Stilbenecarboxylate biosynthesis:: a new function in the family of chalcone synthase-related proteins

被引:67
作者
Eckermann, C
Schröder, G
Eckermann, S
Strack, D
Schmidt, E
Schneider, B
Schröder, J
机构
[1] Univ Freiburg, Inst Biol 2, D-79104 Freiburg, Germany
[2] Inst Biochem Pflanzen, D-06120 Halle An Der Saale, Germany
[3] Max Planck Inst Chem Okol, D-07745 Jena, Germany
关键词
Hydrangea macrophylla; garden hortensia; homology based DNA cloning; chalcone synthase; stilbene synthase; stilbenecarboxylate synthase; recombinant protein expression; hydrangeic acid; phyllodulcin; lunularic acid;
D O I
10.1016/S0031-9422(02)00554-X
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Chalcone (CHS), stilbene (STS) synthases, and related proteins are key enzymes in the biosynthesis of many secondary plant products. Precursor feeding studies and mechanistic rationalization suggest that stilbenecarboxylates might also be synthesized by plant type III polyketide synthases; however, the enzyme activity leading to retention of the carboxyl moiety in a stilbene backbone has not yet been demonstrated. Hydrangea macrophylla L. (Garden Hortensia) contains stilbenecarboxylates (hydrangeic acid and lunularic acid) that are derived from 4-coumaroyl and dihydro-4-coumaroyl starter residues, respectively. We used homology-based techniques to clone CHS-related sequences, and the enzyme functions were investigated with recombinant proteins. Sequences for two proteins were obtained. One was identified as CHS. The other shared 65-70% identity with CHSs and other family members. The purified recombinant protein had stilbenecarboxylate synthase (STCS) activity with dihydro-4-coumaroyl-CoA, but not with 4-coumaroyl-CoA or other substrates. We propose that the enzyme is involved in the biosynthesis of lunularic acid. It is the first example of a STS-type reaction that does not lose the terminal carboxyl group during the ring folding to the end product. Comparisons with CHS, STS, and a pyrone synthase showed that it is the only enzyme exerting a tight control over decarboxylation reactions. The protein contains unusual residues in positions highly conserved in other CHS-related proteins, and mutagenesis studies suggest that they are important for the structure or/and the catalytic activity. The formation of the natural products in vivo requires a reducing step, and we discuss the possibility that the absence of a reductase in the in vitro reactions may be responsible for the failure to obtain stilbenecarboxylates from substrates like 4-coumaroyl-CoA. (C) 2003 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:271 / 286
页数:16
相关论文
共 49 条
[1]   Benzalacetone synthase -: A novel polyketide synthase that plays a crucial role in the biosynthesis of phenylbutanones in Rheum palmatum [J].
Abe, I ;
Takahashi, Y ;
Morita, H ;
Noguchi, H .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 2001, 268 (11) :3354-3359
[2]   THE CONCENTRATIONS OF LUNULARIC ACID AND PRELUNULARIC ACID IN LIVERWORTS [J].
ABE, S ;
OHTA, Y .
PHYTOCHEMISTRY, 1984, 23 (07) :1379-1381
[3]   p-Coumaroyltriacetic acid synthase, a new homologue of chalcone synthase, from Hydrangea macrophylla var. thunbergii [J].
Akiyama, T ;
Shibuya, M ;
Liu, HM ;
Ebizuka, Y .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1999, 263 (03) :834-839
[4]   STUDIES ON PLANT-TISSUE CULTURES .54. NAD(P)H-DEPENDENT 6'-DEOXYCHALCONE SYNTHASE ACTIVITY IN GLYCYRRHIZA-ECHINATA CELLS INDUCED BY YEAST EXTRACT [J].
AYABE, SI ;
UDAGAWA, A ;
FURUYA, T .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1988, 261 (02) :458-462
[5]   ZUR BIOSYNTHESE PFLANZLICHER STILBENE .2. DIE BILDUNG VON RING A DES HYDRANGENOLS [J].
BILLEK, G ;
KINDL, H .
MONATSHEFTE FUR CHEMIE, 1962, 93 (04) :814-&
[6]   Aromatic polyketide synthases - Purification, characterization, and antibody development to benzalacetone synthase from raspberry fruits [J].
BorejszaWysocki, W ;
Hrazdina, G .
PLANT PHYSIOLOGY, 1996, 110 (03) :791-799
[7]   Physics-based modeling and real-time simulation [J].
Chen, JX ;
Yang, YG ;
Wang, XS .
COMPUTING IN SCIENCE & ENGINEERING, 2001, 3 (03) :98-102
[8]  
CROWE J, 1992, QIAEXPRESSIONIST
[9]   6-METHYLSALICYLIC ACID SYNTHETASE FROM PENICILLIUM-PATULUM - SOME CATALYTIC PROPERTIES OF ENZYME AND ITS RELATION TO FATTY-ACID SYNTHETASE [J].
DIMROTH, P ;
RINGELMANN, E ;
LYNEN, F .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1976, 68 (02) :591-596
[10]   Malonyl-CoA:ACP transacylase from Streptomyces coelicolor has two alternative catalytically active nucleophiles [J].
Dreier, J ;
Li, Q ;
Khosla, C .
BIOCHEMISTRY, 2001, 40 (41) :12407-12411