Isolation, Characterization, Kinetics, and Enzymatic and Nonenzymatic Microbicidal Activities of a Novel c-Type Lysozyme from Plasma of Schistocerca gregaria (Orthoptera: Acrididae) - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Access content directly
Journal Articles Journal of Insect Science Year : 2015

Isolation, Characterization, Kinetics, and Enzymatic and Nonenzymatic Microbicidal Activities of a Novel c-Type Lysozyme from Plasma of Schistocerca gregaria (Orthoptera: Acrididae)

Abstract

A protein, designated as Sgl, showing a muramidase lytic activity to the cell wall of the Gram-positive bacterium Micrococcus lysodeikticus was isolated for the first time from plasma of Escherichia coli-immunized fifth instar Schistocerca gregaria. The isolated Sgl was detected as a single protein band, on both native-and SDS-PAGE, has a molecular weight of similar to 15.7 kDa and an isoelectric point (pI) of ca 9.3 and its antiserum has specifically recognized its isolated form. Fifty-nine percentage of Sgl lytic activity was recovered in the isolated fractions and yielded ca 126-fold increase in specific activity than that of the crude. The partial N-terminal amino acid sequence of the Sgl has 55 and 40% maximum identity with Bombyx mori and Gallus gallus c-type lysozymes, respectively. The antibacterial activity against the Gram-positive and the Gram-negative bacteria were comparatively stronger than that of the hen egg white lysozyme (HEWL). The detected Sgl poration to the inner membrane that reach a maximum ability after 3 h was suggested to operate as a nonenzymatic mechanism for Gram-negative bacterial cell lysis, as tested in a permease-deficient E. coli, ML 35 strain. Sgl showed a maximal muramidase activity at pH 6.2, 30-50 degrees C, and 0.05M Ca2+ or Mg2+; and has a K-m of 0.5 mu g/ml and a V-max of 0.518 with M. lysodeikticus as a substrate. The Sgl displayed a chitinase activity against chitin with a K-m of 0.93 mg/ml and a V-max of 1.63.
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Dates and versions

hal-01165742 , version 1 (19-06-2015)
hal-01165742 , version 2 (21-07-2015)

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Mohamed Elmogy, Taha T. M. Bassal, Hesham A. Yousef, Moataza A. Dorrah, Amr A. Mohamed, et al.. Isolation, Characterization, Kinetics, and Enzymatic and Nonenzymatic Microbicidal Activities of a Novel c-Type Lysozyme from Plasma of Schistocerca gregaria (Orthoptera: Acrididae). Journal of Insect Science, 2015, 15 (1), 10 p. ⟨10.1093/jisesa/iev038⟩. ⟨hal-01165742v2⟩
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