Improved purification, crystallization and crystallographic study of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp. S-77

Noor Dina Muhd Noor, Koji Nishikawa, Hirofumi Nishihara, Ki Seok Yoon, Seiji Ogo, Yoshiki Higuchi

    Research output: Contribution to journalArticlepeer-review

    4 Citations (Scopus)

    Abstract

    The purification procedure of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp. S-77 was improved by applying treatment with trypsin before chromatography. Purified protein samples both with and without trypsin treatment were successfully crystallized using the sitting-drop vapour-diffusion method with polyethylene glycol as a precipitant. Both crystals belonged to space group P21, with unit-cell parameters a = 63.90, b = 118.89, c = 96.70Å, β = 100.61° for the protein subjected to trypsin treatment and a = 65.38, b = 121.45, c = 98.63Å, β = 102.29° for the sample not treated with trypsin. The crystal obtained from the trypsin-treated protein diffracted to 1.60Å resolution, which is considerably better than the 2.00Å resolution obtained without trypsin treatment. The [NiFe]-hydrogenase from Citrobacter sp. S-77 retained catalytic activity with some amount of O2, indicating that it has clear O2 tolerance.

    Original languageEnglish
    Pages (from-to)53-58
    Number of pages6
    JournalActa Crystallographica Section:F Structural Biology Communications
    Volume72
    DOIs
    Publication statusPublished - 2016

    All Science Journal Classification (ASJC) codes

    • Biophysics
    • Structural Biology
    • Biochemistry
    • Genetics
    • Condensed Matter Physics

    Fingerprint

    Dive into the research topics of 'Improved purification, crystallization and crystallographic study of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp. S-77'. Together they form a unique fingerprint.

    Cite this