Staphylococcal Surface Display of Metal-Binding Polyhistidyl Peptides

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

Applied and Environmental Microbiology ◽

10.1128/aem.65.3.1092-1098.1999 ◽

1999 ◽

Vol 65 (3) ◽

pp. 1092-1098 ◽

Cited By ~ 125

Author(s):

Pavel Kotrba ◽

Lucie Dolečková ◽

Víctor de Lorenzo ◽

Tomas Ruml

Keyword(s):

Metal Binding ◽

Metal Ion ◽

Rational Design ◽

Binding Capacity ◽

Surface Display ◽

Genetically Engineered ◽

E Coli ◽

Metal Binding Peptides ◽

Binding Peptides ◽

Cell Envelopes

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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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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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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