Kinetic Study and Modeling of Wild-Type and Recombinant Broccoli Myrosinase Produced in E. coli and S. cerevisiae as a Function of Substrate Concentration, Temperature, and pH

The myrosinase enzyme hydrolyzes glucosinolates, among which is glucoraphanin, the precursor of anticancer isothiocyanate sulforaphane (SFN). main source glucoraphanin Brassicaceae; however, its natural concentration relatively low, limiting availability SFN. An option to obtain SFN exogenous production, through enzymatic processes and under controlled conditions, allowing complete conversion We characterized kinetics wild-type (BMYR) recombinant broccoli myrosinases produced in E. coli (EMYR) S. cerevisiae (SMYR) terms reaction conditions. Kinetics was adjusted using empirical mechanistic models that describe rate as a function substrate concentration, temperature, pH, resulting R2 values higher than 90%. EMYR differed significantly from those BMYR SMYR probably due absence glycosylations coli. were subjected inhibition but followed different kinetic mechanisms attributed glycosylation patterns. (inactivation Ea = 76.1 kJ/mol) more thermolabile SMYR. showed highest thermostability 52.8 kJ/mol). similar behavior regarding with pK1 (3.4 3.1, respectively) pK2 (5.4 5.0, respectively), considerably