Journal of University of Chinese Academy of Sciences >
Cloning and characterization of PcCHS1 from Polygonum cuspidatum
Received date: 2012-03-29
Revised date: 2012-04-13
Online published: 2012-04-13
A full length cDNA encoding a type Ⅲ PKS (PcCHS1) was isolated from Polygonum cuspidatum Sieb. et Zucc. Functional and enzymatic assays biochemically confirmed that PcCHS1 was a chalcone synthase. When incubated with p-coumaroyl-CoA and malonyl-CoA at pH 7-8, PcCHS1 catalyzed the formation of chalcone as the major product. However, at pH 9 both p-hydroxybenzalacetone and chalcone were detected. Site-directed mutagenesis of Phe216 into Leu caused reduction of enzyme activity. Another mutation N333K performed activity of releasing some common derailment products, CTAL and BNY, of many CHSs in vitro.
LI Xing , WANG Hong . Cloning and characterization of PcCHS1 from Polygonum cuspidatum[J]. Journal of University of Chinese Academy of Sciences, 2013 , 30(2) : 206 -212 . DOI: 10.7523/j.issn.1002-1175.2013.02.010
[1] Rawlings B J. Biosynthesis of polyketides (other than actinomycete macrolides)[J]. Natural Product Reports, 1999, 16(4): 425-484.
[2] Whiting D A. Natural phenolic compounds 1900-2000: a bird's eye view of a century's chemistry[J]. Natural Product Reports, 2001, 18(6): 583-606.
[3] Sun Y H, Deng Z X. Polyketides and combinatorial biosynthetic approaches[J]. Chinese Journal of Antibotics, 2006, 31: 6-14 (in Chinese).孙宇辉,邓子新. 聚酮化合物及其组合生物合成[J]. 中国抗生素杂志, 2006, 31: 6-14.
[4] Shen B. Polyketide biosynthesis beyond the type Ⅰ, Ⅱ and Ⅲ polyketide synthase paradigms[J]. Current Opinion in Chemical Biology, 2003, 7(2): 285-295.
[5] Austin M B, Noel J P. The chalcone synthase superfamily of type Ⅲ polyketide synthases[J]. Natural Product Reports, 2003, 20(1): 79-110.
[6] Abe I, Morita H. Structure and function of the chalcone synthase superfamily of plant type Ⅲ polyketide synthases[J]. Natural Product Reports, 2010, 27(6): 809.
[7] Lanz T, Tropf S, Marner F J, et al. The role of cysteines in polyketide synthases-site-directed mutagenesis of resveratrol and chalcone synthases, 2 key enzymes in different plant-specific pathways[J]. Journal of Biological Chemistry, 1991, 266(15): 9971-9976.
[8] Zhang W G, Yang S G, Li B, et al, Advance on chemical ingredients of polygonaceae in China[J]. Progress in Modern Biomedicine, 2008, 8: 393-396 (in Chinese).张文根, 杨赛刚,李波,等. 中国蓼科植物化学成分研究进展[J]. 现代生物医学进展, 2008, 8: 393-396.
[9] Beuerle T, Pichersky E. Enzymatic synthesis and purification of aromatic coenzyme A esters[J]. Analytical Biochemistry, 2002, 302 (2) 305-312.
[10] Ma L, Pang X, Shen H, et al. A novel type Ⅲ polyketide synthase encoded by a three-intron gene from Polygonum cuspidatum[J]. Planta, 2008, 229(3):457-469.
[11] Abe I, Sano Y, Takahashi Y, et al. Site-directed mutagenesis of benzalacetone synthase: the role of PHE215 in plant type Ⅲ polyketide synthases[J]. Journal of Biological Chemistry, 2003, 278(27): 25218-25226.
[12] Abe I, Takahashi Y, Morita H, et al. Benzalacetone synthase: a novel polyketide synthase that plays a crucial role in the biosynthesis of phenylbutanones in Rheum palmatum[J]. Europe Journal of Biochemistry, 2001, 268(11): 3354-3359.
[13] Zheng D, Hrazdina G. Molecular and biochemical characterization of benzalacetone synthase and chalcone synthase genes and their proteins from raspberry (Rubus idaeus L.)[J]. Archives of Biochemistry and Biophysics, 2008, 470(2): 139-145.
[14] Jez J M, Bowman M E, Noel J P. Expanding the biosynthetic repertoire of plant type Ⅲ polyketide synthases by altering starter molecule specificity[J]. Proceedings of the National Academy of Sciences USA, 2002, 99(8): 5319-5324.
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