[Oxidase gene from sweetpotato].
Polyphenol oxidase is the enzyme responsible for enzymatic browning in sweetpotato that decreases the commercial value of sweetpotato products. Here we reported the cloning and characterization of a new cDNA encoding PPO from sweetpotato, designated as IbPPO (GeneBank accession number: AY822711). The full-length cDNA of IbPPO is 1984 bp with a 1767 bp open reading frame (ORF) encoding a 588 amino acid polypeptide with calculated molecular weight of 65.7 kDa and theoretical pI of 6.28. The coding sequence of IbPPO was also directly amplified from the genomic DNA of sweetpotato that demonstrated that IbPPO was an intron-free gene. The computational comparative analysis revealed that IbPPO showed homology to other PPOs of plant origin and contained a 50 amino acid plastidial transit peptides at its N-terminal and the two conserved CuA and CuB copper-binding motifs in the catalytic region of IbPPO. A highly conserved serine-rich motif was firstly found in the transit peptides of plant PPO enzymes. Then the homology-based structural modeling of IbPPO showed that IbPPO had the typical structure of PPO: the catalytic copper center was accommodated in a central four-helix bundle located in a hydrophobic pocket close to the surface. Finally, the results of the semi-quantitative RT-PCR analysis of IbPPO in different tissues demonstrated that IbPPO could express in all the organs of sweetpotato including: mature leaves, young leaves, the stems of mature leaves (petioles), the storage roots and the veins but at different levels. The highest-level expression of IbPPO was found in veins, followed by storage roots, young leaves and mature leaves; and the lowest-level expression of IbPPO was found in petioles. The present researches will facilitate the development of anti-brown sweetpotato by genetic engineering.
Liao
Zhihua
Z
Chen
Rong
R
Chen
Min
M
Yang
Yijian
Y
Fu
Yufan
Y
Zhang
Qitang
Q
Lan
Xiaozhong
X
D020816:Amino Acid Motifs; D001483:Base Sequence; D004156:Catechol Oxidase / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D027723:Ipomoea batatas / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D011487:Protein Conformation
D018076:DNA, Complementary; D004156:Catechol Oxidase