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

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 cha...

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Bibliographic Details
Published in:Molecular biotechnology 2019-11, Vol.61 (11), p.816-825
Main Authors: Bian, Lu, Zhang, Zhen, Tang, Rong-xing, Shen, Wei, Ma, Li-xin
Format: Article
Language:English
Subjects:
Anchoring
Bacteria
Biocatalysis
Biochemistry
Biodegradation
Biodegradation, Environmental
Biological Techniques
Biotechnology
Catalysis
Catalytic activity
Cell Biology
Cell Membrane - metabolism
Cell surface
Cell Surface Display Techniques
Chemistry
Chemistry and Materials Science
E coli
Escherichia coli
Escherichia coli - enzymology
Escherichia coli - genetics
Escherichia coli - metabolism
Flavin
Flavin Mononucleotide
Fluorescence
Fusion protein
Human Genetics
Hydrolase
Hydrolases - genetics
Hydrolases - metabolism
Insecticides
Luminescent Proteins - chemistry
Luminescent Proteins - genetics
Methyl parathion
Methyl Parathion - metabolism
Organophosphorus compounds
Organophosphorus Compounds - metabolism
Original Paper
Parathion
Protein Science
Proteins
Recombinant Fusion Proteins
Substrates
Translocation
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Summary: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.
ISSN:1073-6085
1559-0305
DOI:10.1007/s12033-019-00204-3