TY - JOUR
T1 - Structure function analysis of benzalacetone synthase from Rheum palmatum
AU - Abe, Tsuyoshi
AU - Morita, Hiroyuki
AU - Noma, Hisashi
AU - Kohno, Toshiyuki
AU - Noguchi, Hiroshi
AU - Abe, Ikuro
N1 - Funding Information:
This work was supported by the PRESTO program from Japan Science and Technology Agency, Grant-in-Aid for Scientific Research (Nos. 18510190 and 17310130), and the COE21 program from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
PY - 2007/6/1
Y1 - 2007/6/1
N2 - Benzalacetone synthase (BAS) is a plant-specific chalcone synthase (CHS) superfamily type III polyketide synthase (PKS) that catalyzes a one-step decarboxylative condensation of 4-coumaroyl-CoA with malonyl-CoA. The diketide forming activity of Rheum palmatum BAS is attributed to the characteristic substitution of the conserved active-site Phe215 with Leu (numbering in Medicago sativa CHS). To further understand the structure and function of R. palmatum BAS, four site-directed mutants (C197T, C197G, G256L, and S338V) were newly constructed. All the mutants did not change the product pattern, however, the activity was 2-fold increased in S338V, while reduced to half in G256L mutant. On the other hand, the C197 mutants were functionally almost identical to wild-type BAS, excluding the possibility that the second active-site Cys is involved in the enzyme reaction. Instead, homology modeling suggested a possibility that, unlike the case of CHS, BAS utilizes an alternative pocket to lock the coumaroyl moiety for the diketide formation reaction.
AB - Benzalacetone synthase (BAS) is a plant-specific chalcone synthase (CHS) superfamily type III polyketide synthase (PKS) that catalyzes a one-step decarboxylative condensation of 4-coumaroyl-CoA with malonyl-CoA. The diketide forming activity of Rheum palmatum BAS is attributed to the characteristic substitution of the conserved active-site Phe215 with Leu (numbering in Medicago sativa CHS). To further understand the structure and function of R. palmatum BAS, four site-directed mutants (C197T, C197G, G256L, and S338V) were newly constructed. All the mutants did not change the product pattern, however, the activity was 2-fold increased in S338V, while reduced to half in G256L mutant. On the other hand, the C197 mutants were functionally almost identical to wild-type BAS, excluding the possibility that the second active-site Cys is involved in the enzyme reaction. Instead, homology modeling suggested a possibility that, unlike the case of CHS, BAS utilizes an alternative pocket to lock the coumaroyl moiety for the diketide formation reaction.
KW - Benzalacetone
KW - Benzalacetone synthase
KW - Chalcone synthase
KW - Engineered biosynthesis
KW - Type III polyketide synthase
UR - http://www.scopus.com/inward/record.url?scp=34247863878&partnerID=8YFLogxK
U2 - 10.1016/j.bmcl.2007.03.029
DO - 10.1016/j.bmcl.2007.03.029
M3 - 学術論文
C2 - 17383877
AN - SCOPUS:34247863878
SN - 0960-894X
VL - 17
SP - 3161
EP - 3166
JO - Bioorganic and Medicinal Chemistry Letters
JF - Bioorganic and Medicinal Chemistry Letters
IS - 11
ER -