Chalcone and stilbene synthases expressed in eucaryotes exhibit reduced cross-reactivity in vitro.
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, 457-4 61 (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.
Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Japan.
D Y
DY
Suh
J
J
Kagami
K
K
Fukuma
N
N
Iwanami
Y
Y
Yamazaki
H
H
Yurimoto
Y
Y
Sakai
N
N
Kato
M
M
Shibuya
Y
Y
Ebizuka
U
U
Sankawa
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002175:Candida; D003429:Cross Reactions; D004926:Escherichia coli; D005057:Eukaryotic Cells / Q000201:enzymology / Q000378:metabolism; D002479:Inclusion Bodies / Q000737:chemistry; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D018411:Spodoptera; D013879:Thioredoxins / Q000096:biosynthesis / Q000235:genetics
D011993:Recombinant Fusion Proteins; D013879:Thioredoxins; D000217:Acyltransferases; C022674:stilbene synthase; C020141:flavanone synthetase