Bifidobacterium longum requires a fructokinase (Frk; ATP:D-fructose 6-phosphotransferase, EC 2.7.1.4) for fructose catabolism

Journal of Bacteriology
Cristina I CaescuStéphane Bouquelet

Abstract

Although the ability of Bifidobacterium spp. to grow on fructose as a unique carbon source has been demonstrated, the enzyme(s) needed to incorporate fructose into a catabolic pathway has hitherto not been defined. This work demonstrates that intracellular fructose is metabolized via the fructose-6-P phosphoketolase pathway and suggests that a fructokinase (Frk; EC 2.7.1.4) is the enzyme that is necessary and sufficient for the assimilation of fructose into this catabolic route in Bifidobacterium longum. The B. longum A10C fructokinase-encoding gene (frk) was expressed in Escherichia coli from a pET28 vector with an attached N-terminal histidine tag. The expressed enzyme was purified by affinity chromatography on a Co(2+)-based column, and the pH and temperature optima were determined. A biochemical analysis revealed that Frk displays the same affinity for fructose and ATP (Km(fructose) = 0.739 +/- 0.18 mM and Km(ATP) = 0.756 +/- 0.08 mM), is highly specific for D-fructose, and is inhibited by an excess of ATP (>12 mM). It was also found that frk is inducible by fructose and is subject to glucose-mediated repression. Consequently, this work presents the first characterization at the molecular and biochemical level of a fructoki...Continue Reading

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Citations

Dec 11, 2008·Parasitology Research·Kuo-Yang HuangPetrus Tang
Nov 10, 2010·Journal of Proteome Research·Yumiko NakanishiJun Kikuchi
Nov 22, 2011·The Journal of Biological Chemistry·Xiao WeiJing Yuan
Jan 19, 2010·Applied and Environmental Microbiology·Jeong Wook LeeSang Yup Lee
Jan 8, 2014·Bioresource Technology·Min JiangPing Wei
Feb 24, 2015·Parasitology Research·Julia Matt, Michael Duchêne
Mar 22, 2006·Molecular & Cellular Proteomics : MCP·Jing YuanHengliang Wang
Feb 15, 2018·Extremophiles : Life Under Extreme Conditions·Sergey Y ButYuri A Trotsenko
Dec 31, 2014·Applied and Environmental Microbiology·Agnès GroisillierThierry Tonon
Jul 1, 2021·Bioscience, Biotechnology, and Biochemistry·Seiji WatanabeYasuo Ohnishi

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