Application of numerical modeling for the development of optimized complex medium for D-hydantoinase production from...

D-Hydantoinases (E.C.3.5.2.2) are commercially valuable enzymes involved in the production of D-amino acids. However, commercial exploitation of the biological process is rare, mainly because sufficient details are not available on the efficient production of these enzymes by microorganisms. In the present study, Agrobacterium radiobacter was used as the source of D-hydantoinase and its production was optimized with inexpensive carbon and nitrogen sources. The four media components selected to study their effect on biomass and/or enzyme activities were molasses, ammonium nitrate, sodium di-hydrogen orthophosphate, and manganese chloride. With the use of an empirical modeling technique (response surface method), we have optimized both biomass and enzyme production in this organism, with a minimal number of batches. Experiments were performed with optimized media components to validate the model. The maximum level of enzyme and biomass obtained was 35 U/mL and 1.69 mg/mL, respectively.