Utilizing a divalent metal ion transporter to control biogenic nanoparticle synthesis.

Biogenic synthesis of inorganic nanomaterials has been demonstrated for both wild and engineered bacterial strains. In many systems the nucleation and growth of nanomaterials is poorly controlled and requires concentrations of heavy metals toxic to living cells. Here, we utilized the tools of synthetic biology to engineer a strain of Escherichia coli capable of synthesizing cadmium sulfide nanoparticles from low concentrations of reactants with control over the location of synthesis. Informed by simulations of bacterially-assisted nanoparticle synthesis, we created a strain of E. coli expressing a broad-spectrum divalent metal transporter, ZupT, and a synthetic CdS nucleating peptide. Expression of ZupT in the outer membrane and placement of the nucleating peptide in the periplasm focused synthesis within the periplasmic space and enabled sufficient nucleation and growth of nanoparticles at sub-toxic levels of the reactants. This strain synthesized internal CdS quantum dot nanoparticles with spherical morphology and an average diameter of approximately 3.3 nm. ONE-SENTENCE SUMMARY: Expression of a metal ion transporter regulates synthesis of cadmium sulfide nanoparticles in bacteria.

Light-Induced Patterning of Electroactive Bacterial Biofilms.

Electroactive bacterial biofilms can function as living biomaterials that merge the functionality of living cells with electronic components. However, the development of such advanced living electronics has been challenged by the inability to control the geometry of electroactive biofilms relative to solid-state electrodes. Here, we developed a lithographic strategy to pattern conductive biofilms of Shewanella oneidensis by controlling aggregation protein CdrAB expression with a blue light-induced genetic circuit. This controlled deposition enabled S. oneidensis biofilm patterning on transparent electrode surfaces, and electrochemical measurements allowed us to both demonstrate tunable conduction dependent on pattern size and quantify the intrinsic conductivity of the living biofilms. The intrinsic biofilm conductivity measurements enabled us to experimentally confirm predictions based on simulations of a recently proposed collision-exchange electron transport mechanism. Overall, we developed a facile technique for controlling electroactive biofilm formation on electrodes, with implications for both studying and harnessing bioelectronics.

Simulations to Aid in the Design of Microbes for Synthesis of Metallic Nanomaterials.

Microbes are champions of nanomaterial synthesis. By virtue of their incredible native range─from thermal vents to radioactive soil─microbes evolved tools to thrive on inorganic material, and, in their normal course of living, forge nanomaterials. In recent decades, synthetic biologists have engineered a vast array of functional nanomaterials using genetic tools that control the natural ability of bacteria to perform complex redox chemistry, maintain steep chemical gradients, and express biomolecular scaffolds. Leveraging microbial biology can lead to intricate nanomaterial architectures whose design and assembly exists beyond the ken of inorganic methods. Theories enumerating microbial nanomaterial synthesis are spare, however, despite the advantage they could offer. Here, we describe a theoretical approach to simulating biogenic nanomaterial synthesis that incorporates key features and parameters of Gram-negative bacteria. By adapting previously verified inorganic theories of nanoparticle synthesis, we recapitulate past biogenic experiments, such as the ability to localize nanoparticle synthesis or regulate nucleation of specific nanomaterials. Moreover, the simulation offers direction in the design of future experiments. Our results demonstrate the promise of marrying experimental and theoretical approaches to microbial nanomaterial synthesis.

Structure, self-assembly, and properties of a truncated reflectin variant.

Naturally occurring and recombinant protein-based materials are frequently employed for the study of fundamental biological processes and are often leveraged for applications in areas as diverse as electronics, optics, bioengineering, medicine, and even fashion. Within this context, unique structural proteins known as reflectins have recently attracted substantial attention due to their key roles in the fascinating color-changing capabilities of cephalopods and their technological potential as biophotonic and bioelectronic materials. However, progress toward understanding reflectins has been hindered by their atypical aromatic and charged residue-enriched sequences, extreme sensitivities to subtle changes in environmental conditions, and well-known propensities for aggregation. Herein, we elucidate the structure of a reflectin variant at the molecular level, demonstrate a straightforward mechanical agitation-based methodology for controlling this variant's hierarchical assembly, and establish a direct correlation between the protein's structural characteristics and intrinsic optical properties. Altogether, our findings address multiple challenges associated with the development of reflectins as materials, furnish molecular-level insight into the mechanistic underpinnings of cephalopod skin cells' color-changing functionalities, and may inform new research directions across biochemistry, cellular biology, bioengineering, and optics.

Computation in bacterial communities.

Bacteria across many scales are involved in a dynamic process of information exchange to coordinate activity and community structure within large and diverse populations. The molecular components bacteria use to communicate have been discovered and characterized, and recent efforts have begun to understand the potential for bacterial signal exchange to gather information from the environment and coordinate collective behaviors. Such computations made by bacteria to coordinate the action of a population of cells in response to information gathered by a multitude of inputs is a form of collective intelligence. These computations must be robust to fluctuations in both biological, chemical, and physical parameters as well as to operate with energetic efficiency. Given these constraints, what are the limits of computation by bacterial populations and what strategies have evolved to ensure bacterial communities efficiently work together? Here the current understanding of information exchange and collective decision making that occur in microbial populations will be reviewed. Looking toward the future, we consider how a deeper understanding of bacterial computation will inform future direction in microbiology, biotechnology, and biophysics.

Theoretical investigation of a genetic switch for metabolic adaptation.

Membrane transporters carry key metabolites across the cell membrane and, from a resource standpoint, are hypothesized to be produced when necessary. The expression of membrane transporters in metabolic pathways is often upregulated by the transporter substrate. In E. coli, such systems include for example the lacY, araFGH, and xylFGH genes, which encode for lactose, arabinose, and xylose transporters, respectively. As a case study of a minimal system, we build a generalizable physical model of the xapABR genetic circuit, which features a regulatory feedback loop via membrane transport (positive feedback) and enzymatic degradation (negative feedback) of an inducer. Dynamical systems analysis and stochastic simulations show that the membrane transport makes the model system bistable in certain parameter regimes. Thus, it serves as a genetic "on-off" switch, enabling the cell to only produce a set of metabolic enzymes when the corresponding metabolite is present in large amounts. We find that the negative feedback from the degradation enzyme does not significantly disturb the positive feedback from the membrane transporter. We investigate hysteresis in the switching and discuss the role of cooperativity and multiple binding sites in the model circuit. Fundamentally, this work explores how a stable genetic switch for a set of enzymes is obtained from transcriptional auto-activation of a membrane transporter through its substrate.

A dynamic thermoregulatory material inspired by squid skin.

Effective thermal management is critical for the operation of many modern technologies, such as electronic circuits, smart clothing, and building environment control systems. By leveraging the static infrared-reflecting design of the space blanket and drawing inspiration from the dynamic color-changing ability of squid skin, we have developed a composite material with tunable thermoregulatory properties. Our material demonstrates an on/off switching ratio of ~25 for the transmittance, regulates a heat flux of ~36 W/m2 with an estimated mechanical power input of ~3 W/m2, and features a dynamic environmental setpoint temperature window of ~8 °C. Moreover, the composite can manage one fourth of the metabolic heat flux expected for a sedentary individual and can also modulate localized changes in a wearer's body temperature by nearly 10-fold. Due to such functionality and associated figures of merit, our material may substantially reduce building energy consumption upon widespread deployment and adoption.

Self-Assembly of the Cephalopod Protein Reflectin.

Films from the cephalopod protein reflectin demonstrate multifaceted functionality as infrared camouflage coatings, proton transport media, and substrates for growth of neural stem cells. A detailed study of the in vitro formation, structural characteristics, and stimulus response of such films is presented. The reported observations hold implications for the design and development of advanced cephalopod-inspired functional materials.

Oxidation of Ethidium using TAML Activators: A Model for High School Research Performed in Partnership with University Scientists.

A chemical research program at a public high school has been developed. The full-year Advanced Chemical Research class (ACR) in the high school enrolls 20 to 30 seniors each year, engaging them in long-term experimental projects. Through partnerships involving university scientists, ACR high school students have had the opportunity to explore a number of highly sophisticated original research projects. As an example of the quality of experimental work made possible through these high school-university partnerships, this article describes the development of a novel method for the oxidation of ethidium bromide, a mutagen commonly used in molecular biology. Data collected from ACR alumni show that the ACR program is instrumental in encouraging students to pursue careers in scientific fields and in creating life-long problem-solvers.