or Zn ions in a reducing condition. The oligomeric hMT proteins incubated with Cd ions showed a typical Cd-thiolate absorbance peak at 245–255 nm. The dimeric and tetrameric hMT proteins exhibited both Cd and Zn binding activities that were respectively two and four times higher than those of the hMT-II monomer protein. These novel oligomeric hMTs may be useful in bioremediation for heavy metals.
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Similar content being viewed by others Explore related subjectsDiscover the latest articles and news from researchers in related subjects, suggested using machine learning. Author information Authors and AffiliationsDepartment of Biotechnology, Graduate School of Engineering, Osaka University, Yamada-oka, Suita, Osaka 565-0871, Japan e-mail: murooka@bio.eng.osaka-u.ac.jp Tel.: +81-6-6879-7416 Fax: +81-6-6879-7418, , , , , , JP
S.-H. Hong, M. Gohya, H. Ono, H. Murakami, M. Yamashita & Y. Murooka
Department of Biological Science and Technology, Tokai University, Nishino, Numazu, Shizuoka 410-32, Japan, , , , , , JP
N. Hirayama
Received: 18 October 1999 / Received last revision: 21 January 2000 / Accepted: 13 February 2000
About this article Cite this articleHong, SH., Gohya, M., Ono, H. et al. Molecular design of novel metal-binding oligomeric human metallothioneins. Appl Microbiol Biotechnol 54, 84–89 (2000). https://doi.org/10.1007/s002530000360
Issue Date: July 2000
DOI: https://doi.org/10.1007/s002530000360
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