Genetically Modified M13 Bacteriophage Nanonets for Enzyme Catalysis and Recovery

Enzyme-based biocatalysis exhibits multiple advantages over inorganic catalysts, including the biocompatibility and the unchallenged specificity of enzymes towards their substrate. The recovery and repeated use of enzymes is essential for any realistic application in biotechnology, but is not easily achieved with current strategies. For this purpose, enzymes are often immobilized on inorganic scaffolds, which could entail a reduction of the enzymes’ activity. Here, we show that immobilization to a nano-scaled biological scaffold, a nanonetwork of end-to-end cross-linked M13 bacteriophages, ensures high enzymatic activity and at the same time allows for the simple recovery of the enzymes. The bacteriophages have been genetically engineered to express AviTags at their ends, which permit biotinylation and their specific end-to-end self-assembly while allowing space on the major coat protein for enzyme coupling. We demonstrate that the phages form nanonetwork structures and that these so-called nanonets remain highly active even after re-using the nanonets multiple times in a flow-through reactor.

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M13 Bacteriophage-Based Self-Assembly Structures and Their Functional Capabilities

Mini-Reviews in Organic Chemistry ◽

10.2174/1570193x1203150429105418 ◽

2015 ◽

Vol 12 (3) ◽

pp. 271-281 ◽

Cited By ~ 21

Author(s):

Jong-Sik Moon ◽

Won-Geun Kim ◽

Chuntae Kim ◽

Geun-Tae Park ◽

Jeong Heo ◽

...

Keyword(s):

Self Assembly ◽

M13 Bacteriophage ◽

Functional Capabilities

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Formation of toroids by self-assembly of an α–α corner mimetic: supramolecular cyclization

Journal of Materials Chemistry B ◽

10.1039/c7tb01711a ◽

2017 ◽

Vol 5 (36) ◽

pp. 7583-7590 ◽

Cited By ~ 4

Author(s):

Debasish Podder ◽

Santu Bera ◽

Mintu Debnath ◽

Tanmay Das ◽

Debasish Haldar

Keyword(s):

Self Assembly ◽

Supramolecular Structure ◽

Cyclization Reaction ◽

End To End

An α–α corner mimetic self-assembles into a rod shape supramolecular structure which bends and closes end-to-end like a cyclization reaction to form toroids.

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Nanopore Formation by Self-Assembly of the Model Genetically Engineered Elastin-like Polymer [(VPGVG)2(VPGEG)(VPGVG)2]15

Journal of the American Chemical Society ◽

10.1021/ja047417f ◽

2004 ◽

Vol 126 (41) ◽

pp. 13212-13213 ◽

Cited By ~ 71

Author(s):

Javier Reguera ◽

Amir Fahmi ◽

Philip Moriarty ◽

Alessandra Girotti ◽

José Carlos Rodríguez-Cabello

Keyword(s):

Self Assembly ◽

Genetically Engineered

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Programmable Microbial Ink for 3D Printing of Living Materials Produced from Genetically Engineered Protein Nanofibers

10.1101/2021.04.19.440538 ◽

2021 ◽

Author(s):

Anna M Duraj-Thatte ◽

Avinash Manjula Basavanna ◽

Jarod Rutledge ◽

Jing Xia ◽

Shabir Hassan ◽

...

Keyword(s):

3D Printing ◽

Self Assembly ◽

Genetically Engineered ◽

Ambient Conditions ◽

Chemical Induction ◽

Genetic Circuits ◽

3D Print ◽

E Coli ◽

Molecular Components ◽

Living Materials

Living cells have the capability to synthesize molecular components and precisely assemble them from the nanoscale to build macroscopic living functional architectures under ambient conditions. The emerging field of living materials has leveraged microbial engineering to produce materials for various applications, but building 3D structures in arbitrary patterns and shapes has been a major challenge. We set out to develop a new bioink, termed as "microbial ink" that is produced entirely from genetically engineered microbial cells, programmed to perform a bottom-up, hierarchical self-assembly of protein monomers into nanofibers, and further into nanofiber networks that comprise extrudable hydrogels. We further demonstrate the 3D printing of functional living materials by embedding programmed Escherichia coli (E. coli) cells and nanofibers into microbial ink, which can sequester toxic moieties, release biologics and regulate its own cell growth through the chemical induction of rationally designed genetic circuits. This report showcases the advanced capabilities of nanobiotechnology and living materials technology to 3D-print functional living architectures.

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A nanostructured bifunctional acid–base catalyst resin formed by lyotropic liquid crystal monomers

Canadian Journal of Chemistry ◽

10.1139/cjc-2019-0408 ◽

2020 ◽

Vol 98 (7) ◽

pp. 332-336

Author(s):

Gregory E. Dwulet ◽

Benjamin J. Coscia ◽

Michael R. Shirts ◽

Douglas L. Gin

Keyword(s):

Liquid Crystal ◽

Self Assembly ◽

Polymeric Materials ◽

Dimethyl Acetal ◽

Bifunctional Catalyst ◽

Lyotropic Liquid Crystal ◽

Solid Catalyst ◽

Test Reaction ◽

Highly Active ◽

Nitroaldol Reaction

An ordered, nanoporous polymer resin was prepared from the self-assembly of lyotropic liquid crystal monomers and employed as a heterogeneous, bifunctional catalyst. This material contains antagonistic acid and base sites in the periodic nanopores and efficiently catalyzes a model tandem reaction (i.e., the deacetalization–nitroaldol reaction between benzaldehyde dimethyl acetal and nitromethane to yield β-nitrostyrene) with excellent product selectivity. This lyotropic liquid crystal-based solid catalyst represents one of the few examples of polymeric tandem catalysts synthesized by a “bottom-up” strategy that imparts control over the stoichiometry of acidic and basic monomers and is the only reported example of a lyotropic liquid crystal-based polymer that contains mutually incompatible catalytic groups. Furthermore, this heterogeneous catalyst is highly active, exhibiting a turnover frequency for this tandem test reaction that exceeds other reported catalytic polymeric materials.

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Experimental and numerical evaluation of a genetically engineered M13 bacteriophage with high sensitivity and selectivity for 2,4,6-trinitrotoluene

Organic & Biomolecular Chemistry ◽

10.1039/c8ob03075h ◽

2019 ◽

Vol 17 (23) ◽

pp. 5666-5670 ◽

Cited By ~ 2

Author(s):

Won-Geun Kim ◽

Chris Zueger ◽

Chuntae Kim ◽

Winnie Wong ◽

Vasanthan Devaraj ◽

...

Keyword(s):

Quantum Mechanics ◽

Numerical Evaluation ◽

High Sensitivity ◽

Genetically Engineered ◽

Experimental Results ◽

Sensor System ◽

M13 Bacteriophage ◽

Sensitivity And Selectivity

This study includes the experimental results of a sensitive M13 bacteriophage-based sensor system that are well matched with the quantum mechanics calculation.

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End-to-End Self-Assembly of RADA 16-I Nanofibrils in Aqueous Solutions

Biophysical Journal ◽

10.1016/j.bpj.2012.03.012 ◽

2012 ◽

Vol 102 (7) ◽

pp. 1617-1626 ◽

Cited By ~ 32

Author(s):

Paolo Arosio ◽

Marta Owczarz ◽

Hua Wu ◽

Alessandro Butté ◽

Massimo Morbidelli

Keyword(s):

Aqueous Solutions ◽

Self Assembly ◽

End To End

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Changes of biofilm properties in response to sorbed substances - an FTIR-ATR study

Water Science & Technology ◽

10.2166/wst.1995.0284 ◽

1995 ◽

Vol 32 (8) ◽

pp. 149-155 ◽

Cited By ~ 31

Author(s):

Jürgen Schmitt ◽

David Nivens ◽

David C. White ◽

Hans-Curt Flemming

Keyword(s):

Organic Pollutant ◽

Great Sensitivity ◽

Genetically Engineered ◽

Sorption Properties ◽

Environmental Properties ◽

Sterile Medium ◽

On Line ◽

Degradation Activity ◽

Flow Through ◽

Atr Spectroscopy

Biofilms play an important role as sorbents in the process of distribution of pollutants in surface water systems. The sorption properties of biofilms will influence the kind and the amount of sorbed substances. The heterogeneity of biofilms provides different sorption sites which exhibit a different sorption preference and capacity. As dynamic systems, biofilms will respond physiologically to their environment. Thus, the sorption of one substance may lead to a change in the composition of the EPS or other biofilm components and further alter the original sorption properties. In this paper, the influence of toluene on a biofilm was investigated. As a suitable method, FTIR-ATR spectroscopy was applied. The method is non destructive and allows the observation of biofilm formation and behaviour on line and in situ. A biofilm was allowed to form in ATR flow-through cells. The test strain was genetically engineered and contained a bioluminescent reporter gene which was switched on when toluene was metabolized. Thus, the degree of toluene degradation activity could be observed with great sensitivity. The FTIR spectrometer contained three flow-through cells which could be operated in parallel: one was run with sterile medium only, one with medium and bacteria, and one with medium, bacteria and toluene. This arrangement allowed the discrimination of the biofilm response from other effects. The IR spectrum showed specific bands of proteins, polysaccharides, phosphoryl compounds and other groups of molecules. A significant increase of EPS-polysaccharide formation was observed at a toluene level of 5 mg L−1. At 15 mg L−1, significantly more carboxyl groups were formed. Thus, the effect of the lipophilic organic pollutant toluene increased the amount of negatively charged groups and, consequently, the sorption capacity for metal cations. This result indicates that biofilms respond in a complex manner to different sorbates and alter their environmental properties.

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