RRC ID 6272
著者 Morikawa T, Saga H, Hashizume H, Ohta D.
タイトル CYP710A genes encoding sterol C22-desaturase in Physcomitrella patens as molecular evidence for the evolutionary conservation of a sterol biosynthetic pathway in plants.
ジャーナル Planta
Abstract We have characterized cytochromes P450, CYP710A13, and CYP710A14, as the sterol C22-desaturase in the moss Physcomitrella patens. GC-MS analyses demonstrated that P. patens accumulated stigmasterol as the major sterol (56-60% of total sterol) and sitosterol to a lesser extent (8-12%); this sterol profile contrasts with those in higher plants accumulating stigmasterol as a minor component. Recombinant CYP710A13 and CYP710A14 proteins prepared using a baculovirus/insect cell system exhibited the C22-desaturase activity with beta-sitosterol to produce stigmasterol, while campesterol and 24-epi-campesterol were not accepted as the substrates. The K(m) values for beta-sitosterol of CYP710A13 (1.0 +/- 0.043 microM) and CYP710A14 (2.1 +/- 0.17 microM) were at comparable levels of those reported with higher plant CYP710A proteins. In Arabidopsis T87 cells over-expressing CYP710A14, stigmasterol contents reached a level 20- to 72-fold higher than those in the basal level of T87 cells, confirming the C22-desaturase activity of this P450 enzyme. The occurrence of the end-products together with the enzymes involved in the last step of the pathway substantiated the presence of an entire sterol biosynthetic pathway in P. patens, providing evidence for the conservation of the sterol biosynthetic pathway through the evolutionary process of land plants.
巻・号 229(6)
ページ 1311-22
公開日 2009-5-1
DOI 10.1007/s00425-009-0916-4
PMID 19306103
MeSH Amino Acid Sequence Animals Biosynthetic Pathways / genetics Bryopsida / enzymology* Bryopsida / genetics Bryopsida / metabolism Cell Line Conserved Sequence / genetics* Cytochrome P-450 Enzyme System / genetics Cytochrome P-450 Enzyme System / metabolism* Electrophoresis, Polyacrylamide Gel Evolution, Molecular Gas Chromatography-Mass Spectrometry Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Isoenzymes / genetics Isoenzymes / metabolism Kinetics Molecular Sequence Data Molecular Structure Oxidoreductases / genetics Oxidoreductases / metabolism Phytosterols / biosynthesis* Phytosterols / chemistry Plants / enzymology* Plants / genetics Plants / metabolism Recombinant Proteins / metabolism Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Sitosterols / chemistry Sitosterols / metabolism Spodoptera Stigmasterol / chemistry Stigmasterol / metabolism
IF 3.39
引用数 19
WOS 分野 PLANT SCIENCES
リソース情報
シロイヌナズナ / 植物培養細胞・遺伝子 rpc00008