The effect of spraying microbial transglutaminase (MTGase) at various levels (1, 4, 7, and 10 units/gram) on the quality of mackerel (Scomber scombrus) fillets was investigated during refrigerated storage. The results of pH and the total volatile base nitrogen (TVB-N) revealed that 7 mu/g MTGase treatment met the best results regarding fish quality. The oxidation was hindered by increasing MTGase concentration, and the lowest totox value was recorded in the fillets treated with 10 mu MTGase. The progress in the total free amino acids was suppressed with the addition of MTGase, and the most effective concentration was found to be 7 mu/g MTGase treatment. Moreover, this concentration was the most promising in inhibition of the total mesophilic aerobic bacteria, the total psychrophilic aerobic bacteria, coliform, and yeast-mold. Panelists preferred the samples treated with 7 mu/g MTGase in terms of odor, texture, and appearance. The addition of MTGase straightened the fish muscle and helped preserve its initial texture. It is important to use the proper concentration of MTGase instead of high concentrations.