Sargassum binderi is a brown algae abundantly available, good nutrient, cheap, does not compete with human, and has not been used optimally as feed for laying hens. Therefore, the purpose of this study was to study the effect of using fermented S. binderi on plasma lipid profiles, fat content, and cholesterol in yolk in laying hens. This study used 200 laying hens (1571-1586 g), aged from 60 to 68 weeks randomized into five different concentrations (0%, 4%, 8%, 12%, and 16%) with a length of 6 weeks. At the end of the study, blood samples were randomly taken from 40 laying hens (2 laying hens per treatment) and 80 eggs were collected randomly (4 eggs per treatment). Variables measured were plasma lipid profile (triglycerides, total cholesterol, and LDL), lipid, and yolk cholesterol. The results showed that the treatment of S. binderi had a significant effect on the lipid profile of laying hens blood serum for total cholesterol and LDL but had no significant impact on blood serum triglycerides. In addition, there was no significant effect on yolk fat and significantly on yolk cholesterol. Thus, the provision of S. binderi to 16% in the laying hen's diet can reduce total blood serum cholesterol from 211.60 to 152.49 mg/dl and LDL from 95.55 to 49.05 mg/dl, with a decrease of 27.93%, and 48.66%, respectively, and decreased yolk cholesterol from 1,279.54 to 1,074.30 mg /100 g with a reduction of 16.04%.