Fenugreek is one of the spice that have an antibacterial compound. Pepper and salt have also been used as combination in this marinade and have antibacterial compound. This research was aimed to study the antimicrobial effect of marinade base on fenugreek as natural preservative agent toward tofu in the two different temperatures storage, which were room storage (≈24°C) and cold storage (≈5°C). The fenugreek base marinade was tested against three bacterias, Escherichia coli, Staphylococcus aureus and Bacillus cereus by the well diffusion method. From the statistical result, the marinade with 6%, 8% and 10% concentration of fenugreek gave the significantly different in the result. The main research used three different concentrations of fenugreek, which were 6%, 8%, and 10). The fenugreek base marinate with those concentration results a non toxic. Active compounds as alkaloid, saponin, tannin, flavonoid, triterphenoid, and glycoside are determined by phytochemical test. Those test showed that 10% concentration of the fenugreek on marinade is the best concentration of all. Microbiology test showed that marinade base on fenugreek can preserve tofu up to 1 days on room storage and 5 days in cold storage. Challenge test result showed that fenugreek base marinate with 10% concentration was the best that can inhibit bacterial growth and the most accepted by panelists.