Optimization of microbial transglutaminase production using experimental designs

In prokaryotes, transglutaminase (TGase) has been found only in actinomycetes from the genus Streptoverticillium. The role of this TGase, as well as the mechanism regulating the enzyme expression, are still unknown. In order to improve TGase production by Streptoverticillium cinnamoneum CBS 683.68 and simultaneously elucidate the relationship between growth and TGase activity, we decided to study these two responses using different designs of statistical analysis. Among the five factors tested? casein, glycerol, peptones, yeast extract and oligoelements, only oligoelements were found to have no effect either on growth or on TGase production in a complete factorial design. The two factors casein and glycerol were found to have a highly significant effect on both dry weights and TGase activity in a Box-Behnken design used to improve the model. Finally, the TGase activity was increased three times to reach 0.331 +/- 0.038 U/ml with optimum concentrations of casein (38.4 g/l) and glycerol (31.2 g/l) calculated with the help of a composite design. In the course of these experiments, the two responses varied in the same way, demonstrating that growth and TGase production were tightly correlated under the conditions described. However, TGase was produced during the stationary phase of growth in optimized medium, indicating that the enzyme production could be induced.