Chalcone and stilbene synthases expressed in eucaryotes exhibit reduced cross-reactivity in vitro

Chalcone synthase (CHS) and stilbene synthase (STS) catalyze different cycl ization reactions of the common tetraketide to give different products, nar ingenin chalcone and resveratrol, respectively. We have previously observed in vitro cross-reaction of CHS and STS overexpressed in Escherichia coli, resveratrol production by CHS and chalcone production by STS. When expresse d in eucaryotic cells, or in E. coli as thioredoxin-fusion proteins, CHS an d STS exhibited reduced cross-reaction. STS refolded from inclusion bodies also showed reduced cross-reaction. While addition of bovine serum albumin and pH in the reaction were without noticeable effect, addition of glycerol decreased the cross-reaction of CHS likely due to its stabilizing effect o n enzyme conformation. These results were interpreted to provide supporting evidence to our earlier proposition (Yamaguchi T. et al., FEBS Lett., 460, 457-461 (1999)) that the in vitro cross-reaction of CHS and STS is due to intrinsic capability of these enzymes to catalyze different types of cycliz ation, which, in turn, is endowed by conformational flexibility of their ac tive sites.