Microbially induced calcium carbonate precipitation through CO2 sequestration via an engineered Bacillus subtilis.

BACKGROUND: Microbially induced calcium carbonate precipitation has been extensively researched for geoengineering applications as well as diverse uses within the built environment. Bacteria play a crucial role in producing calcium carbonate minerals, via enzymes including carbonic anhydrase-an enzyme with the capability to hydrolyse CO2, commonly employed in carbon capture systems. This study describes previously uncharacterised carbonic anhydrase enzyme sequences capable of sequestering CO2 and subsequentially generating CaCO3 biominerals and suggests a route to produce carbon negative cementitious materials for the construction industry. RESULTS: Here, Bacillus subtilis was engineered to recombinantly express previously uncharacterised carbonic anhydrase enzymes from Bacillus megaterium and used as a whole cell catalyst allowing this novel bacterium to sequester CO2 and convert it to calcium carbonate. A significant decrease in CO2 was observed from 3800 PPM to 820 PPM upon induction of carbonic anhydrase and minerals recovered from these experiments were identified as calcite and vaterite using X-ray diffraction. Further experiments mixed the use of this enzyme (as a cell free extract) with Sporosarcina pasteurii to increase mineral production whilst maintaining a comparable level of CO2 sequestration. CONCLUSION: Recombinantly produced carbonic anhydrase successfully sequestered CO2 and converted it into calcium carbonate minerals using an engineered microbial system. Through this approach, a process to manufacture cementitious materials with carbon sequestration ability could be developed.

Effect of (in)organic additives on microbially induced calcium carbonate precipitation.

AIM: This study aimed to understand the morphological effects of (in)organic additives on microbially induced calcium carbonate precipitation (MICP). METHODS AND RESULTS: MICP was monitored in real time in the presence of (in)organic additives: bovine serum albumin (BSA), biofilm surface layer protein A (BslA), magnesium chloride (MgCl2), and poly-l-lysine. This monitoring was carried out using confocal microscopy to observe the formation of CaCO3 from the point of nucleation, in comparison to conditions without additives. Complementary methodologies, namely scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction, were employed to assess the visual morphology, elemental composition, and crystalline structures of CaCO3, respectively, following the crystals' formation. The results demonstrated that in the presence of additives, more CaCO3 crystals were produced at 100 min compared to the reaction without additives. The inclusion of BslA resulted in larger crystals than reactions containing other additives, including MgCl2. BSA induced a significant number of crystals from the early stages of the reaction (20 min) but did not have a substantial impact on crystal size compared to conditions without additives. All additives led to a higher content of calcite compared to vaterite after a 24-h reaction, with the exception of MgCl2, which produced a substantial quantity of magnesium calcite. CONCLUSIONS: The work demonstrates the effect of several (in)organic additives on MICP and sets the stage for further research to understand additive effects on MICP to achieve controlled CaCO3 precipitation.

Biological, physical and morphological factors for the programming of a novel microbial hygromorphic material.

The urgency for energy efficient, responsive architectures has propelled smart material development to the forefront of scientific and architectural research. This paper explores biological, physical, and morphological factors influencing the programming of a novel microbial-based smart hybrid material which is responsive to changes in environmental humidity. Hygromorphs respond passively, without energy input, by expanding in high humidity and contracting in low humidity.Bacillus subtilisdevelops environmentally robust, hygromorphic spores which may be harnessed within a bilayer to generate a deflection response with potential for programmability. The bacterial spore-based hygromorph biocomposites (HBCs) were developed and aggregated to enable them to open and close apertures and demonstrate programmable responses to changes in environmental humidity. This study spans many fields including microbiology, materials science, design, fabrication and architectural technology, working at multiple scales from single cells to 'bench-top' prototype.Exploration of biological factors at cellular and ultracellular levels enabled optimisation of growth and sporulation conditions to biologically preprogramme optimum spore hygromorphic response and yield. Material explorations revealed physical factors influencing biomechanics, preprogramming shape and response complexity through fabrication and inert substrate interactions, to produce a palette of HBCs. Morphological aggregation was designed to harness and scale-up the HBC palette into programmable humidity responsive aperture openings. This culminated in pilot performance testing of a humidity-responsive ventilation panel fabricated with aggregatedBacillusHBCs as a bench-top prototype and suggests potential for this novel biotechnology to be further developed.

Biofilm inspired fabrication of functional bacterial cellulose through ex-situ and in-situ approaches.

Bacterial cellulose (BC) has been explored for use in a range of applications including tissue engineering and textiles. BC can be produced from waste streams, but sustainable approaches are needed for functionalisation. To this end, BslA, a B. subtilis biofilm protein was produced recombinantly with and without a cellulose binding module (CBM) and the cell free extract was used to treat BC either ex-situ, through drip coating or in-situ, by incorporating during fermentation. The results showed that ex-situ modified BC increased the hydrophobicity and water contact angle reached 120°. In-situ experiments led to a BC film morphological change and mechanical testing demonstrated that addition of BslA with CBM resulted in a stronger, more elastic material. This study presents a nature inspired approach to functionalise BC using a biofilm hydrophobin, and we demonstrate that recombinant proteins could be effective and sustainable molecules for functionalisation of BC materials.

Innovating fire safety with recombinant hydrophobic proteins for textile fire retardancy.

Fire retardancy for textiles is important to prevent the rapid spread of fire and minimize damage to property and harm to human life. To infer fire-resistance on textile materials such as cotton or nylon, chemical coatings are often used. These chemicals are usually toxic, and economically and environmentally unsustainable, however, some naturally produced protein-based fire retardants could be an alternative. A biofilm protein from Bacillus subtilis (BslA) was identified and recombinantly expressed in Escherichia coli with a double cellulose binding domain. It was then applied to a range of natural and synthetic fabric materials. A flame retardancy test found that use of BslA reduced fire damage by up to 51% and would pass fire retardancy testing according to British standards. It is therefore a viable and sustainable alternative to current industrial fire-retardant coatings.

Are Mushrooms Parametric?

Designing with biological materials as a burgeoning approach in the architecture field requires the development of new design strategies and fabrication methods. In this paper, we question if designers can use a parametric design approach while working with living materials. The research uses fungi as a biomaterial probe to experiment with the parametric behavior of living systems. Running design experiments using fungi helps to understand the extent to which biological systems can be considered parametric and, if so, what kind of parametric systems they are. Answering these questions provides a method to work with complex biological systems and may lead to new approaches of fabricating materials by tuning the environmental parameters of biological growth.

Mild hydrostatic pressure triggers oxidative responses in Escherichia coli.

Hydrostatic pressure is an important physical stimulus which can cause various responses in bacterial cells. The survival and cellular processes of Escherichia coli under hydrostatic pressures between 10 MPa and 110 MPa have been studied. However, understanding bacterial responses to moderately elevated pressure of up to 10 MPa is useful for a range of different applications including for example in smart and responsive materials. In this study, the genetic responses of E. coli K-12 MG1655 to 1 MPa pressure was examined using transcriptomic analysis by RNA-Seq. The results show that 101 genes were differentially expressed under 1 MPa pressure in E. coli cells, with 85 of them up-regulated. The analysis suggested that some genes were over expressed to adapt the increase of oxygen levels in our system, and several functional categories are involved including oxidative stress responses, Fe-S cluster assembly and iron acquisition. Two differentially expressed genes azuC and entC were further investigated using RT-qPCR, and GFP reported strains of those two genes were created, AG1319 (PazuC azuC-msfgfp) and AG1321 (PentC entC-msfgfp). A linear response of azuC expression was observed between 0 MPa to 1 MPa by monitoring the fluorescence signal of strain AG1319 (PazuC azuC-msfgfp). This study is the first report to demonstrate the genetic response of bacterial cells under 1 MPa hydrostatic pressure, and provides preliminary data for creating pressure sensing bacterial strains for a wide range of applications.

Design and modelling of an engineered bacteria-based, pressure-sensitive soil.

In this paper, we describe the first steps in the design of a synthetic biological system based on the use of genetically modified bacteria to detect elevated pressures in soils and respond by cementing soil particles. Such a system might, for example, enable a self- constructed foundation to form in response to load using engineered bacteria which could be seeded and grown in the soils. This process would reduce the need for large-scale excavations and may be the basis for a new generation of self-assembling and responsive bio-based materials. A prototype computational model is presented which integrates experimental data from a pressure sensitive gene within Escherichia coli bacteria with geotechnical models of soil loading and pore water pressure. The results from the integrated model are visualised by mapping expected gene expression values onto the soil volume. We also use our experimental data to design a two component system where one type of bacteria acts as a sensor and signals to another material synthesis bacteria. The simulation demonstrates the potential of computational models which integrate multiple scales from macro stresses in soils to the expression of individual genes to inform new types of design process. The work also illustrates the combination of in silico (silicon based computing) computation with in vivo (in the living) computation.

Architects of nature: growing buildings with bacterial biofilms.

In his text 'On Architecture', Vitruvius suggested that architecture is an imitation of nature. Here we discuss what happens when we begin using nature in architecture. We describe recent developments in the study of biofilm structure, and propose combining modern architecture and synthetic microbiology to develop sustainable construction approaches. Recently, Kolodkin-Gal laboratory and others revealed a role for precipitation of calcium carbonate in the maturation and assembly of bacterial communities with complex structures. Importantly, they demonstrated that different secreted organic materials shape the calcium carbonate crystals formed by the bacterial cells. This provides a proof-of-concept for a potential use of bacteria in designing rigid construction materials and altering crystal morphology and function. In this study, we discuss how these recent discoveries may change the current strategies of architecture and construction. We believe that biofilm communities enhanced by synthetic circuits may be used to construct buildings and to sequester carbon dioxide in the process.

AMUC: Associated Motion capture User Categories.

The AMUC (Associated Motion capture User Categories) project consisted of building a prototype sketch retrieval client for exploring motion capture archives. High-dimensional datasets reflect the dynamic process of motion capture and comprise high-rate sampled data of a performer's joint angles; in response to multiple query criteria, these data can potentially yield different kinds of information. The AMUC prototype harnesses graphic input via an electronic tablet as a query mechanism, time and position signals obtained from the sketch being mapped to the properties of data streams stored in the motion capture repository. As well as proposing a pragmatic solution for exploring motion capture datasets, the project demonstrates the conceptual value of iterative prototyping in innovative interdisciplinary design. The AMUC team was composed of live performance practitioners and theorists conversant with a variety of movement techniques, bioengineers who recorded and processed motion data for integration into the retrieval tool, and computer scientists who designed and implemented the retrieval system and server architecture, scoped for Grid-based applications. Creative input on information system design and navigation, and digital image processing, underpinned implementation of the prototype, which has undergone preliminary trials with diverse users, allowing identification of rich potential development areas.