Chung, Hak Suk Yang, Eun Gyeong Hwang, Dohyeon Lee, Ji Eun Guan, Ziqiang Raetz, Christian R. H. 2024-01-20T09:02:12Z 2024-01-20T09:02:12Z 2021-09-02 2014-09 0006-291X https://pubs.kist.re.kr/handle/201004/126399 The lipopolysaccharide (LPS) isolated from certain important Gram-negative pathogens including a human pathogen Yersinia pestis and opportunistic pathogens Burkholderia mallei and Burkholderia pseudomallei contains D-glycero-o-talo-oct-2-ulosonic acid (Ko), an isosteric analog of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). Kdo 3-hydroxylase (KdoO), a Fe2+/alpha-KG/O-2 dependent dioxygenase from Burkholderia ambifaria and Yersinia pesris is responsible for Ko formation with Kdo(2)-lipid A as a substrate, but in which stage KdoO functions during the LPS biosynthesis has not been established. Here we purify KdoO from B. ambifaria (BaKdoO) to homogeneity for the first time and characterize its substrates. BaKdoO utilizes Kdo(2)-lipid IVA or Kdo(2)-lipid A as a substrate, but not Kdo-lipid IVA in vivo as well as in vitro and Kdo-(Hep)kdo-lipid A in vitro. These data suggest that KdoO is an inner core assembly enzyme that functions after the Kdo-transferase KdtA but before the heptosyl-transferase WaaC enzyme during the Ko-containing LPS biosynthesis. (C) 2014 Elsevier Inc. All rights reserved. English Academic Press Kdo hydroxylase is an inner core assembly enzyme in the Ko-containing lipopolysaccharide biosynthesis Article 10.1016/j.bbrc.2014.08.153 1 Biochemical and Biophysical Research Communications, v.452, no.3, pp.789 - 794 Biochemical and Biophysical Research Communications 452 3 789 794 N scie scopus 000343023900077 2-s2.0-84907511809 Biochemistry & Molecular Biology Biophysics Biochemistry & Molecular Biology Biophysics Article BURKHOLDERIA-CEPACIA COMPLEX ESCHERICHIA-COLI AMINOARABINOSE DIVERSITY MEMBRANE DISACCHARIDE RESISTANCE MUTANT ACID Ko formation Kdo hydroxylase LPS inner core assembly Burkholderia LPS Fe2+/O-2/alpha-KG dependent dioxygenase