Towards area‐based in vitro metabolic engineering: Assembly of Pfs enzyme onto patterned microfabricated chips

Lewandowski, Angela; Bentley, William E.; Yi, Hyunmin; Rubloff, Gary W.; Payne, Gregory F.; Ghodssi, Reza

doi:10.1002/btpr.44

Cited by 18 publications

(18 citation statements)

References 37 publications

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“…Specifically, in Fig. 5b , A and B populations were incubated at mid-exponential phase with in vitro -synthesized AI-2 (refs 50 , 51 ) at concentrations: 0, 2, 10, 28 and 75 μM. After 12 h, samples were observed for fluorescence by confocal microscopy and then quantified by fluorescence-activated cell sorting (FACS; Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Nano-guided cell networks as conveyors of molecular communication

Terrell

Tsao

et al. 2015

Nat Commun

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Advances in nanotechnology have provided unprecedented physical means to sample molecular space. Living cells provide additional capability in that they identify molecules within complex environments and actuate function. We have merged cells with nanotechnology for an integrated molecular processing network. Here we show that an engineered cell consortium autonomously generates feedback to chemical cues. Moreover, abiotic components are readily assembled onto cells, enabling amplified and ‘binned' responses. Specifically, engineered cell populations are triggered by a quorum sensing (QS) signal molecule, autoinducer-2, to express surface-displayed fusions consisting of a fluorescent marker and an affinity peptide. The latter provides means for attaching magnetic nanoparticles to fluorescently activated subpopulations for coalescence into colour-indexed output. The resultant nano-guided cell network assesses QS activity and conveys molecular information as a ‘bio-litmus' in a manner read by simple optical means.

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Section: Resultsmentioning

confidence: 99%

Nano-guided cell networks as conveyors of molecular communication

Terrell

Tsao

et al. 2015

Nat Commun

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“…Figure 12a shows that tyrosinase converts the phenolic tyrosine residues of proteins to o-quinone moieties that can undergo subsequent uncatalyzed reactions to covalently conjugate the protein to chitosan (see below for details of quinone-chitosan reactions). Tyrosinase has been used to mediate the grafting of peptides [182], open-chain proteins (e.g., gelatin [183,184] and silk proteins [185][186][187][188][189][190]) and proteins with globular structures (e.g., fluorescent proteins [191], binding proteins [140] and enzymes [141,[192][193][194]) to chitosan. For the case of globular proteins, the tyrosine residues may be inaccessible (i.e., tyrosinase cannot access these residues and convert them into o-quinones).…”

Section: Biofunctionalizing Electrodeposited Chitosan Filmsmentioning

confidence: 99%

Chitosan to Connect Biology to Electronics: Fabricating the Bio-Device Interface and Communicating Across This Interface

et al. 2014

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Individually, advances in microelectronics and biology transformed the way we live our lives. However, there remain few examples in which biology and electronics have been interfaced to create synergistic capabilities. We believe there are two major challenges to the integration of biological components into microelectronic systems: (i) assembly of the biological components at an electrode address, and (ii) communication between the assembled biological components and the underlying electrode. Chitosan OPEN ACCESSPolymers 2015, 7 2 possesses a unique combination of properties to meet these challenges and serve as an effective bio-device interface material. For assembly, chitosan's pH-responsive film-forming properties allow it to "recognize" electrode-imposed signals and respond by self-assembling as a stable hydrogel film through a cathodic electrodeposition mechanism. A separate anodic electrodeposition mechanism was recently reported and this also allows chitosan hydrogel films to be assembled at an electrode address. Protein-based biofunctionality can be conferred to electrodeposited films through a variety of physical, chemical and biological methods. For communication, we are investigating redox-active catechol-modified chitosan films as an interface to bridge redox-based communication between biology and an electrode. Despite significant progress over the last decade, many questions still remain which warrants even deeper study of chitosan's structure, properties, and functions.

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“…To facilitate the enzymatic conjugation of globular proteins (e.g., enzymes), one can genetically engineer the protein to have a short amino acids sequence (i.e., a fusion tag at the C‐ or N‐terminals) that is exposed for the enzyme . For example, tyrosine residues have been genetically “fused” to enhance the tyrosinase‐mediated protein conjugation on chitosan . As will be discussed in Section , a model enzyme with tyrosine fusion tags was immobilized on a chitosan‐coated chip to allow for electrical signal triggered attenuation of enzyme activity …”

Section: Electroaddressable Thin Hydrogel Films To Facilitate Signal mentioning

confidence: 99%

Connecting Biology to Electronics: Molecular Communication via Redox Modality

Liu

Tschirhart

et al. 2017

Adv Healthcare Materials

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Biology and electronics are both expert at for accessing, analyzing, and responding to information. Biology uses ions, small molecules, and macromolecules to receive, analyze, store, and transmit information, whereas electronic devices receive input in the form of electromagnetic radiation, process the information using electrons, and then transmit output as electromagnetic waves. Generating the capabilities to connect biology-electronic modalities offers exciting opportunities to shape the future of biosensors, point-of-care medicine, and wearable/implantable devices. Redox reactions offer unique opportunities for bio-device communication that spans the molecular modalities of biology and electrical modality of devices. Here, an approach to search for redox information through an interactive electrochemical probing that is analogous to sonar is adopted. The capabilities of this approach to access global chemical information as well as information of specific redox-active chemical entities are illustrated using recent examples. An example of the use of synthetic biology to recognize external molecular information, process this information through intracellular signal transduction pathways, and generate output responses that can be detected by electrical modalities is also provided. Finally, exciting results in the use of redox reactions to actuate biology are provided to illustrate that synthetic biology offers the potential to guide biological response through electrical cues.

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Towards area‐based in vitro metabolic engineering: Assembly of Pfs enzyme onto patterned microfabricated chips

Cited by 18 publications

References 37 publications

Nano-guided cell networks as conveyors of molecular communication

Nano-guided cell networks as conveyors of molecular communication

Chitosan to Connect Biology to Electronics: Fabricating the Bio-Device Interface and Communicating Across This Interface

Connecting Biology to Electronics: Molecular Communication via Redox Modality

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