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    1-Deoxy-D-xylulose 5-phosphate Synthase, the Gene Product of Open Reading Frame (ORF) 2816 and ORF 2895 in Rhodobacter Capsulatus

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    Issue Date
    2001-01-01
    Author
    Hahn, Frederick M.
    Eubanks, Lisa M.
    Testa, Charles A.
    Blagg, Brian S. J.
    Baker, Jonathan A.
    Poulter, C. Dale
    Publisher
    American Society for Microbiology
    Type
    Article
    Article Version
    Scholarly/refereed, publisher version
    Metadata
    Show full item record
    Abstract
    In eubacteria, green algae, and plant chloroplasts, isopentenyl diphosphate, a key intermediate in the biosynthesis of isoprenoids, is synthesized by the methylerythritol phosphate pathway. The five carbons of the basic isoprenoid unit are assembled by joining pyruvate and d-glyceraldehyde 3-phosphate. The reaction is catalyzed by the thiamine diphosphate-dependent enzyme 1-deoxy-d-xylulose 5-phosphate synthase. InRhodobacter capsulatus, two open reading frames (ORFs) carry the genes that encode 1-deoxy-d-xylulose 5-phosphate synthase. ORF 2816 is located in the photosynthesis-related gene cluster, along with most of the genes required for synthesis of the photosynthetic machinery of the bacterium, whereas ORF 2895 is located elsewhere in the genome. The proteins encoded by ORF 2816 and ORF 2895, 1-deoxy-d-xylulose 5-phosphate synthase A and B, containing a His6 tag, were synthesized in Escherichia coli and purified to greater than 95% homogeneity in two steps. 1-Deoxy-d-xylulose 5-phosphate synthase A appears to be a homodimer with 68 kDa subunits. A new assay was developed, and the following steady-state kinetic constants were determined for 1-deoxy-d-xylulose 5-phosphate synthase A and B: Km pyruvate = 0.61 and 3.0 mM, Km d-glyceraldehyde 3-phosphate = 150 and 120 μM, andV max = 1.9 and 1.4 μmol/min/mg in 200 mM sodium citrate (pH 7.4). The ORF encoding 1-deoxy-d-xylulose 5-phosphate synthase B complemented the disrupted essential dxs gene in E. coli strain FH11.
    Description
    This is the publisher's version, also available electronically from "http://jb.asm.org".
    URI
    http://hdl.handle.net/1808/17644
    DOI
    https://doi.org/10.1128/JB.183.1.1-11.2001
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    • Pharmacy Scholarly Works [280]
    Citation
    Hahn, F., Eubanks, L., Testa, C., Blagg, B., Baker, J., & Poulter, C. (2001). 1-Deoxy-D-Xylulose 5-Phosphate Synthase, the Gene Product of Open Reading Frame (ORF) 2816 and ORF 2895 in Rhodobacter capsulatus. Journal of Bacteriology, 183(1), 1-11. http://www.dx.doi.org/10.1128/JB.183.1.1-11.2001

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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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