Surfactants are surface-active materials with numerous applications, including: detergents and soaps, cosmetics, fabric softeners, pharmaceutical creams and ointments, and plasticizers. Currently, amino acid-based surfactants have generated interest as antibacterial agents, chiral resolution, chelating agents, and as switchable surfactants. Furthermore, these surfactants are of particular interest because they are largely biodegradable, due to a relatively labile ester or amide bond. We describe the synthesis of O-acylated amino acid surfactants with hydrophobic ω-alkenyl chains. Amino acids, such as L-Leucine and L-Phenylalanine, were acylated with ω-undecylenyl alcohol using a Fischer esterification reaction. The amino-acid based surfactants were then purified using silica-gel chromatography, as well as an amine-selective resin. Afterwards, these two types of amino acid-based surfactants were evaluated for their antimicrobial activity by initially performing a Kirby-Bauer Assay, using Mueller-Hinton Agar (MHA) plates to observe disc diffusion rates based on the susceptibility of Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive) bacteria. Recently, further antimicrobial testing was conducted using a different methodology known as a Micro Broth Dilution Assay, in order to determine the Minimum Inhibitory Concentrations (MICs) of both Escherichia coli and Staphylococcus aureus bacteria for each of the amino-acid based surfactants that have been synthesized. The results of our experiments have determined that both the L-Leucine and L-Phenylalanine amino-acid based surfactants exhibit antimicrobial activity, in which the application was stronger against Staphylococcus aureus compared to Escherichia coli.