Development of Saccharomyces cerevisiae Y486–based biosensor carrying both CPR-CYP3A4 and DIN7-GFP constructs for mutagenicity testing of procarcinogens

In the world and Vietnam, a great number of toxic substances from industrial agricultural activities, food production, healthcare services are daily released into environment. Many exogenous harmful procarcinogens, but become carcinogens by bioactivation human cytochrome P450 enzymes (CYPs). Thus, development analytical testing for rapid detection procarcinogens plays crucial role in safety environmental monitoring. This study aims to establish biosensor basing on Saccharomyces cerevisiae Y486 cotransformed with two promoter–gene constructs, CYP3A4–CPR DIN7–GFP. The results showed that all recombinant proteins were coexpressed cells. molecular weight CPR CYP3A4 75 kDa 56 kDa, respectively. enzyme only its catalytic activity biotransformation specific substance as CPR. Kinetic constants, Km, Vmax, Vmax/Km, this CPR–CYP3A4 complex 3.2 µM, 3.5 pmol/pmol CYP/min, 1.1 μL/pmol Coexpressing constructs DIN7-GFP strain was able identify aflatoxin B1 range 0.1 - 0.4 µM; benzo(c)pyrene 10 40 µM. However, system could not detect other procacinogens, such as, N-Nitrosodimethylamine, at any investigated concentrations. These findings first trial further biosensors determine diverse enviroment redesign coexpressing or replacement CYPs inducible promoters.