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Cloning and heterologous expression of isopentenyl diphosphate isomerase gene from Lycium chinense

  • Zhaodi Li
  • Jing JiEmail author
  • Gang Wang
  • Tchouopou Lontchi Josine
  • Jiang Wu
  • Jinjin Diao
  • Weidang Wu
  • Chunfeng Guan
Original Article

Abstract

Isopentenyl diphosphate isomerase (IPI), a key enzyme of terpenoid biosynthesis pathway, catalyzes the reversible conversion between isopentenyl diphosphate (IPP) and its allylic isomer dimethylallyl diphosphate (DMAPP), which are essential precursors for terpenoid biosynthesis. In this study, we isolated and characterized the IPI gene from Lycium chinense (LcIPI). The full-length cDNA sequence was 1104 bp encoding 293 amino acid residues. Additionally, several specific conserved motifs of IPI were also identified by amino acid sequence analysis in this protein. Phylogenetic tree showed that LcIPI had a close relationship with other solanaceae plants. Expression of recombinant protein in Escherichia coli produced a clear target band of about 35 kDa. Enzymatic activity was determined by heterologous complementation analysis, and β-carotene content of LcIPI transformants was about two times more compared with control. Furthermore, quantitative real-time PCR assay demonstrated that LcIPI was expressed in all tissues tested with the highest level in flowers. The results obtained here clearly suggest that the LcIPI gene is a promising candidate for carotenoid production in heterologous hosts.

Keywords

Lycium chinense Isopentenyl diphosphate isomerase Heterologous expression Carotenoid biosynthesis 

Abbreviations

IPP

Isopentenyl diphosphate

DMAPP

Dimethylallyl diphosphate

IPI

Isopentenyl diphosphate isomerase

HPLC

High-performance liquid chromatography

Notes

Acknowledgments

The authors appreciate the kind presentation of plasmid pACCAR16ΔcrtX from professor Chanfu Zhu (School of Life Sciences, Northeast Normal University, Changchun, China). This work was financially supported by the National Science and Technology Key Project of China on GMO Cultivation for New Varieties (No. 2014ZX08003-002B).

Conflict of interest

The authors declare that they do not have any conflict of interest.

Supplementary material

13562_2015_306_MOESM1_ESM.doc (651 kb)
Supplementary 1 Partial sequence of isopentenyl diphosphate isomerase gene from Lycium chinense. (DOC 651 kb)
13562_2015_306_MOESM2_ESM.doc (4 mb)
Supplementary 2 Three-dimensional model of the Lycium chinense IPI. The ribbons and arrows stand for α-helix and β-sheet regions, respectively. The loop regions are also present in the schematics. (DOC 4108 kb)
13562_2015_306_MOESM3_ESM.doc (37 kb)
Supplementary 3 Primers sequence of Arabidopsis thaliana and Zea mays. Underline stands for the restriction site of BamHI and XholI respectively. (DOC 37 kb)

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Copyright information

© Society for Plant Biochemistry and Biotechnology 2015

Authors and Affiliations

  • Zhaodi Li
    • 1
  • Jing Ji
    • 2
    Email author
  • Gang Wang
    • 2
  • Tchouopou Lontchi Josine
    • 1
  • Jiang Wu
    • 1
  • Jinjin Diao
    • 1
  • Weidang Wu
    • 1
  • Chunfeng Guan
    • 2
  1. 1.School of Chemical Engineering and TechnologyTianjin UniversityTianjinPeople’s Republic of China
  2. 2.School of Environmental Science and EngineeringTianjin UniversityTianjinPeople’s Republic of China

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