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Cloning and Characterization of D-xylulose Kinase from Kocuria gwangalliensis Strain SJ2
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  • Journal title : Journal of Life Science
  • Volume 25, Issue 5,  2015, pp.507-514
  • Publisher : Korean Society of Life Science
  • DOI : 10.5352/JLS.2015.25.5.507
 Title & Authors
Cloning and Characterization of D-xylulose Kinase from Kocuria gwangalliensis Strain SJ2
Jeong, Tae Hyug; Hwang, Tae Kyung; Seo, Yong Bae; Kim, Young Tae;
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D-Xylulose is phosphorylated to D-xylulose-5-phosphate by D-xylulose kinase before it enters glycolysis via the nonoxidative pentose phosphate pathway. A gene encoding a novel D-xylulose kinase (XK) from K. gwangalliensis strain SJ2 was sequenced and expressed in E. coli. The sequence of the isolated XK gene was 1,419 bp, encoding 472 amino acids. The XK protein was more closely related to the Arthrobacter phenanthrenivorans XK than to the Bifidobacterium catenulatum one, as reflected in the sequence identity (54.9% vs. 38.7%). The XK gene was subcloned into the pCold-II expression vector. The resulting plasmid was transformed into E. coli strain BL21 (DE3) cells and the expression of the recombinant XK protein was induced by the addition of IPTG. The resulting protein was expressed as a fusion protein of approximately 48 kDa containing a N-terminal six-histidine extension that was derived from the expression vector. The expressed protein was homogenized by affinity chromatography and showed enzymatic activity corresponding to D-xylulose kinase. XK enzyme kinetic studies with D-xylulose and ATP showed a Km of 250±20 μM and 1,300±50 μM, respectively. The results obtained from this study will provide a wider knowledge base for the characterization of D-xylulose kinase at the molecular level.
Characterization;D-xylulose kinase;enzyme kinetic;gene cloning;Kocuria gwangalliensis;
 Cited by
Deng, X. X. and Ho, N. W. Y. 1990. Xylulokinase Activity in various yeasts including Saccharomyces cerevisiae containing the cloned Xylulokinase gene. Appl. Biochem. Biotech. 24, 193-199.

Flanagan, T. and Waites, M. J. 1992. Purification and characterization of D-xylulokinase from the pentose-fermenting yeast Pichia stipidis NCYC 1541. Enzyme Microb. Technol. 14, 975-979. crossref(new window)

Ho, N. W. Y. and Chang, S. F. 1989. Cloning of yeast Xylulokinase gene by complementation of E. coli and yeast mutations. Enzyme Microb. Technol. 11, 417-421. crossref(new window)

Johansson, B., Christensson, C., Hobley, T. and Hahn-Ha¨erdal, B. 2001. Xylulokinase overexpression in two strains of Saccharomyces cerevisiae also expressing xylose reductase and xylitol dehydrogenase and its effect on fermentation of xylose and lignocellulosic hydrolysate. Appl. Environ. Microbiol. 67, 4249-4255. crossref(new window)

Lawlis, V. B., Dennis, M. S., Chen, E. Y., Smith, D. H. and Henner, D. J. 1984. Cloning and Sequencing of the Xylose Isomerase and Xylulose Kinase genes of Escherichia coli. Appl. Environ. Microbiol. 47, 15-21.

Lee, T. H., Kim, M. D. and Park, Y. C. 2003. Effects of xylulokinase activity on ethanol production from D-xylulose by recombinant Saccharomyces cerevisiae. J. Appl. Microbiol. 95, 847-852. crossref(new window)

Richard, P., Toivari, M. H. and Penttila, M. 2000. The role of xylulokinase in Saccaromyces cerevisiae xylulose catabolism. FEMS Microbiol. Lett. 190, 39-43. crossref(new window)

Rodriguez-Pena, J. M., Cid, V. J., Arroyo, J. and Nombela, C. 1998. The YGR194c (XKS1) gene encodes the Xylulokinase from the budding yeast Saccharomyces cerevisiae. FEMS Microbiol. Lett. 162, 155-160. crossref(new window)

Rosenfeld, S. A., Stevis, P. E. and Ho, N. W. 1984. Cloning and Characterization of the xyl genes from Escherichia coli. Mol. Gen. Genet. 194, 410-415. crossref(new window)

Seo, Y. B., Kim, D. E., Kim, G. D., Kim, H. W., Nam, S. W., Kim, Y. T. and Lee, J. H. 2009. Kocuria gwangalliensis sp. nov., an Actinobacterium isolated from Seawater. Int. J. Syst. Evol. Microbiol. 59, 2769-2772. crossref(new window)

Jeong, T. H., Ji, K. H. and Kim, Y. T. 2013. Overexpression and characterization of lycopene cyclase (CrtY) from marine bacterium, Paracoccus haeundaensis. J. Microbiol. Biotechnol. 23, 144-148. crossref(new window)

Jeong, T. H., Youn, J. Y., Ji, K. H., Seo, Y. B. and Kim, Y. T. 2014. Cloning and characterization of phosphoinositide 3-Kinase γ cDNA from flounder (Paralichthys olivaceus). J. Life Sci. 24, 343-351. crossref(new window)

Toivari, M. H., Aristidou, A., Ruohonen, L. and Penttila, M. 2001. Conversion of xylose to ethanol by recombinant Saccharomyces cerevisiae: importance of xylulokinase (XKS1) and oxygen availability. Metab. Eng. 3, 236-249. crossref(new window)