Characterization and Functional Identification of a Gene Encoding Geranylgeranyl Diphosphate Synthase from Dunaliella bardawil.
Geranylgeranyl diphosphate synthase (GGPS) catalyzes the biosynthesis of geranylgeranyl diphosphate, a key precursor for carotenoid biosynthesis. In this study, a full-length cDNA encoding GGPS from Dunaliella bardawil (DbGGPS) was isolated by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of DbGGPS was 1814 bp, containing a 1074 bp ORF encoding 357 amino acids with a calculated mass of 38.88 kDa. Analysis of DbGGPS genomic DNA revealed that it contained 10 exons and 9 introns. It was predicted that DbGGPS possessed a 48 amino acid transit peptide at its N terminus. Bioinformatic analysis revealed that DbGGPS was a member of a group of polyprenyltransferases with five conserved domains and two highly conserved aspartate-rich motifs. Using heterologous expression, carotenoid complementation assay, and gene deletion analysis, it was shown that the coding region of DbGGPS encodes a functional GGPS. This provides new gene sources for carotenoid genetic engineering.
College of Food Science and Engineering, South China University of Technology , Guangzhou 510640, China.
Ming-Hua
MH
Liang
School of Biological Science & Engineering, South China University of Technology , Guangzhou 510006, China.
Ying-Jie
YJ
Liang
College of Food Science and Engineering, South China University of Technology , Guangzhou 510640, China.
Hong-Hao
HH
Jin
College of Food Science and Engineering, South China University of Technology , Guangzhou 510640, China.
Jian-Guo
JG
Jiang
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000460:Chlorophyta / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D051231:Farnesyltranstransferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid
D010940:Plant Proteins; D051231:Farnesyltranstransferase
Dunaliella bardawil; carotenoids; functional expression; geranylgeranyl diphosphate synthase (geranylgeranyl pyrophosphate synthase)