Mucor rouxii was selected due to the high biomass production and significant quantities of chitosan in its cell walls. M. rouxii cultured in Potato dextrose Broth for 96 hrs. at 30 C◦ in a shaking incubator at 150 rpm and 5.5 pH, then the fungal mycelial were dried, grounded and weighted. Mycelial dry weight in total was 68.8g with a yield of 1.72 ± 0.25 g/500ml, chitosan was extracted using the classic chemical method followed by precipitation of chitosan by using sodium hydroxide. chitosan yield was 2.13%, The degree of deacetylation of chitosan extracted from M. rouxii was 82.22% with low Molecular weight 63.67 kDa. The Antimicrobial properties of extracted chitosan was studded on four pathogenic bacteria by MIC method the most resistant strains which were S. aureus, whereas the most vulnerable strains were A. baumannii and E. coli. to produce natural chitosan and replace old sources (crustaceans). The observed antimicrobial properties also indicate an acceptable effect of chitosan on some strains that needs further study.