Identification of the glucosamine kinase in the chitinolytic pathway of Thermococcus kodakarensis

Although the chitinolytic pathway of the hyperthermophilic archaeon Thermococcus kodakarensis is well-studied, the genome does not contain genes homologous to previously identified glucosamine kinase genes. As some ADP-dependent glucokinases in the order Thermococcales exhibit phosphorylation activi...

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Bibliographic Details
Published in:Journal of bioscience and bioengineering 2018-03, Vol.125 (3), p.320-326
Main Authors: Aslam, Mehwish, Takahashi, Naoya, Matsubara, Kohei, Imanaka, Tadayuki, Kanai, Tamotsu, Atomi, Haruyuki
Format: Article
Language:English
Subjects:
ADP-dependent glucokinase
Chitinolytic pathway
Glucosamine kinase
Hyperthermophiles
Thermococcus kodakarensis
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Summary:Although the chitinolytic pathway of the hyperthermophilic archaeon Thermococcus kodakarensis is well-studied, the genome does not contain genes homologous to previously identified glucosamine kinase genes. As some ADP-dependent glucokinases in the order Thermococcales exhibit phosphorylation activities for both glucose and glucosamine in vitro, the homolog in T. kodakarensis, encoded by TK1110, was selected as a candidate for the missing glucosamine kinase gene. The purified, recombinant TK1110 enzyme exhibited phosphorylation activities for not only glucose but also glucosamine and N-acetylglucosamine. Kinetic analysis indicated that activity towards glucosamine was as significant as that towards glucose. In order to determine the physiological role of TK1110 in the chitinolytic pathway of T. kodakarensis, a gene disruption strain of TK1110 was constructed. When grown in chitin-containing medium, the TK1110 disruption resulted in almost complete impairment in chitin degradation, and a complete loss of chitin-dependent H2 production. As H2 production is tightly linked to cell growth in T. kodakarensis, the present results strongly suggest that TK1110 functions as the glucosamine kinase responsible for the chitin degradation in T. kodakarensis. [Display omitted] •TK1110 encodes a glucokinase in T. kodakarensis.•The TK1110 protein was proposed to display glucosamine kinase activity.•TK1110 gene deletion resulted in impairment in chitin degradation.•The TK1110 protein functions as both a glucokinase and a glucosamine kinase in T. kodakarensis.
ISSN:1389-1723
1347-4421
DOI:10.1016/j.jbiosc.2017.10.005