Hemp seed meals contain high protein and unsaturated fatty acids which have the potential as a substitute for soybean meal and yield eggs rich in unsaturated fatty acids. Therefore, the study aimed to evaluate the impact of dietary hemp seed meal as a substitute for soybean meal protein on productive performances, egg quality, and yolk fatty acid composition. A total of 120 Lohmann Brown laying hens aged 50 weeks were separated into 4 groups and 10 repetitions with 3 birds each. Birds received treatment without hemp seed meal (control group), or soybean meal substituted with 4%, 8%, and 12% hemp seed meal. Dietary 4% hemp seed meal significantly increased (p < 0.05) feed efficiency and egg production compared to the 8% and 12% hemp seed meal group. Meanwhile, feed intake, egg weight, body weight loss, egg shape index, albumen index, albumen weight, Haugh unit, yolk weight, yolk index, and eggshell thickness had no significant influence (p > 0.05). However, dietary 8% and 12% hemp seed meal significantly increased (p < 0.05) eggshell weight and yolk color score compared to the control and 4% hemp seed meal group. There was a significant increase (p < 0.05) in omega-3 fatty acid levels and a significant decrease in yolk omega-6 to omega-3 fatty acids ratio with an increase in dietary hemp seed meal. It concluded that dietary 4% hemp seed meal as soybean meal substitution was able to increase egg production, and feed efficiency compared to the level of 8% and 12% hemp seed meal. Increasing dietary levels of hemp seed meal increased eggshell weight, yolk color, and omega-3 fatty acids content, and decreased the omega-6 to omega-3 fatty acids ratio.