The present study was conducted to investigating the effect of Persian gum (PG) and microbial transglutaminase (MTGase) on sensorial, color and microbial characteristics of ultrafiltrated semi-fat white cheese during 60 days of cold storage. In order to produce semi-fat cheeses, PG was used at three levels of 0, 0.25, and 0.5% and MTGase enzyme at three levels of 0, 0.5 and 1 unit/g of protein. The results revealed that the treatment of cheese samples with PG and MTGase enzyme had a positive effect on the sensory and quality characteristics of the product. In general, the cheese sample containing 0.5% PG and 0.5 units of MTGase enzyme attained the highest sensorial scores. Based on panelists’ preference, during the storage time, aroma and texture scores increased while color and appearance attributes decreased. The results obtained from the analysis of color values revealed that the lightness (L*) of cheeses increased with the addition of PG and MTGase enzyme treatment and decreased with the passage of storage time. Unlike the lightness, PG and MTGase enzyme had no significant effect on a* (red-green) and b* (yellow-blue) values of the experimented cheese samples. The results obtained from the microbial evaluation showed that the addition of PG increased the viability of lactic acid bacteria (LAB), but it had no effect on the count of mold and yeasts. On the other hand, increasing the concentration of the enzyme decreased the growth and survival of the studied microorganisms. The results of this study showed that PG can be used as a fat substitute along with MTGase enzyme to produce ultrafiltrated low-fat white cheese with favorable technological and sensory characteristics comparable to high-fat cheese varieties, and the best sample of ultrafiltrated semi-fat cheese is obtained using a treatment containing 0.5% PG and 0.5 unit of MTGase enzyme.