title: Molecular cloning and characterization of a flavanone 3-Hydroxylase gene from Artemisia annua L.
abstract: Flavonoids were found to synergize anti-malaria and anti-cancer compounds in Artemisia annua, a very important economic crop in China. In order to discover the regulation mechanism of flavonoids in Artemisia annua, the full length cDNA of flavanone 3-hydroxylase (F3H) were isolated from Artemisia annua for the first time by using RACE (rapid amplification of cDNA ends). The completed open read frame of AaF3H was 1095 bp and it encoded a 364-amino acid protein with a predicted molecular mass of 41.18 kDa and a pI of 5.67. The recombinant protein of AaF3H was expressed in E. coli BL21(DE3) as His-tagged protein, purified by Ni-NTA agrose affinity chromatography, and functionally characterized in vitro. The results showed that the His-tagged protein (AaF3H) catalyzed naringenin to dihydrokaempferol in the present of Fe(2+). The Km for naringenin was 218.03 μM. The optimum pH for AaF3H reaction was determined to be pH 8.5, and the optimum temperature was determined to be 35 °C. The AaF3H transcripts were found to be accumulated in the cultivar with higher level of flavonoids than that with lower level of flavonoids, which implied that AaF3H was a potential target for regulation of flavonoids biosynthesis in Artemisia annua through metabolic engineering.
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Xiong
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China; National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals, China.
authors.forename: Na
authors.initials: N
authors.lastname: Tian
authors.affiliation: National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals, China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Long
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Jinyu
authors.initials: J
authors.lastname: Feng
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China; National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals, China. Electronic address: xiaowenjun88@sina.com.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Xiao
authors.affiliation: Hunan Collaborative Innovation for Utilization of Botanical Functional Ingredients, College of Horticulture and Hardening, Hunan Agricultural University, Changsha, 410128, China; National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals, China; Key Lab of Tea Science, Ministry of Education, Changsha, 410128, China. Electronic address: shqianliu@sina.com.
authors.forename: Shuoqian
authors.initials: S
authors.lastname: Liu
mesh_terms: D000872:Anthocyanins / Q000494:pharmacology; D031172:Artemisia annua / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D044950:Flavanones / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D056726:Genetic Association Studies; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics*; D010802:Phylogeny; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity / Q000187:drug effects
chemical_list: D000872:Anthocyanins; D044950:Flavanones; D010940:Plant Proteins; D011994:Recombinant Proteins; D006899:Mixed Function Oxygenases; C048797:flavanone 3-dioxygenase; C005273:naringenin
keywords: Artemisia annua L.; Dihydrokaempferol; Flavanone 3-hydroxylase; Flavonoids; Naringenin