Turkey ranks third in the world in terms of tomato production, and approximately 12.5 million tons of tomatoes are produced annually. Especially in greenhouse cultivation, tomato wastes do not stay in the soil and are taken out of the greenhouse. Ten million tons of tomato waste is generated annually in Turkey. Compost is a very important organic material containing plant nutrients such as nitrogen, phosphorus, potassium, magnesium, and microelements. Tomato waste compost is a good organic fertilizer that increases both tomato yield and biological activities in soil. The aim of this study is to determine the effect of tomato compost (TC) on tomato yield and biological properties of soils. For this purpose, a pot experiment was established in the greenhouse and TC was applied in four different doses (TC1—1%, TC2—2%, TC3—4%, TC4—6%) to pots containing 3.5 kg of soil, these doses were also applied with chemical fertilizer (CF) (CF, TC1 + CF, TC + CF, TC3 + CF, TC4 + CF), and then tomato plants were grown. Each application was applied as three replications. Among the compost applications, the highest tomato yield was obtained with the TC4 (0.96 kg/pot, 14.4 t/ha) application: 15.66% higher yield was obtained compared to chemical fertilizers. Among all treatments, TC1 + CF (1.07 kg/pot, 16.10 t/ha), which increased the tomato yield by 28.9%, had the highest yield. All applications have increased CO2 production in the soil; the highest CO2 production was determined in the last harvest period with TC3 + CF and TC4 + CF (0.27 mgCO2 1 gr−1 24 h−1). The maximum microbial biomass carbon content was determined in the first harvest period. Among the applications, the most microbial biomass carbon was determined after the TC4 + CF (114.42 mgMBC 100 gr−1) application. Catalase enzyme activity was obtained the most with TC4 + CF (601.56 mlO2 1 gr−1) application in first florescence. Dehydrogenase enzyme activity was obtained the most with TC3 (55.96 μg TPF g−1) application in end of harvest. After compost application, tomato yield increased and biological properties of soil improved.