Expression of xylanase on Escherichia coli using a truncated ice nucleation protein of Erwinia ananas (InaA)

2019 ◽  
Vol 78 ◽  
pp. 25-32 ◽  
Author(s):  
Mei Yuin Joanne Wee ◽  
Abdul Munir Abd. Murad ◽  
Farah Diba Abu Bakar ◽  
Kheng Oon Low ◽  
Rosli Md Illias
Keyword(s):  
Escherichia Coli ◽  
Ice Nucleation ◽  
Ice Nucleation Protein ◽  
Erwinia Ananas

Freezing from the inside: Ice nucleation in Escherichia coli and Escherichia coli ghosts by inner membrane bound ice nucleation protein InaZ

2020 ◽  
Vol 15 (3) ◽  
pp. 031003
Author(s):  
Johannes Kassmannhuber ◽  
Sergio Mauri ◽  
Mascha Rauscher ◽  
Nadja Brait ◽  
Lea Schöner ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ice Nucleation ◽  
Ice Nucleation Protein ◽  
Inner Membrane ◽  
Membrane Bound

Expression of Carboxymethylcellulase on the Surface of Escherichia Coli Using Pseudomonas Syringae Ice Nucleation Protein

1998 ◽  
Vol 22 (5) ◽  
pp. 348-354 ◽  
Author(s):  
Heung-Chae Jung ◽  
Joon-Hyun Park ◽  
Seung-Hwan Park ◽  
Jean-Michel Lebeault ◽  
Jae-Gu Pan
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Syringae ◽  
Ice Nucleation ◽  
Ice Nucleation Protein

scholarly journals Cell Surface Display of Human Immunodeficiency Virus Type 1 gp120 on Escherichia coli by Using Ice Nucleation Protein

1999 ◽  
Vol 6 (4) ◽  
pp. 499-503 ◽  
Author(s):  
Young-Don Kwak ◽  
Seung-Ku Yoo ◽  
Eui-Joong Kim
Keyword(s):  
Escherichia Coli ◽  
Human Immunodeficiency Virus ◽  
Surface Display ◽  
Cell Surface Display ◽  
Viral Proteins ◽  
Ice Nucleation ◽  
Ice Nucleation Protein ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT A new system designed for cell surface display of recombinant proteins on Escherichia coli has been evaluated for expression of eukaryotic viral proteins. Human immunodeficiency virus type 1 (HIV-1) gp120 was fused to the C terminus of ice nucleation protein (INP), an outer membrane protein of Pseudomonas syringae. Western blotting, immunofluorescence microscopy, fluorescence-activated cell-sorting analysis, whole-cell enzyme-linked immunosorbent assay, and ice nucleation activity assay confirmed the successful expression of HIV-1 gp120 on the surface ofEscherichia coli. This study shows that the INP system can be used for the expression of eukaryotic viral proteins. There is also a possibility that the INP system can be used as an AIDS diagnostic system, an oral vaccine delivery system, and an expression system for various heterologous higher-molecular-weight proteins.

scholarly journals Cotranslocation of Methyl Parathion Hydrolase to the Periplasm and of Organophosphorus Hydrolase to the Cell Surface of Escherichia coli by the Tat Pathway and Ice Nucleation Protein Display System

2009 ◽  
Vol 76 (2) ◽  
pp. 434-440 ◽  
Author(s):  
Chao Yang ◽  
Roland Freudl ◽  
Chuanling Qiao ◽  
Ashok Mulchandani
Keyword(s):  
Escherichia Coli ◽  
Methyl Parathion ◽  
Ice Nucleation ◽  
Organophosphorus Hydrolase ◽  
Display System ◽  
Ice Nucleation Protein ◽  
Methyl Parathion Hydrolase ◽  
E Coli ◽  
Tat Pathway ◽  
Parathion Hydrolase

ABSTRACT A genetically engineered Escherichia coli strain coexpressing organophosphorus hydrolase (OPH) and methyl parathion hydrolase (MPH) was constructed for the first time by cotransforming two compatible plasmids. Since these two enzymes have different substrate specificities, the coexpression strain showed a broader substrate range than strains expressing either one of the hydrolases. To reduce the mass transport limitation of organophosphates (OPs) across the cell membrane, MPH and OPH were simultaneously translocated to the periplasm and cell surface of E. coli, respectively, by employing the twin-arginine translocation (Tat) pathway and ice nucleation protein (INP) display system. The resulting recombinant strain showed sixfold-higher whole-cell activity than the control strain expressing cytosolic OP hydrolases. The correct localization of MPH and OPH was demonstrated by cell fractionation, immunoblotting, and enzyme activity assays. No growth inhibition was observed for the recombinant E. coli strain, and suspended cultures retained almost 100% of the activity over a period of 2 weeks. Owing to its high level of activity and superior stability, the recombinant E. coli strain could be employed as a whole-cell biocatalyst for detoxification of OPs. This strategy of utilizing dual translocation pathways should open up new avenues for cotranslocating multiple functional moieties to different extracytosolic compartments of a bacterial cell.

scholarly journals Whole-cell catalysis by surface display of fluorinase on Escherichia coli using N-terminal domain of ice nucleation protein

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Xinming Feng ◽  
Miaomiao Jin ◽  
Wei Huang ◽  
Wei Liu ◽  
Mo Xian
Keyword(s):  
Escherichia Coli ◽  
Catalytic Activity ◽  
Industrial Application ◽  
Large Scale ◽  
Surface Display ◽  
Ice Nucleation ◽  
Ice Nucleation Protein ◽  
Whole Cell ◽  
Whole Cell Catalysis

Abstract Background Fluorinases play a unique role in the production of fluorine-containing organic molecules by biological methods. Whole-cell catalysis is a better choice in the large-scale fermentation processes, and over 60% of industrial biocatalysis uses this method. However, the in vivo catalytic efficiency of fluorinases is stuck with the mass transfer of the substrates. Results A gene sequence encoding a protein with fluorinase function was fused to the N-terminal of ice nucleation protein, and the fused fluorinase was expressed in Escherichia coli BL21(DE3) cells. SDS-PAGE and immunofluorescence microscopy were used to demonstrate the surface localization of the fusion protein. The fluorinase displayed on the surface showed good stability while retaining the catalytic activity. The engineered E.coli with surface-displayed fluorinase could be cultured to obtain a larger cell density, which was beneficial for industrial application. And 55% yield of 5′-fluorodeoxyadenosine (5′-FDA) from S-adenosyl-L-methionine (SAM) was achieved by using the whole-cell catalyst. Conclusions Here, we created the fluorinase-containing surface display system on E.coli cells for the first time. The fluorinase was successfully displayed on the surface of E.coli and maintained its catalytic activity. The surface display provides a new solution for the industrial application of biological fluorination. Graphical Abstract

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