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Identification of 2-keto-3-deoxy-D-Gluconate Kinase and 2-keto-3-deoxy-D-Phosphogluconate Aldolase in an Alginate-Assimilating Bacterium, Flavobacterium sp Strain UMI-01

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Title: Identification of 2-keto-3-deoxy-D-Gluconate Kinase and 2-keto-3-deoxy-D-Phosphogluconate Aldolase in an Alginate-Assimilating Bacterium, Flavobacterium sp Strain UMI-01
Authors: Nishiyama, Ryuji Browse this author
Inoue, Akira Browse this author →KAKEN DB
Ojima, Takao Browse this author →KAKEN DB
Keywords: alginate degradation
4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH) metabolism
2-keto-3-deoxy-D-gluconate (KDG) kinase
2-keto-3deoxy-6-phosphogluconate (KDPG) aldolase
alginate-derived products
Issue Date: Feb-2017
Publisher: MDPI
Journal Title: Marine drugs
Volume: 15
Issue: 2
Start Page: 37
Publisher DOI: 10.3390/md15020037
Abstract: Recently, we identified an alginate-assimilating gene cluster in the genome of Flavobacterium sp. strain UMI-01, a member of Bacteroidetes. Alginate lyase genes and a 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH) reductase gene in the cluster have already been characterized; however, 2-keto-3-deoxy-D-gluconate (KDG) kinase and 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase genes, i.e., flkin and flald, still remained uncharacterized. The amino acid sequences deduced from flkin and flald showed low identities with those of corresponding enzymes of Saccharophagus degradans 2-40T, a member of Proteobacteria (Kim et al., Process Biochem., 2016). This led us to consider that the DEH-assimilating enzymes of Bacteroidetes species are somewhat deviated from those of Proteobacteria species. Thus, in the present study, we first assessed the characteristics in the primary structures of KDG kinase and KDG aldolase of the strain UMI-01, and then investigated the enzymatic properties of recombinant enzymes, recFlKin and recFlAld, expressed by an Escherichia coli expression system. Multiple-sequence alignment among KDG kinases and KDG aldolases from several Proteobacteria and Bacteroidetes species indicated that the strain UMI-01 enzymes showed considerably low sequence identities (15%-25%) with the Proteobacteria enzymes, while they showed relatively high identities (47%-68%) with the Bacteroidetes enzymes. Phylogenetic analyses for these enzymes indicated the distant relationship between the Proteobacteria enzymes and the Bacteroidetes enzymes, i.e., they formed distinct clusters in the phylogenetic tree. recFlKin and recFlAld produced with the genes flkin and flald, respectively, were confirmed to show KDG kinase and KDPG aldolase activities. Namely, recFlKin produced 1.7 mM KDPG in a reaction mixture containing 2.5 mM KDG and 2.5 mM ATP in a 90-min reaction, while recFlAld produced 1.2 mM pyruvate in the reaction mixture containing 5 mM KDPG at the equilibrium state. An in vitro alginate-metabolizing system constructed from recFlKin, recFlAld, and previously reported alginate lyases and DEH reductase of the strain UMI-01 could convert alginate to pyruvate and glyceraldehyde-3-phosphate with an efficiency of 38%.
Type: article
Appears in Collections:水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 尾島 孝男

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