Flavin-Based Fluorescent Protein EcFbFP Auto-Guided Surface Display of Methyl Parathion Hydrolase in Escherichia coli

  • Lu Bian
  • Zhen Zhang
  • Rong-xing Tang
  • Wei Shen
  • Li-xin MaEmail author
Original paper


Methyl parathion hydrolase (MPH) plays an important role in degrading a range of organophosphorus compounds. In order to display MPH on the cell surface of Escherichia coli strain RosettaBlue™, the Flavin-based fluorescent protein EcFbFP was severed as an auto-anchoring matrix. With net negative charges of EcFbFP supplying the driving forces, fusion protein MPH-EcFbFP through a two-step auto-surface display process was finally verified by (a) inner membrane translocation and (b) anchoring at outer membrane. Cells with surface-displayed MPH obtained activity of 0.12 U/OD600 against substrate methyl parathion. MPH when fused with engineered EcFbFP containing 20 net negative charges exhibited fivefold higher anchoring efficiency and tenfold higher enzymatic catalytic activity of 1.10 U/OD600. The above result showed that MPH was successfully displayed on cell surface and can be used for biodegradation of methyl parathion.


Flavin-based fluorescent protein EcFbFP Cell-surface display Net negative charge Methyl parathion hydrolase 



Flavin mononucleotide


Methyl parathion hydrolase


Escherichia coli codon-optimized FMN-based fluorescent protein


EcFbFP mutant with 0 net charge


EcFbFP mutant with 2 net positive charges


EcFbFP mutant with 4 net positive charges


EcFbFP mutant with 20 net negative charges


MPH fused with EcFbFP


MPH fused with EcFbFP(-20)


Super folder green fluorescent protein


MCherry fluorescent protein


MPH fused with sfGFP


MPH fused with mCherry



The authors would like to thank professor Li Yi in Hubei University for the critical revision of the manuscript, thank Dr. Mohanty P.B. in University of Oklahoma (U.S.A.) for the analysis of protein structure.

Author Contributions

L.B., Z.Z., and L.M. conceived, designed, and coordinated the study, and wrote the paper. R.T. performed the experiments shown in Figs. 1, 2, 3, 4, and 5. L.B. performed tables, and contributed to paper writing. W.S. performed the experiments shown in Fig. 5.


This work was supported by Foundation for National Key Basic Research Program of China (No. 2013CB910801 to L.M.), National Science Foundation of Hubei Province (No. 2013CFA133 to L.M.), High and New Technology Industrial Innovative Research Groups of the Wuhan Science and technology Bureau’s department (No. 2014CFA126, to L.M.).

Compliance with Ethical Standards

Conflict of interest

The authors declared that they have no conflicts of interest with the contents of this article.

Supplementary material

12033_2019_204_MOESM1_ESM.docx (840 kb)
Supplementary Material 1 (DOCX 840 kb)


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, College of Life SciencesHubei UniversityWuhanChina

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