The focus of this study was to look at relationship between polydispersity caused by high pressure homogenization and molecular weight dependent antimicrobial activity of chitosan. It has been shown that chitosan has antimicrobial activity against bacteria, fungi, and viruses. Chitosan is obtained by partial de-N-acetylation of chitin which, consists of a ß 1-4 copolymer of glucosamine and N-acetylglucosamine residues. In this experiment we compared chitosan of sixteen different molecular weights after being processed through a high pressure homogenizer. Processed chitosan (420 kDa average molecular weight, 30% of acetylation) was dissolved in a 1% (v/v) acetic acid in water to a final concentration of 1% (w/v) and apparent viscosity of 183 MPa. The chitosan solution was passed through a high pressure homogenizer with 0-5 passes at pressure levels 0, 100, 200, and 300 MPa. After processing, the chitosan acetate was investigated to determine the effect on polydispersity in terms of molecular weight and the antimicrobial properties of chitosan at different molecular weights. All compounds were tested against Escherichia coli K-12 to determine antimicrobial activity. There is growing interest in the application of chitosan in food industry due to its wide range of desirable properties including being non-toxic and biodegradability. However, as a hydrophobic material, it is very challenging to work with. Though chitosan is a challenge to work my findings indicated a strong antimicrobial relationship with chitosan at 1% concentration with a molecular weight of 200 kDa and lower against E. coli. In conclusion chitosan has viable application with a variety of foods and can be used as a preservative that decrease bacterial activity below detection level’s and helps to prolong shelf-stable products.