Enhanced Bioaccumulation of Heavy Metal Ions by Bacterial Cells Due to Surface Display of Short Metal Binding Peptides

ABSTRACT Metal binding peptides of sequences Gly-His-His-Pro-His-Gly (named HP) and Gly-Cys-Gly-Cys-Pro-Cys-Gly-Cys-Gly (named CP) were genetically engineered into LamB protein and expressed in Escherichia coli. The Cd2+-to-HP and Cd2+-to-CP stoichiometries of peptides were 1:1 and 3:1, respectively. Hybrid LamB proteins were found to be properly folded in the outer membrane ofE. coli. Isolated cell envelopes of E. colibearing newly added metal binding peptides showed an up to 1.8-fold increase in Cd2+ binding capacity. The bioaccumulation of Cd2+, Cu2+, and Zn2+ by E. coli was evaluated. Surface display of CP multiplied the ability of E. coli to bind Cd2+ from growth medium fourfold. Display of HP peptide did not contribute to an increase in the accumulation of Cu2+ and Zn2+. However, Cu2+ ceased contribution of HP for Cd2+accumulation, probably due to the strong binding of Cu2+ to HP. Thus, considering the cooperation of cell structures with inserted peptides, the relative affinities of metal binding peptide and, for example, the cell wall to metal ion should be taken into account in the rational design of peptide sequences possessing specificity for a particular metal.

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Staphylococcal Surface Display of Metal-Binding Polyhistidyl Peptides

Applied and Environmental Microbiology ◽

10.1128/aem.66.3.1243-1248.2000 ◽

2000 ◽

Vol 66 (3) ◽

pp. 1243-1248 ◽

Cited By ~ 78

Author(s):

Patrik Samuelson ◽

Henrik Wernérus ◽

Malin Svedberg ◽

Stefan Ståhl

Keyword(s):

Metal Binding ◽

Binding Capacity ◽

Surface Display ◽

Surface Proteins ◽

Chimeric Proteins ◽

Divalent Metal Ions ◽

Gram Positive Bacteria ◽

Metal Binding Peptides ◽

Binding Peptides ◽

First Time

ABSTRACT Recombinant Staphylococcus xylosus andStaphylococcus carnosus strains were generated with surface-exposed chimeric proteins containing polyhistidyl peptides designed for binding to divalent metal ions. Surface accessibility of the chimeric surface proteins was demonstrated and the chimeric surface proteins were found to be functional in terms of metal binding, since the recombinant staphylococcal cells were shown to have gained Ni2+- and Cd2+-binding capacity, suggesting that such bacteria could find use in bioremediation of heavy metals. This is, to our knowledge, the first time that recombinant, surface-exposed metal-binding peptides have been expressed on gram-positive bacteria. Potential environmental or biosensor applications for such recombinant staphylococci as biosorbents are discussed.

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Bacterial surface display of metal binding peptides as whole-cell biocatalysts for 4-nitroaniline reduction

RSC Advances ◽

10.1039/c5ra18561k ◽

2015 ◽

Vol 5 (107) ◽

pp. 87998-88001 ◽

Cited By ~ 3

Author(s):

Dong-Yu Tsai ◽

Yi-Jung Tsai ◽

Chia-Ho Yen ◽

Chun-Yu Ouyang ◽

Yi-Chun Yeh

Keyword(s):

Metal Binding ◽

Surface Display ◽

Whole Cell ◽

Bacterial Surface ◽

Bacterial Surface Display ◽

Metal Binding Peptides ◽

Recombinant Microorganisms ◽

Binding Peptides

Using recombinant microorganisms expressing metal binding peptides as whole-cell biocatalysts for 4-nitroaniline reduction.

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Genetic Engineering of Escherichia coli for Enhanced Uptake and Bioaccumulation of Mercury

Applied and Environmental Microbiology ◽

10.1128/aem.67.11.5335-5338.2001 ◽

2001 ◽

Vol 67 (11) ◽

pp. 5335-5338 ◽

Cited By ~ 87

Author(s):

Weon Bae ◽

Rajesh K. Mehra ◽

Ashok Mulchandani ◽

Wilfred Chen

Keyword(s):

Heavy Metals ◽

Metal Binding ◽

Heavy Metal Contamination ◽

Genetically Engineered ◽

Transport Systems ◽

Limiting Factor ◽

Binding Motif ◽

Metal Binding Peptides ◽

Binding Peptides ◽

Genetically Engineered Organisms

ABSTRACT Synthetic phytochelatins (ECs) are a new class of metal-binding peptides with a repetitive metal-binding motif, (Glu-Cys) n Gly, which were shown to bind heavy metals more effectively than metallothioneins. However, the limited uptake across the cell membrane is often the rate-limiting factor for the intracellular bioaccumulation of heavy metals by genetically engineered organisms expressing these metal-binding peptides. In this paper, two potential solutions were investigated to overcome this uptake limitation either by coexpressing an Hg2+ transport system with (Glu-Cys)20Gly (EC20) or by directly expressing EC20 on the cell surface. Both approaches were equally effective in increasing the bioaccumulation of Hg2+. Since the available transport systems are presently limited to only a few heavy metals, our results suggest that bioaccumulation by bacterial sorbents with surface-expressed metal-binding peptides may be useful as a universal strategy for the cleanup of heavy metal contamination.

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Heavy Metal-Binding Peptides and Proteins in Plants. A Review

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19991057 ◽

1999 ◽

Vol 64 (7) ◽

pp. 1057-1086 ◽

Cited By ~ 44

Author(s):

Pavel Kotrba ◽

Tomáš Macek ◽

Tomáš Ruml

Keyword(s):

Heavy Metal ◽

Metal Binding ◽

Metal Ion ◽

Plant Pathogens ◽

General Structure ◽

Transition Metal Ions ◽

Stress Factors ◽

Sucrose Starvation ◽

Metal Binding Peptides ◽

Binding Peptides

In plants, two kinds of specific metal-binding peptides or proteins are synthesized. Plant metallothioneins (MTs) and MT-like proteins are cysteine-rich translation products of genes inducible in tissue-specific manner during embryogenesis and plant development. In addition, differential expression of MT-like protein genes could be due to variation of external heavy metal concentrations (especially of Cu2+ and Fe2+), influence of various stress factors (heat shock, sucrose starvation, oxidative stress, wounding, plant pathogens). The principal role of plant MTs and MT-like proteins seems to be in homeostasis of essential transition metals rather than in metal detoxification. Phytochelatins (PCs) have general structure (γ-Glu-Cys)n-Xaa, where n = 2-11 and Xaa amino acids Gly, β-Ala, Ser, and Glu which depend on the species; the des-Xaa forms of PC also exist. PCs are synthesized in plants and some yeasts by a constitutive enzyme phytochelatin synthase (active only in the presence of free heavy metal ion) from glutathione or its anologue. Despite the PC capability of forming complexes with transition metal ions (their role in metal homeostasis could not be excluded) and virtually prominent role in Cd2+ detoxification within plant cell, there is no evidence that elevated production of PCs may contribute to differential tolerance and/or could be responsible for the resistance to toxic metals. A review with 172 references.

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Thermochemical and conformational studies of Ni(II) and Zn(II) ternary complexes of alternative metal binding peptides with nitrilotriacetic acid

International Journal of Mass Spectrometry ◽

10.1016/j.ijms.2021.116792 ◽

2022 ◽

pp. 116792

Author(s):

Amber A. Flores ◽

Anna V. Arredondo ◽

Anna J. Corrales ◽

Chloe L. Duvak ◽

Charles L. Mitchell ◽

...

Keyword(s):

Metal Binding ◽

Nitrilotriacetic Acid ◽

Ternary Complexes ◽

Conformational Studies ◽

Metal Binding Peptides ◽

Binding Peptides

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A new approach to biomining: Bioengineering surfaces for metal recovery from aqueous solutions

Scientific Reports ◽

10.1038/s41598-019-52778-2 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 3

Author(s):

Jesica Urbina ◽

Advait Patil ◽

Kosuke Fujishima ◽

Ivan G. Paulino-Lima ◽

Chad Saltikov ◽

...

Keyword(s):

Aqueous Solutions ◽

Metal Binding ◽

Raw Materials ◽

Extraction Methods ◽

Metal Recovery ◽

Consumer Electronics ◽

Metal Extraction ◽

Copper Binding ◽

Metal Binding Peptides ◽

Binding Peptides

Abstract Electronics waste production has been fueled by economic growth and the demand for faster, more efficient consumer electronics. The glass and metals in end-of-life electronics components can be reused or recycled; however, conventional extraction methods rely on energy-intensive processes that are inefficient when applied to recycling e-waste that contains mixed materials and small amounts of metals. To make e-waste recycling economically viable and competitive with obtaining raw materials, recovery methods that lower the cost of metal reclamation and minimize environmental impact need to be developed. Microbial surface adsorption can aid in metal recovery with lower costs and energy requirements than traditional metal-extraction approaches. We introduce a novel method for metal recovery by utilizing metal-binding peptides to functionalize fungal mycelia and enhance metal recovery from aqueous solutions such as those found in bioremediation or biomining processes. Using copper-binding as a proof-of-concept, we compared binding parameters between natural motifs and those derived in silico, and found comparable binding affinity and specificity for Cu. We then combined metal-binding peptides with chitin-binding domains to functionalize a mycelium-based filter to enhance metal recovery from a Cu-rich solution. This finding suggests that engineered peptides could be used to functionalize biological surfaces to recover metals of economic interest and allow for metal recovery from metal-rich effluent with a low environmental footprint, at ambient temperatures, and under circumneutral pH.

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Generation of Metal-Binding Staphylococci through Surface Display of Combinatorially Engineered Cellulose-Binding Domains

Applied and Environmental Microbiology ◽

10.1128/aem.67.10.4678-4684.2001 ◽

2001 ◽

Vol 67 (10) ◽

pp. 4678-4684 ◽

Cited By ~ 32

Author(s):

Henrik Wernérus ◽

Janne Lehtiö ◽

Tuula Teeri ◽

Per-Åke Nygren ◽

Stefan Ståhl

Keyword(s):

Metal Binding ◽

Binding Capacity ◽

Surface Display ◽

Surface Proteins ◽

Chimeric Proteins ◽

Phage Display Technology ◽

Binding Domains ◽

Combinatorial Protein Engineering ◽

Display Technology ◽

Cellulose Binding

ABSTRACT Ni2+-binding staphylococci were generated through surface display of combinatorially engineered variants of a fungal cellulose-binding domain (CBD) from Trichoderma reeseicellulase Cel7A. Novel CBD variants were generated by combinatorial protein engineering through the randomization of 11 amino acid positions, and eight potentially Ni2+-binding CBDs were selected by phage display technology. These new variants were subsequently genetically introduced into chimeric surface proteins for surface display on Staphylococcus carnosus cells. The expressed chimeric proteins were shown to be properly targeted to the cell wall of S. carnosus cells, since full-length proteins could be extracted and affinity purified. Surface accessibility for the chimeric proteins was demonstrated, and furthermore, the engineered CBDs, now devoid of cellulose-binding capacity, were shown to be functional with regard to metal binding, since the recombinant staphylococci had gained Ni2+-binding capacity. Potential environmental applications for such tailor-made metal-binding bacteria as bioadsorbents in biofilters or biosensors are discussed.

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Identification of Metal-Binding Peptides and Their Conjugation onto Nanoparticles of Superparamagnetic Iron Oxides and Liposomes

ACS Applied Materials & Interfaces ◽

10.1021/acsami.0c06138 ◽

2020 ◽

Vol 12 (22) ◽

pp. 24623-24634

Author(s):

Tomonobu Kodama ◽

Akifumi Yoshihara ◽

Isha Goel ◽

Masaki Sekino ◽

Akihiro Kuwahata ◽

...

Keyword(s):

Iron Oxides ◽

Metal Binding ◽

Superparamagnetic Iron Oxides ◽

Metal Binding Peptides ◽

Binding Peptides

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Autodisplay: Functional Display of Active β-Lactamase on the Surface of Escherichia coli by the AIDA-I Autotransporter

Journal of Bacteriology ◽

10.1128/jb.182.13.3726-3733.2000 ◽

2000 ◽

Vol 182 (13) ◽

pp. 3726-3733 ◽

Cited By ~ 79

Author(s):

Claus T. Lattemann ◽

Jochen Maurer ◽

Elke Gerland ◽

Thomas F. Meyer

Keyword(s):

Escherichia Coli ◽

Fusion Protein ◽

Surface Display ◽

Heterologous Proteins ◽

Bacterial Surface ◽

E Coli ◽

C Terminus ◽

Protease Treatment ◽

Efficient Expression ◽

Cell Envelopes

ABSTRACT Members of the protein family of immunoglobulin A1 protease-like autotransporters comprise multidomain precursors consisting of a C-terminal autotransporter domain that promotes the translocation of N-terminally attached passenger domains across the cell envelopes of gram-negative bacteria. Several autotransporter domains have recently been shown to efficiently promote the export of heterologous passenger domains, opening up an effective tool for surface display of heterologous proteins. Here we report on the autotransporter domain of the Escherichia coli adhesin involved in diffuse adherence (AIDA-I), which was genetically fused to the C terminus of the periplasmic enzyme β-lactamase, leading to efficient expression of the fusion protein in E. coli. The β-lactamase moiety of the fusion protein was presented on the bacterial surface in a stable manner, and the surface-located β-lactamase was shown to be enzymatically active. Enzymatic activity was completely removed by protease treatment, indicating that surface display of β-lactamase was almost quantitative. The periplasmic domain of the outer membrane protein OmpA was not affected by externally added proteases, demonstrating that the outer membranes of E. coli cells expressing the β-lactamase AIDA-I fusion protein remained physiologically intact.

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