Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies

doi:10.3389/fmicb.2015.00752

Title: Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies

Article Details
Other Related Research

Multiheme cytochromes have been implicated in Geobacter sulfurreducens extracellular electron transfer (EET). These proteins are potential targets to improve EET and enhance bioremediation and electrical current production by G. sulfurreducens. However, the functional characterization of multiheme cytochromes is particularly complex due to the co-existence of several microstates in solution, connecting the fully reduced and fully oxidized states. Throughout the last decade, new strategies have been developed to characterize multiheme redox proteins functionally and structurally. These strategies were used to reveal the functional mechanism of G. sulfurreducens multiheme cytochromes and also to identify key residues in these proteins for EET. In previous studies, we set the foundations for enhancement of the EET abilities of G. sulfurreducens by characterizing a family of five triheme cytochromes (PpcA-E). These periplasmic cytochromes are implicated in electron transfer between the oxidative reactions of metabolism in the cytoplasm and the reduction of extracellular terminal electron acceptors at the cell's outer surface. The results obtained suggested that PpcA can couple e ^-/H ⁺ transfer, a property that might contribute to the proton electrochemical gradient across the cytoplasmic membrane for metabolic energy production. The structural and functional properties of PpcA were characterized in detail and used for rational designmore »

Authors:

Dantas, Joana M. ^[1] ; Morgado, Leonor ^[1] ; Aklujkar, Muktak ^[2] ; Bruix, Marta ^[3] ; Londer, Yuri Y. ^[4] ; Schiffer, Marianne ^[4] ; Pokkuluri, P. Raj ^[4] ; Salgueiro, Carlos A. ^[1]

Univ. Nova de Lisboa, Caparica (Portugal)
Towson Univ., Towson, MD (United States)
Instituto de Quimica-Fisica "Rocasolano", Madrid (Spain)
Argonne National Lab. (ANL), Lemont, IL (United States)

Publication Date:: 2015-07-30

Grant/Contract Number:: AC02-06CH11357

Type:: Accepted Manuscript

Journal Name:: Frontiers in Microbiology

Additional Journal Information:: Journal Volume: 6; Journal ID: ISSN 1664-302X

Publisher:: Frontiers Research Foundation

Research Org:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Org:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Country of Publication:: United States

Language:: English

Subject:: Geobacter; cytochrome c; multiheme; extracellular electron transfer; Geobacter mutant strains

OSTI Identifier:: 1224905


                    Dantas, Joana M., Morgado, Leonor, Aklujkar, Muktak, Bruix, Marta, Londer, Yuri Y., Schiffer, Marianne, Pokkuluri, P. Raj, and Salgueiro, Carlos A.. Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies.  United States: N. p.,  
        Web.  doi:10.3389/fmicb.2015.00752.

Copy to clipboard


                    Dantas, Joana M., Morgado, Leonor, Aklujkar, Muktak, Bruix, Marta, Londer, Yuri Y., Schiffer, Marianne, Pokkuluri, P. Raj, & Salgueiro, Carlos A.. Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies.  United States.  doi:10.3389/fmicb.2015.00752.

Copy to clipboard


                    Dantas, Joana M., Morgado, Leonor, Aklujkar, Muktak, Bruix, Marta, Londer, Yuri Y., Schiffer, Marianne, Pokkuluri, P. Raj, and Salgueiro, Carlos A.. 2015.  
        "Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies".  United States.  
        doi:10.3389/fmicb.2015.00752.  https://www.osti.gov/servlets/purl/1224905.

Copy to clipboard


                    
@article{osti_1224905,

  title        = {Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies},

  author       = {Dantas, Joana M. and Morgado, Leonor and Aklujkar, Muktak and Bruix, Marta and Londer, Yuri Y. and Schiffer, Marianne and Pokkuluri, P. Raj and Salgueiro, Carlos A.},

  abstractNote = {Multiheme cytochromes have been implicated in Geobacter sulfurreducens extracellular electron transfer (EET). These proteins are potential targets to improve EET and enhance bioremediation and electrical current production by G. sulfurreducens. However, the functional characterization of multiheme cytochromes is particularly complex due to the co-existence of several microstates in solution, connecting the fully reduced and fully oxidized states. Throughout the last decade, new strategies have been developed to characterize multiheme redox proteins functionally and structurally. These strategies were used to reveal the functional mechanism of G. sulfurreducens multiheme cytochromes and also to identify key residues in these proteins for EET. In previous studies, we set the foundations for enhancement of the EET abilities of G. sulfurreducens by characterizing a family of five triheme cytochromes (PpcA-E). These periplasmic cytochromes are implicated in electron transfer between the oxidative reactions of metabolism in the cytoplasm and the reduction of extracellular terminal electron acceptors at the cell's outer surface. The results obtained suggested that PpcA can couple e-/H+ transfer, a property that might contribute to the proton electrochemical gradient across the cytoplasmic membrane for metabolic energy production. The structural and functional properties of PpcA were characterized in detail and used for rational design of a family of 23 single site PpcA mutants. In this review, we summarize the functional characterization of the native and mutant proteins. Mutants that retain the mechanistic features of PpcA and adopt preferential e-/H+ transfer pathways at lower reduction potential values compared to the wild-type protein were selected for in vivo studies as the best candidates to increase the electron transfer rate of G. sulfurreducens. For the first time G. sulfurreducens strains have been manipulated by the introduction of mutant forms of essential proteins with the aim to develop and improve bioelectrochemical technologies.},

  doi          = {10.3389/fmicb.2015.00752},

  journal      = {Frontiers in Microbiology},

  number       = ,

  volume       = 6,

  place        = {United States},

  year         = {2015},

  month        = {7}

}

Copy to clipboard

Free Publicly Accessible Full Text
Accepted Manuscript (DOE)
Publisher's Version of Record
10.3389/fmicb.2015.00752
https://doi.org/10.3389/fmicb.2015.00752

Citation Metrics
Cited by: 5works
Citation information provided by
Web of Science

Save / Share this Record
Export Metadata
- Endnote
- RIS
- Excel
- CSV
- XML
Save to My Library
Send to Email

Send to Email

Email address:

Content:

Similar Records

Similar records in DOE PAGES and OSTI.GOV collections:

Electron Paramagnetic Resonance Characterization of the Triheme Cytochrome from Geobacter sulfurreducens

Journal Article Ponomarenko, Nina ; Niklas, Jens ; Pokkuluri, P. Raj ; ... January 2018 - Biochemistry

Periplasmic cytochrome A (PpcA) is a representative of a broad class of multiheme cytochromes functioning as protein “nanowires” for storage and extracellular transfer of multiple electrons in the δ-proteobacterium Geobacter sulfurreducens. PpcA contains three bis-His coordinated hemes held in a spatial arrangement that is highly conserved among the multiheme cytochromes c ₃ and c ₇ families, carries low potential hemes, and is notable for having one of the lowest number of amino acids utilized to maintain a characteristic protein fold and site-specific heme function. Low temperature X-band electron paramagnetic resonance (EPR) spectroscopy has been used to characterize the electronic configurationmore »« less
A trans-outer membrane porin-cytochrome protein complex for extracellular electron transfer by Geobacter sulfurreducens PCA

Journal Article Liu, Yimo ; Wang, Zheming ; Liu, Juan ; ... September 2014 - Environmental Microbiology Reports

The multiheme, outer membrane c-type cytochrome (c-Cyt) OmcB of Geobacter sulfurreducens was previously proposed to mediate electron transfer across the outer membrane. However, the underlying mechanism has remained uncharacterized. In G. sulfurreducens, the omcB gene is part of two tandem four-gene clusters, each is predicted to encode a transcriptional factor (OrfR/OrfS), a porin-like outer membrane protein (OmbB/OmbC), a periplasmic c-type cytochrome (OmaB/OmaC), and an outer membrane c-Cyt (OmcB/OmcC), respectively. Here we showed that OmbB/OmbC, OmaB/OmaC and OmcB/OmcC of G. sulfurreducens PCA formed the porin-cytochrome (Pcc) protein complexes, which were involved in transferring electrons across the outer membrane. The isolated Pccmore »« less
Cited by 39Full Text Available
Bidirectional Photoinduced Electron Transfer in Ruthenium(II)-Tris-bipyridyl-Modified PpcA, a Multi-heme c -Type Cytochrome from Geobacter sulfurreducens

Journal Article - in OSTI.gov collection Kokhan, Oleksandr ; Ponomarenko, Nina S. ; Pokkuluri, P. Raj ; ... June 2015 - Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry

PpcA, a tri-heme cytochrome c7 from Geobacter sulfurreducens was investigated as a model for photosensitizer-initiated electron transfer within a multi-heme "molecular wire" protein architecture. E. coli expression of PpcA was found to be tolerant of cysteine site-directed mutagenesis, demonstrated by the successful expression of natively folded proteins bearing cysteine mutations at a series of sites selected to vary characteristically with respect to the three -CXXCH- heme binding domains. A preliminary survey of 5 selected mutants found that the introduced cysteines can be readily covalently linked to a Ru(II)-(2,2'-bpy)2(4-bromomethyl-4’-methyl-2,2'-bpy) photosensitizer (where bpy = bipyridine), and that the linked constructs support bothmore »« less
Full Text Available
Global Proteome Response to Deletion of Genes Related to Mercury Methylation and Dissimilatory Metal Reduction Reveals Changes in Respiratory Metabolism in Geobacter sulfurreducens PCA

Journal Article Qian, Chen ; Johs, Alexander ; Chen, Hongmei ; ... July 2016 - Journal of Proteome Research

Geobacter sulfurreducens PCA can reduce, sorb, and methylate mercury (Hg); however, the underlying biochemical mechanisms of these processes and interdependent metabolic pathways remain unknown. In this study, shotgun proteomics was used to compare global proteome profiles between wild-type G. sulfurreducens PCA and two mutant strains: a ΔhgcAB mutant, which is deficient in two genes known to be essential for Hg methylation and a ΔomcBESTZ mutant, which is deficient in five outer membrane c-type cytochromes and thus impaired in its ability for dissimilatory metal ion reduction. We were able to delineate the global response of G. sulfurreducens PCA in both mutantsmore »« less
Cited by 3Full Text Available
Scarless genome editing and stable inducible expression vectors for Geobacter sulfurreducens

Journal Article Chan, Chi Ho ; Levar, Caleb E. ; Zacharoff, Lori ; ... August 2015 - Applied and Environmental Microbiology

Metal reduction by members of the Geobacteraceae is encoded by multiple gene clusters, and the study of extracellular electron transfer often requires biofilm development on surfaces. Genetic tools that utilize polar antibiotic cassette insertions limit mutant construction and complementation. In addition, unstable plasmids create metabolic burdens that slow growth, and the presence of antibiotics such as kanamycin can interfere with the rate and extent of Geobacter biofilm growth. We report here genetic system improvements for the model anaerobic metal-reducing bacterium Geobacter sulfurreducens. A motile strain of G. sulfurreducens was constructed by precise removal of a transposon interrupting the fgrM flagellarmore »« less
Cited by 5Full Text Available

Access journal articles and accepted manuscripts resulting from DOE research funding

Title: Rational engineering of Geobacter sulfurreducens electron transfer components: A foundation for building improved Geobacter-based bioelectrochemical technologies

Access journal articles and accepted manuscripts
resulting from DOE research funding