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Purification and characterization of levoglucosan kinase from Lipomyces starkeyi YZ-215

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A protein named as levoglucosan kinase (EC 2.7.-.-)was purified to homogeneity from a wild isolated strain of Lipomyces starkeyi YZ-215. The protein was purified approximately 30-fold by conventional ammonium sulphate fractionation followed by Resource Q chromatography and two steps of Superdex 200 chromatography, and its physical and kinetic properties were investigated. The purified enzyme showed a molecular weight of 48 kDa by SDS-PAGE and 47.7 kDa by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), respectively. The enzyme was stable at pH 7–10 and showed maximum activity at 30°C and pH 9.0. Kinetic constants (apparent K m values) for levoglucosan and ATP were 68.6 ± 13.7 mM and 0.68 ± 0.06 mM, respectively. After in-gel digestion by trypsin, three peptides were sequenced and analyzed by electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF MS/MS). Data of the amino acid sequences indicated that this protein might be a novel kinase. The purification of levoglucosan kinase from L. starkeyi YZ-215 represented a fundamental step to provide insights into the efficient utilization of cellulosic pyrolysate by bioconversion.

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This work was financially supported by the National Basic Research Program of China (No. 2004CB719704) and National Science Foundation of China (No. 20507021). Dr. Qiangmin Zhang and Dr. Abdul Hamid Khan from the Institute of Microbiology, Chinese Academy of Sciences, were appreciated for their advice and assistance.

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Correspondence to Zhisheng Yu.

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Ning, J., Yu, Z., Xie, H. et al. Purification and characterization of levoglucosan kinase from Lipomyces starkeyi YZ-215. World J Microbiol Biotechnol 24, 15 (2008). https://doi.org/10.1007/s11274-007-9432-5

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  • 1,6-Anhydro-β-d-glucopyranose
  • Levoglucosan kinase
  • Purification
  • Lipomyces starkeyi YZ-215