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Kdo hydroxylase is an inner core assembly enzyme in the Ko-containing lipopolysaccharide biosynthesis

Hak Suk Chung1, Eun Gyeong Yang1, Dohyeon Hwang2

  • 1Center for Theragnosis, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea; Department of Biological Chemistry, KIST Campus, Korea University of Science and Technology (UST), Seoul 136-791, Republic of Korea.

Biochemical and Biophysical Research Communications
|September 11, 2014
PubMed

Insights

This study identifies Kdo 3-hydroxylase (KdoO) as an inner core assembly enzyme in lipopolysaccharide (LPS) biosynthesis. It functions after Kdo-transferase and before heptosyl-transferase in Ko-containing LPS production.

Area of Science:

  • Microbiology
  • Biochemistry
  • Glycobiology

Background:

  • Lipopolysaccharide (LPS) is a crucial component of Gram-negative bacteria.
  • Certain pathogens like Yersinia pestis utilize a unique sugar, d-glycero-d-talo-oct-2-ulosonic acid (Ko), in their LPS.
  • The enzyme Kdo 3-hydroxylase (KdoO) is known to be involved in Ko formation, but its precise role in LPS biosynthesis remains unclear.

Purpose of the Study:

  • To purify and characterize KdoO from Burkholderia ambifaria (BaKdoO).
  • To determine the specific substrates and enzymatic activity of BaKdoO.
  • To elucidate the stage at which KdoO functions during Ko-containing LPS biosynthesis.

Main Methods:

  • Purification of BaKdoO to homogeneity.
  • In vitro enzymatic assays using various lipid-linked oligosaccharide substrates.
  • In vivo studies to assess enzyme function within the LPS biosynthesis pathway.

Main Results:

  • BaKdoO was purified and characterized.
  • BaKdoO demonstrated specific utilization of Kdo2-lipid IVA and Kdo2-lipid A as substrates.
  • Substrates like Kdo-lipid IVA and Kdo-(Hep)kdo-lipid A were not utilized by BaKdoO.

Conclusions:

  • KdoO functions as an inner core assembly enzyme in the biosynthesis of Ko-containing LPS.
  • The enzyme acts after Kdo-transferase (KdtA) and before heptosyl-transferase (WaaC).
  • This finding clarifies the sequential order of enzymatic steps in this specific LPS pathway.
Keywords:
Burkholderia LPSFe(2+)/O(2)/α-KG dependent dioxygenaseKdo hydroxylaseKo formationLPS inner core assembly

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