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  1. Article ; Online: Genomic characterization of rare earth binding by Shewanella oneidensis.

    Medin, Sean / Schmitz, Alexa M / Pian, Brooke / Mini, Kuunemuebari / Reid, Matthew C / Holycross, Megan / Gazel, Esteban / Wu, Mingming / Barstow, Buz

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 15975

    Abstract: Rare earth elements (REE) are essential ingredients of sustainable energy technologies, but separation of individual REE is one of the hardest problems in chemistry today. Biosorption, where molecules adsorb to the surface of biological materials, offers ...

    Abstract Rare earth elements (REE) are essential ingredients of sustainable energy technologies, but separation of individual REE is one of the hardest problems in chemistry today. Biosorption, where molecules adsorb to the surface of biological materials, offers a sustainable alternative to environmentally harmful solvent extractions currently used for separation of rare earth elements (REE). The REE-biosorption capability of some microorganisms allows for REE separations that, under specialized conditions, are already competitive with solvent extractions, suggesting that genetic engineering could allow it to leapfrog existing technologies. To identify targets for genomic improvement we screened 3,373 mutants from the whole genome knockout collection of the known REE-biosorbing microorganism Shewanella oneidensis MR-1. We found 130 genes that increased biosorption of the middle REE europium, and 112 that reduced it. We verified biosorption changes from the screen for a mixed solution of three REE (La, Eu, Yb) using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in solution conditions with a range of ionic strengths and REE concentrations. We identified 18 gene ontologies and 13 gene operons that make up key systems that affect biosorption. We found, among other things, that disruptions of a key regulatory component of the arc system (hptA), which regulates cellular response to anoxic environments and polysaccharide biosynthesis related genes (wbpQ, wbnJ, SO_3183) consistently increase biosorption across all our solution conditions. Our largest total biosorption change comes from our SO_4685, a capsular polysaccharide (CPS) synthesis gene, disruption of which results in an up to 79% increase in biosorption; and nusA, a transcriptional termination/anti-termination protein, disruption of which results in an up to 35% decrease in biosorption. Knockouts of glnA, pyrD, and SO_3183 produce small but significant increases (≈ 1%) in relative biosorption affinity for ytterbium over lanthanum in multiple solution conditions tested, while many other genes we explored have more complex binding affinity changes. Modeling suggests that while these changes to lanthanide biosorption selectivity are small, they could already reduce the length of repeated enrichment process by up to 27%. This broad exploratory study begins to elucidate how genetics affect REE-biosorption by S. oneidensis, suggests new areas of investigation for better mechanistic understanding of the membrane chemistry involved in REE binding, and offer potential targets for improving biosorption and separation of REE by genetic engineering.
    MeSH term(s) Genomics ; Shewanella/genetics ; Europium ; Solvents
    Chemical Substances Europium (444W947O8O) ; Solvents
    Language English
    Publishing date 2023-09-25
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-42742-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Single-cell multimodal imaging uncovers energy conversion pathways in biohybrids.

    Fu, Bing / Mao, Xianwen / Park, Youngchan / Zhao, Zhiheng / Yan, Tianlei / Jung, Won / Francis, Danielle H / Li, Wenjie / Pian, Brooke / Salimijazi, Farshid / Suri, Mokshin / Hanrath, Tobias / Barstow, Buz / Chen, Peng

    Nature chemistry

    2023  Volume 15, Issue 10, Page(s) 1400–1407

    Abstract: Microbe-semiconductor biohybrids, which integrate microbial enzymatic synthesis with the light-harvesting capabilities of inorganic semiconductors, have emerged as promising solar-to-chemical conversion systems. Improving the electron transport at the ... ...

    Abstract Microbe-semiconductor biohybrids, which integrate microbial enzymatic synthesis with the light-harvesting capabilities of inorganic semiconductors, have emerged as promising solar-to-chemical conversion systems. Improving the electron transport at the nano-bio interface and inside cells is important for boosting conversion efficiencies, yet the underlying mechanism is challenging to study by bulk measurements owing to the heterogeneities of both constituents. Here we develop a generalizable, quantitative multimodal microscopy platform that combines multi-channel optical imaging and photocurrent mapping to probe such biohybrids down to single- to sub-cell/particle levels. We uncover and differentiate the critical roles of different hydrogenases in the lithoautotrophic bacterium Ralstonia eutropha for bioplastic formation, discover this bacterium's surprisingly large nanoampere-level electron-uptake capability, and dissect the cross-membrane electron-transport pathways. This imaging platform, and the associated analytical framework, can uncover electron-transport mechanisms in various types of biohybrid, and potentially offers a means to use and engineer R. eutropha for efficient chemical production coupled with photocatalytic materials.
    MeSH term(s) Electron Transport ; Multimodal Imaging
    Language English
    Publishing date 2023-07-27
    Publishing country England
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2464596-5
    ISSN 1755-4349 ; 1755-4330
    ISSN (online) 1755-4349
    ISSN 1755-4330
    DOI 10.1038/s41557-023-01285-z
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Unusual 1-3 peptidoglycan cross-links in Acetobacteraceae are made by L,D-transpeptidases with a catalytic domain distantly related to YkuD domains.

    Alamán-Zárate, Marcel G / Rady, Brooks J / Evans, Caroline A / Pian, Brooke / Greetham, Darren / Marecos-Ortiz, Sabrina / Dickman, Mark J / Lidbury, Ian D E A / Lovering, Andrew L / Barstow, Buz M / Mesnage, Stéphane

    The Journal of biological chemistry

    2023  Volume 300, Issue 1, Page(s) 105494

    Abstract: Peptidoglycan is an essential component of the bacterial cell envelope that contains glycan chains substituted by short peptide stems. Peptide stems are polymerized by D,D-transpeptidases, which make bonds between the amino acid in position four of a ... ...

    Abstract Peptidoglycan is an essential component of the bacterial cell envelope that contains glycan chains substituted by short peptide stems. Peptide stems are polymerized by D,D-transpeptidases, which make bonds between the amino acid in position four of a donor stem and the third residue of an acceptor stem (4-3 cross-links). Some bacterial peptidoglycans also contain 3-3 cross-links that are formed by another class of enzymes called L,D-transpeptidases which contain a YkuD catalytic domain. In this work, we investigate the formation of unusual bacterial 1-3 peptidoglycan cross-links. We describe a version of the PGFinder software that can identify 1-3 cross-links and report the high-resolution peptidoglycan structure of Gluconobacter oxydans (a model organism within the Acetobacteraceae family). We reveal that G. oxydans peptidoglycan contains peptide stems made of a single alanine as well as several dipeptide stems with unusual amino acids at their C-terminus. Using a bioinformatics approach, we identified a G. oxydans mutant from a transposon library with a drastic reduction in 1-3 cross-links. Through complementation experiments in G. oxydans and recombinant protein production in a heterologous host, we identify an L,D-transpeptidase enzyme with a domain distantly related to the YkuD domain responsible for these non-canonical reactions. This work revisits the enzymatic capabilities of L,D-transpeptidases, a versatile family of enzymes that play a key role in bacterial peptidoglycan remodelling.
    MeSH term(s) Amino Acids/genetics ; Bacterial Proteins/chemistry ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Catalytic Domain/genetics ; Peptidoglycan/chemistry ; Peptidoglycan/genetics ; Peptidoglycan/metabolism ; Peptidyl Transferases/chemistry ; Peptidyl Transferases/genetics ; Peptidyl Transferases/metabolism ; Software ; Gluconobacter oxydans/enzymology ; Gluconobacter oxydans/genetics ; Computational Biology ; Genetic Complementation Test ; Models, Molecular ; Protein Structure, Tertiary
    Chemical Substances Amino Acids ; Bacterial Proteins ; Peptidoglycan ; Peptidyl Transferases (EC 2.3.2.12)
    Language English
    Publishing date 2023-11-23
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1016/j.jbc.2023.105494
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Generation of a Gluconobacter oxydans knockout collection for improved extraction of rare earth elements.

    Schmitz, Alexa M / Pian, Brooke / Medin, Sean / Reid, Matthew C / Wu, Mingming / Gazel, Esteban / Barstow, Buz

    Nature communications

    2021  Volume 12, Issue 1, Page(s) 6693

    Abstract: Bioleaching of rare earth elements (REEs), using microorganisms such as Gluconobacter oxydans, offers a sustainable alternative to environmentally harmful thermochemical extraction, but is currently not very efficient. Here, we generate a whole-genome ... ...

    Abstract Bioleaching of rare earth elements (REEs), using microorganisms such as Gluconobacter oxydans, offers a sustainable alternative to environmentally harmful thermochemical extraction, but is currently not very efficient. Here, we generate a whole-genome knockout collection of single-gene transposon disruption mutants for G. oxydans B58, to identify genes affecting the efficacy of REE bioleaching. We find 304 genes whose disruption alters the production of acidic biolixiviant. Disruption of genes underlying synthesis of the cofactor pyrroloquinoline quinone (PQQ) and the PQQ-dependent membrane-bound glucose dehydrogenase nearly eliminates bioleaching. Disruption of phosphate-specific transport system genes enhances bioleaching by up to 18%. Our results provide a comprehensive roadmap for engineering the genome of G. oxydans to further increase its bioleaching efficiency.
    MeSH term(s) Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Gene Knockout Techniques/methods ; Genetic Engineering/methods ; Genome, Bacterial/genetics ; Gluconobacter oxydans/genetics ; Gluconobacter oxydans/metabolism ; Glucose Dehydrogenases/genetics ; Glucose Dehydrogenases/metabolism ; Industrial Microbiology/methods ; Metals, Rare Earth/metabolism ; PQQ Cofactor/genetics ; PQQ Cofactor/metabolism ; Reproducibility of Results
    Chemical Substances Bacterial Proteins ; Metals, Rare Earth ; PQQ Cofactor (72909-34-3) ; Glucose Dehydrogenases (EC 1.1.1.-) ; glucose dehydrogenase (pyrroloquinoline-quinone) (EC 1.1.5.2)
    Language English
    Publishing date 2021-11-18
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-021-27047-4
    Database MEDical Literature Analysis and Retrieval System OnLINE

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