Transglutaminase catalyzes the cross-linking reaction between a glutamine residue and a free amine residue of proteins leading to the formation of protein aggregates. In this research, the effects of temperature, agitation, and aeration on the production of transglutaminase in a bench reactor by a newly isolated Streptomyces sp. from Brazilian soils were investigated using a factorial experimental design. The parameters evaluated influenced the enzyme production, and the data showed that the best conditions to enhance cell growth were different from those leading to enhanced transglutaminase production. Thus, a temperature and agitation shift strategy was adopted to increase transglutaminase productivity. The temperature and agitation were first set at 34 A degrees C and 350 rpm, respectively, and after 24 h decreasing to 26 A degrees C and 150 rpm until the end of fermentation. The transglutaminase activity obtained was 2.18 U/mL after 42 h of fermentation, which was twice than that obtained using a constant temperature and agitation fermentation strategy.