Plastidial α-glucan phosphorylase 1 complexes with disproportionating enzyme 1 in Ipomoea batatas storage roots for elevating malto-oligosaccharide metabolism

PLoS One. 2017 May 4;12(5):e0177115. doi: 10.1371/journal.pone.0177115. eCollection 2017.

Abstract

It has been proposed that malto-oligosaccharides (MOSs) are possibly recycled back into amylopectin biosynthesis via the sequential reactions catalyzed by plastidial α-glucan phosphorylase 1 (Pho1) and disproportionating enzyme 1 (Dpe1). In the present study, the reciprocal co-immunoprecipitation experiments using specific antibodies against Pho1 and Dpe1 demonstrated that these two enzymes can form a complex (the PD complex) in Ipomoea batatas storage roots. The immunohistochemistry analyses also revealed the co-localization of Pho1 and Dpe1 in the amyloplasts, and the protein levels of Pho1 and Dpe1 increased gradually throughout sweet potato storage root development. A high molecular weight PD complex was co-purified from sweet potato storage root lysates by size exclusion chromatography. Enzyme kinetic analyses showed that the PD complex can catalyze maltotriose and maltotetraose to generate glucose-1-phosphate in the presence of inorganic phosphate, and it also performs greater Dpe1 activity toward MOSs than does free form Dpe1. These data suggest that Pho1 and Dpe1 may form a metabolon complex, which provides elevated metabolic fluxes for MOS metabolism via a direct transfer of sugar intermediates, resulting in recycling of glucosyl units back into amylopectin biosynthesis more efficiently.

MeSH terms

  • Catalysis
  • Chromatography, Gel
  • Immunoprecipitation
  • Ipomoea / enzymology*
  • Ipomoea / metabolism
  • Mass Spectrometry
  • Oligosaccharides / metabolism*
  • Phosphorylases / metabolism*
  • Plant Roots / enzymology*
  • Plant Roots / metabolism
  • Plastids / enzymology*

Substances

  • Oligosaccharides
  • Phosphorylases

Grants and funding

This study was supported by the Ministry of Science and Technology, Taiwan (105-2119-M-001-014 awarded to SCC; 103-2313-B-002-030-MY3 awarded to RHJ), https://www.most.gov.tw/. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.