Biotechnology Letters

, Volume 40, Issue 5, pp 871–880 | Cite as

Identification and characterization of a novel bacterial pyranose 2-oxidase from the lignocellulolytic bacterium Pantoea ananatis Sd-1

  • Keke Zhang
  • Mei Huang
  • Jiangshan Ma
  • Zeyi Liu
  • Jiarui Zeng
  • Xuanming Liu
  • Ting Xu
  • Xiang Wang
  • Ying Liu
  • Zhigang Bu
  • Yonghua Zhu
Original Research Paper
  • 110 Downloads

Abstract

Objective

To identify and characterize a novel bacterial pyranose 2-oxidase (P2Ox) and investigate its potential use in lignin degradation applications.

Results

A new bacterial P2Ox (PaP2Ox) enzyme was identified in the lignocellulolytic bacterium Pantoea ananatis Sd-1. The PaP2Ox open reading frame was cloned, and the encoded protein was heterologously expressed in an Escherichia coli expression system. Unlike another reported bacterial P2Ox enzyme, the purified PaP2Ox exhibits a homotetrameric spatial conformation that is similar to fungal P2Oxs, with each subunit having a molecular mass of 65 kDa. The recombinant PaP2Ox exhibits maximum activity at 50 °C and pH 6.5 with d-glucose as its preferred substrate. In addition, this enzyme was shown to work in combination with bacterial laccase in lignin degradation.

Conclusions

The bacterial enzyme PaP2Ox has potential use in ligninolytic systems and shows promising value in industrial biotechnological applications.

Keywords

Endophytic bacteria Hydrogen peroxide generation Lignin degradation Pantoea ananatis Pyranose 2-oxidase 

Notes

Acknowledgements

This research was supported by the National Natural Science Foundation of China (31672093 and 51378191).

Supporting information

Supplementary Fig. 1—Electrophoretic analysis of PaP2Ox before and after GST tag cleavage.

Supplementary Fig. 2— Fundamental enzymatic characteristics of the cleaved PaP2Ox.

Supplementary material

10529_2018_2538_MOESM1_ESM.tif (19.8 mb)
Supplementary material 1 (TIFF 20315 kb). Supplementary Fig. 1 Electrophoretic analysis of PaP2Ox before and after GST tag cleavage. Lane M, molecular mass marker; lane 1, supernatant of the sonicated extraction from E. coli BL21 (DE3) cells that overexpressed PaP2Ox-containing pGEX-4T-1; lane 2, purified recombinant GST-tagged PaP2Ox; lane 3, the supernatant containing cleaved PaP2Ox protein (65 kDa) and trace amounts of GST-PaP2Ox (91 kDa) and GST tag (26 kDa) after thrombin cleavage; lane 4, the elution mixture containing GST-PaP2Ox (91 kDa), cleaved PaP2Ox (65 kDa) and GST (26 kDa)
10529_2018_2538_MOESM2_ESM.tif (40.5 mb)
Supplementary material 2 (TIFF 41451 kb). Supplementary Fig. 2 Fundamental enzymatic characteristics of the cleaved PaP2Ox. The optimum temperature (a) of PaP2Ox was measured at pH 6.5. The thermostability (b) of PaP2Ox was analysed at pH 6.5 for 30 min, and the residual activity was measured under the standard assay conditions. The optimum pH (c) of PaP2Ox was measured at room temperature. The pH (d) stability of PaP2Ox was investigated at 4°C, and the residual activity was measured under the standard assay conditions

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Keke Zhang
    • 1
  • Mei Huang
    • 1
  • Jiangshan Ma
    • 2
  • Zeyi Liu
    • 1
  • Jiarui Zeng
    • 1
  • Xuanming Liu
    • 1
  • Ting Xu
    • 1
  • Xiang Wang
    • 1
  • Ying Liu
    • 1
  • Zhigang Bu
    • 1
  • Yonghua Zhu
    • 1
  1. 1.Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, College of BiologyHunan UniversityChangshaPeople’s Republic of China
  2. 2.Hunan Academy of ForestryChangshaPeople’s Republic of China

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