Instant noodles gained widespread popularity, resulting in a surge in global wheat consumption. However, concerns about the health implications of wheat-based products spurred the need for alternative ingredients in noodle production. The substitution of wheat flour with local resources, such as mocaf flour, and the enrichment of noodles with latoh (Caulerpa lentillifera), provided an opportunity to reduce wheat imports while enhancing the quality and potential health benefits of noodles. This study aimed to determine the optimal formulation, assess the characteristics, and evaluate the functional properties of wheat noodles that were substituted with mocaf flour and supplemented with latoh. The formulation of these noodles was conducted using design expert. The research encompassed a comprehensive analysis of physical, chemical, and organoleptic properties. The presence of latoh, in terms of chemical characteristics, increased the water and protein content in the noodles. On the physical side, it increased the final viscosity, setback, peak time, and elongation, as well as reduced the peak viscosity and breakdown. Meanwhile, the presence of mocaf increased carbohydrate content, peak viscosity, and breakdown value. The variations in noodle formulas only affected consumer acceptance of the appearance of the noodles; the noodle formula 99:1 (Mocaf:Latoh) had the most preferred appearance. Based on the sensory test parameters, the elongation value, and the tensile strength, noodles with optimal formulas were obtained, namely mocaf 63%, wheat 36% and latoh 1%. The optimum formula was similar to sample 10 (60:39:1) which exhibited characteristics such as a bright color, a rather smooth appearance with no cracks, a very soft mouthfeel, a bit springiness and elasticity, and a slightly non-sticky texture to the teeth without a fishy aroma. The essential and non-essential amino acids with the highest concentration contained in the Mocaf Noodles’ optimum formula were glutamate and leucine. Mocaf noodles with added latoh had low fat content but high carbohydrates and were rich in essential amino acids compared to other noodles. Additionally, they possessed sensory characteristics that were deemed acceptable by the panelists.