Chalcone and Stilbene Synthases Expressed in Eucaryotes Exhibit Reduced Cross-Reactivity in Vitro

Chem. Pharm. Bull., 48(7), 1051-1054, July 2000

Regular Articles

Chalcone and Stilbene Synthases Expressed in Eucaryotes Exhibit Reduced Cross-Reactivity in Vitro

Dae-Yeon SUH,^a Junichi KAGAMI,^a Kazuki FUKUMA,^a Naoko IWANAMI,^a Yasuyo YAMAZAKI,^a Hiroya YURIMOTO,^b Yasuyoshi SAKAI,^b Nobuo KATO,^b Masaaki SHIBUYA,^c Yutaka EBIZUKA,^c and Ushio SANKAWA^*,a,d

Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University,^a Toyama 930-0194, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University,^b Kyoto 606-8502, Graduate School of Pharmaceutical Sciences, The University of Tokyo,^c Tokyo 113-0033, and International Traditional Medicine Research Center, Toyama International Health Complex,^d Toyama 939-8224, Japan.
Received February 18, 2000; accepted March 24, 2000

Chalcone synthase (CHS) and stilbene synthase (STS) catalyze different cyclization reactions of the common tetraketide to give different products, naringenin 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 expressed in eucaryotic cells, or in E. coli as thioredoxin-fusion proteins, CHS and 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 on 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 cyclization, which, in turn, is endowed by conformational flexibility of their active sites.

Key words Chalcone synthase; stilbene synthase; cross-reaction; heterologous expression

Regular Articles

Chalcone and Stilbene Synthases Expressed in Eucaryotes Exhibit Reduced Cross-Reactivity in Vitro

Dae-Yeon SUH,a Junichi KAGAMI,a Kazuki FUKUMA,a Naoko IWANAMI,a Yasuyo YAMAZAKI,a Hiroya YURIMOTO,b Yasuyoshi SAKAI,b Nobuo KATO,b Masaaki SHIBUYA,c Yutaka EBIZUKA,c and Ushio SANKAWA*,a,d

Dae-Yeon SUH,^a Junichi KAGAMI,^a Kazuki FUKUMA,^a Naoko IWANAMI,^a Yasuyo YAMAZAKI,^a Hiroya YURIMOTO,^b Yasuyoshi SAKAI,^b Nobuo KATO,^b Masaaki SHIBUYA,^c Yutaka EBIZUKA,^c and Ushio SANKAWA^*,a,d