title: Expression and Enzyme Activity Detection of a Sepiapterin Reductase Gene from Musca domestica Larva.
abstract: Tetrahydrobiopterin (BH4) is an essential cofactor for aromatic acid hydroxylases and nitric oxide synthase. Sepiapterin reductase (SPR) catalyzes the final steps of BH4 biosynthesis. Studies on SPR from several insects and other organisms have been reported. However, thus far, enzyme activity of SPR in Musca domestica is kept unknown. In this study, 186 differentially expressed genes including SPR gene from Musca domestica (MDSPR) were screened in subtractive cDNA library. The MDSPR gene was cloned, and the recombinant MDSPI16 protein was expressed as a 51-kDa protein in soluble form. The MDSPR exhibited strong activity to the substrate sepiapterin (SP). The values of Vmaxand Kmof the MDSPR for SP were 6.83 μM/min and 23.48 μM, and the optimum temperature and pH of MDSPR were 50 °C and 4.0, respectively. This study provides new hypotheses and methods for the production of BH4 using insect-derived SPR.
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Tang
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Pei
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.|Jilin Medical University, Jilin Street No.5, Jilin, Jilin Province, 132013, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.affiliation: Chengdu tongwei Co. Ltd,houwu district, Chengdu, Sichuan Province, 610041, China.
authors.forename: Dongfang
authors.initials: D
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.
authors.forename: Lingcong
authors.initials: L
authors.lastname: Kong
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.
authors.forename: Shuming
authors.initials: S
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Jiang
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China.
authors.forename: Yunhang
authors.initials: Y
authors.lastname: Gao
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Xincheng Street No.2888, Changchun, 130118, China. hongxia0731001@163.com.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Ma
mesh_terms: D000429:Alcohol Oxidoreductases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D001708:Biopterins / Q000031:analogs & derivatives* / Q000096:biosynthesis; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006793:Houseflies / Q000201:enzymology* / Q000235:genetics; D007814:Larva / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data
chemical_list: D001708:Biopterins; D000429:Alcohol Oxidoreductases; C022630:sepiapterin reductase; C003402:sapropterin
keywords: Enzyme activity; Gene cloning; Musca domestica; Protein expression; Sepiapterin reductase