Functional annotation and structural characterization of a novel lactonase hydrolyzing d-xylono-1,4-lactone-5-phosphate and l -arabino-1,4-lactone-5- phosphate

Magdalena Korczynska, Dao Feng Xiang, Zhening Zhang, Chengfu Xu, Tamari Narindoshvili, Siddhesh S. Kamat, Howard J. Williams, Shawn S. Chang, Peter Kolb, Brandan Hillerich, J. Michael Sauder, Stephen K. Burley, Steven C. Almo, Subramanyam Swaminathan, Brian K. Shoichet, Frank M. Raushel

Research output: Contribution to journalArticle

6 Scopus citations


A novel lactonase from Mycoplasma synoviae 53 (MS53-0025) and Mycoplasma agalactiae PG2 (MAG-6390) was characterized by protein structure determination, molecular docking, gene context analysis, and library screening. The crystal structure of MS53-0025 was determined to a resolution of 2.06 Å. This protein adopts a typical amidohydrolase (β/α)8-fold and contains a binuclear zinc center located at the C-terminal end of the β-barrel. A phosphate molecule was bound in the active site and hydrogen bonds to Lys217, Lys244, Tyr245, Arg275, and Tyr278. Both docking and gene context analysis were used to narrow the theoretical substrate profile of the enzyme, thus directing empirical screening to identify that MS53-0025 and MAG-6390 catalyze the hydrolysis of d-xylono-1,4-lactone-5-phosphate (2) with kcat/Km values of 4.7 × 104 and 5.7 × 104 M-1 s-1 and l-arabino-1,4-lactone- 5-phosphate (7) with kcat/Km values of 1.3 × 10 4 and 2.2 × 104 M-1 s-1, respectively. The identification of the substrate profile of these two phospho-furanose lactonases emerged only when all methods were integrated and therefore provides a blueprint for future substrate identification of highly related amidohydrolase superfamily members.

Original languageEnglish (US)
Pages (from-to)4727-4738
Number of pages12
Issue number28
Publication statusPublished - Jul 22 2014


ASJC Scopus subject areas

  • Biochemistry

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