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Abstract An agarase-producing strain NI466 was isolated from Nelson’s Island, Alexandria, Egypt. Based on 16S rRNA analysis, the isolate was identified to be Gayadomonas sp. The effect of different culture conditions on the agarase production was evaluated using Response Surface Methodology (RSM). The highest agarase production was obtained after 14.56 h incubation at 26.12{u00B0}C in a medium consisting of 3% NaCl, 0.18% agar and 14.58g/l tryptone at initial pH 6.0. Enzyme assay using p-nitrophenyl-Ü/Ý-D-galactopyranoside revealed substantial hydrolysis activity toward p-nitrophenyl-Ý-D-galactopyranoside indicating the cleavage of Ý-linkage but not Ü-linkage. Agar hydrolysates produced by the action of the partially purified agarase on agar exhibited a promising anti-oxidative activity |