Bioaccumulation of heavy metals with protein fusions of metallothionein to bacterial OMPs.
In view of potential biotechnological applications, eukaryotic metallothioneins (MTs) have been expressed in Escherichia coli as fusions to membrane or membrane-associated proteins such as LamB, the peptidoglycan-associated lipoprotein protein (PAL) or a hybrid Lpp/OmpA carrier sequence. The use of different anchors enables the MT moiety to be targeted into various cell compartments thus bringing the metal-binding ability of the resulting hybrids to specific sites of the cell structure. To this end, both full-size and partial sequences of the human or mouse MTs have been genetically fused to: i) the permissive site 153 of the LamB sequence, which loops out the MT to the external medium; ii) the N-terminus of a PAL variant devoid of its N-terminal cystein, which targets expression of the fusion into the periplasm; and iii) the C-terminus of Lpp-OmpA, for anchoring the MT to the outer membrane protein as an N-terminal fusion. Each type of fusion presented a distinct behavior in terms of expression, stability and ability to endow E. coli cells an enhanced accumulation of Cd2+, in good correlation with the number of metal-binding centers contributed by the MT moiety of the fusions. The expression in vivo of metalloproteins bound to bacterial envelope structures opens a way to design biomass with specific metal-binding properties.
Valls
M
M
Departament de Genètica, Universitat de Barcelona, Spain.
González-Duarte
R
R
Atrian
S
S
De Lorenzo
V
V
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D060588:LIM Domain Proteins; D008074:Lipoproteins / Q000378:metabolism; D008668:Metallothionein / Q000378:metabolism*; D019216:Metals, Heavy / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D010457:Peptidoglycan / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins
D001425:Bacterial Outer Membrane Proteins; D003598:Cytoskeletal Proteins; D029968:Escherichia coli Proteins; C031163:ExcC protein, E coli; D060588:LIM Domain Proteins; C103594:LPP protein, human; D008074:Lipoproteins; C491227:Lpp protein, mouse; D019216:Metals, Heavy; D010457:Peptidoglycan; D011506:Proteins; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; C069988:PplA protein, Legionella pneumophila; D008668:Metallothionein