Complete genome sequence and transcriptome response to vitamin C supplementation of Novacetimonas hansenii SI1 - producer of highly-stretchable cellulose.

Novacetimonas hansenii SI1, previously known as Komagataeibacter hansenii, produces bacterial nanocellulose (BNC) with unique ability to stretch. The addition of vitamin C in the culture medium increases the porosity of the membranes and their stretchability making them highly moldable. To better understand the genetic background of this strain, we obtained its complete genome sequence using a hybrid sequencing and assembly strategy. We described the functional regions in the genome which are important for the synthesis of BNC and acetan-like II polymer. We next investigated the effect of 1% vitamin C supplementation on the global gene expression profile using RNA sequencing. Our transcriptomic readouts imply that vitamin C functions mainly as a reducing agent. We found that the changes in cellular redox status are balanced by strong repression of the sulfur assimilation pathway. Moreover, in the reduced conditions, glucose oxidation is decreased and alternative pathways for energy generation, such as acetate accumulation, are activated. The presence of vitamin C negatively influences acetan-like II polymer biosynthesis, which may explain the lowered yield and changed mechanical properties of BNC. The results of this study enrich the functional characteristics of the genomes of the efficient producers of the N. hansenii species. Improved understanding of the adaptation to the presence of vitamin C at the molecular level has important guiding significance for influencing the biosynthesis of BNC and its morphology.

Synergistic outcomes of Chlorella-bacterial cellulose based hydrogel as an ethylene scavenger.

Increasing the freshness of vegetables requires the elimination of ethylene, which can be done through chemical methods. However, the development of eco-friendly approaches is required for environmental reasons. Chlorella vulgaris (C. vulgaris) was selected as a new biological material for demonstrating an excellent performance in ethylene removal. To support C. vulgaris, bacterial cellulose (BC) produced by Gluconacetobacter hansenii (G. hansenii) was chosen due to its high water content and biodegradability. To increase BC productivity, UV-induced mutant G. hansenii was isolated, and they produced high yields of BC (9.80 ±â€¯0.52 g/L). Furthermore, comparative transcriptome analysis revealed metabolic flux changes toward UDP-glucose accumulation and enhanced BC production. BC-based hydrogels (BC hydrogels) were successfully prepared using a 2.4 % carboxymethyl cellulose (CMC) and 1 % agar mixture. We used Chlorella-BC hydrogels as an ethylene scavenger, which reduced 90 % of ethylene even when the immobilized C. vulgaris was preserved for 14 days at room temperature without media supplementation. We demonstrated for the first time the potential of BC hydrogels to integrate C. vulgaris as a sustainable ethylene absorber for green food packaging and biomass technology.

Hybrid Silver-Containing Materials Based on Various Forms of Bacterial Cellulose: Synthesis, Structure, and Biological Activity.

Sustained interest in the use of renewable resources for the production of medical materials has stimulated research on bacterial cellulose (BC) and nanocomposites based on it. New Ag-containing nanocomposites were obtained by modifying various forms of BC with Ag nanoparticles prepared by metal-vapor synthesis (MVS). Bacterial cellulose was obtained in the form of films (BCF) and spherical BC beads (SBCB) by the Gluconacetobacter hansenii GH-1/2008 strain under static and dynamic conditions. The Ag nanoparticles synthesized in 2-propanol were incorporated into the polymer matrix using metal-containing organosol. MVS is based on the interaction of extremely reactive atomic metals formed by evaporation in vacuum at a pressure of 10-2 Pa with organic substances during their co-condensation on the cooled walls of a reaction vessel. The composition, structure, and electronic state of the metal in the materials were characterized by transmission and scanning electron microscopy (TEM, SEM), powder X-ray diffraction (XRD), small-angle X-ray scattering (SAXS) and X-ray photoelectron spectroscopy (XPS). Since antimicrobial activity is largely determined by the surface composition, much attention was paid to studying its properties by XPS, a surface-sensitive method, at a sampling depth about 10 nm. C 1s and O 1s spectra were analyzed self-consistently. XPS C 1s spectra of the original and Ag-containing celluloses showed an increase in the intensity of the C-C/C-H groups in the latter, which are associated with carbon shell surrounding metal in Ag nanoparticles (Ag NPs). The size effect observed in Ag 3d spectra evidenced on a large proportion of silver nanoparticles with a size of less than 3 nm in the near-surface region. Ag NPs in the BC films and spherical beads were mainly in the zerovalent state. BC-based nanocomposites with Ag nanoparticles exhibited antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, Escherichia coli bacteria and Candida albicans and Aspergillus niger fungi. It was found that AgNPs/SBCB nanocomposites are more active than Ag NPs/BCF samples, especially against Candida albicans and Aspergillus niger fungi. These results increase the possibility of their medical application.

Structural properties of optically clear bacterial cellulose produced by Komagataeibacter hansenii using arabitol.

Bacterial cellulose (BC) exhibits beneficial properties for use in biomedical applications but is limited by its lack of tunable transparency capabilities. To overcome this deficiency, a novel method to synthesize transparent BC materials using an alternative carbon source, namely arabitol, was developed. Characterization of the BC pellicles was performed for yield, transparency, surface morphology, and molecular assembly. Transparent BC was produced using mixtures of glucose and arabitol. Zero percent arabitol pellicles exhibited 25% light transmittance, which increased with increasing arabitol concentration through to 75% light transmittance. While transparency increased, overall BC yield was maintained indicating that the altered transparency may be induced on a micro-scale rather than a macro-scale. Significant differences in fiber diameter and the presence of aromatic signatures were observed. Overall, this research outlines methods for producing BC with tunable optical transparency, while also bringing new insight to insoluble components of exopolymers produced by Komagataeibacter hansenii.

Analysis of cellulose synthesis in a high-producing acetic acid bacterium Komagataeibacter hansenii.

Bacterial cellulose (BC) represents a renewable biomaterial with unique properties promising for biotechnology and biomedicine. Komagataeibacter hansenii ATCC 53,582 is a well-characterized high-yield producer of BC used in the industry. Its genome encodes three distinct cellulose synthases (CS), bcsAB1, bcsAB2, and bcsAB3, which together with genes for accessory proteins are organized in operons of different complexity. The genetic foundation of its high cellulose-producing phenotype was investigated by constructing chromosomal in-frame deletions of the CSs and of two predicted regulatory diguanylate cyclases (DGC), dgcA and dgcB. Proteomic characterization suggested that BcsAB1 was the decisive CS because of its high expression and its exclusive contribution to the formation of microcrystalline cellulose. BcsAB2 showed a lower expression level but contributes significantly to the tensile strength of BC and alters fiber diameter significantly as judged by scanning electron microscopy. Nevertheless, no distinct extracellular polymeric substance (EPS) from this operon was identified after static cultivation. Although transcription of bcsAB3 was observed, expression of the protein was below the detection limit of proteome analysis. Alike BcsAB2, deletion of BcsAB3 resulted in a visible reduction of the cellulose fiber diameter. The high abundance of BcsD and the accessory proteins CmcAx, CcpAx, and BglxA emphasizes their importance for the proper formation of the cellulosic network. Characterization of deletion mutants lacking the DGC genes dgcA and dgcB suggests a new regulatory mechanism of cellulose synthesis and cell motility in K. hansenii ATCC 53,582. Our findings form the basis for rational tailoring of the characteristics of BC. KEY POINTS: • BcsAB1 induces formation of microcrystalline cellulose fibers. • Modifications by BcsAB2 and BcsAB3 alter diameter of cellulose fibers. • Complex regulatory network of DGCs on cellulose pellicle formation and motility.

Bacterial cellulose production by Komagataeibacter hansenii can be improved by successive batch culture.

Bacterial cellulose (BC) is a biopolymer principally synthetized by strains of the genus Komagataeibacter. However, high costs and low production yield make large-scale application difficult. The aim of this work was to evaluate the effects of successive batch culture before fermentation on the ability to increase the capacity of bacterial cellulose biosynthesis by a low-producing strain. The Komagataeibacter hansenii strain ATCC 23,769 was initially cultivated in fermentation broth for two periods of 35 or 56 days under static conditions. At the end of each period of time, they were transferred to new broth to be cultivated again (new batch culture cycle) for 35 or 56 days and carried out in parallel with a 10-day fermentation to determine the quantity of BC produced. As a result, a greater increase was observed after the end of the second and third batch cultures of 56 days (increases of 137% and 187% in relation to the nonbatch cultured strain, respectively). The produced samples presented higher crystallinity and thermal properties but lower water holding capacity. Through this work, it was concluded that the longer the batch culture time was, the greater the increase in the capacity of cellulose biosynthesis, which also depended on the number of successive batch culture cycles carried out.

Bacterial cellulose production by Novacetimonas hansenii MSCL 1646 on apple juice.

Biomaterials and biopolymers, such as bacterial cellulose (BC), are becoming increasingly important as sustainable materials with a wide range of potential applications. However, BC industrial production is associated with several difficulties such as low BC production yields and high production costs; therefore, cheap alternative growth media, e.g. apple juice are being studied intensively. The aim of this study is to evaluate BC synthesis under static conditions on apple juice medium (AJM). The optimal concentration of apple juice in unsupplemented AJM for Novacetimonas hansenii MSCL 1646 was shown by its dilution 1:6 with water, which resulted in 0.89 ± 0.01 g/L of dry BC weight after 10 cultivation days. Low BC synthesis can be associated with insufficient N concentration in apple juice; therefore, different organic and inorganic N sources were evaluated in combination with AJM, and beef extract (5 g/L) was found to be the most suitable. Further, AJM optimisation experiment showed the optimal apple juice and beef extract concentrations as 1:2 and 15 g/L respectively, which resulted in 17.27 ± 0.07 g/L of dry BC weight, which is significantly higher than in standard Hestrin-Schramm (HS) medium (4.07 ± 0.02 g/L). Analysis of mechanical and physical properties showed that use of AJM results in changes in BC properties compared with the standard HS medium. Results of the study indicate that apple juice is an effective and cheap C source that in combination with appropriate N source leads to high BC synthesis and makes it suitable for industrial BC production. KEY POINTS: • Low quality apples can be used as raw material for BC production; • Beef extract improves BC synthesis in apple juice medium; • Use of apple juice and beef extract affect mechanical properties of BC.

Metagenome-Assembled Genomes of Komagataeibacter from Kombucha Exposed to Mars-Like Conditions Reveal the Secrets in Tolerating Extraterrestrial Stresses.

Kombucha mutualistic community (KMC) is composed by acetic acid bacteria and yeasts, producing fermented tea with health benefits. As part of the BIOlogy and Mars EXperiment (BIOMEX) project, the effect of Mars-like conditions on the KMC was analyzed. Here, we analyzed metagenome-assembled genomes (MAGs) of the Komagataeibacter, which is a predominant genus in KMC, to understand their roles in the KMC after exposure to Mars-like conditions (outside the International Space Station) based on functional genetic elements. We constructed three MAGs: K. hansenii, K. rhaeticus, and K. oboediens. Our results showed that (i) K. oboediens MAG functionally more complex than K. hansenii, (ii) K. hansenii is a keystone in KMCs with specific functional features to tolerate extreme stress, and (iii) genes related to the PPDK, betaine biosynthesis, polyamines biosynthesis, sulfate-sulfur assimilation pathway as well as type II toxin-antitoxin (TA) system, quorum sensing (QS) system, and cellulose production could play important roles in the resilience of KMC after exposure to Mars-like stress. Our findings show the potential mechanisms through which Komagataeibacter tolerates the extraterrestrial stress and will help to understand minimal microbial composition of KMC for space travelers.

Liquid-crystalline ordering in bacterial cellulose produced by Gluconacetobaсter hansenii on glucose-containing media.

This research is dedicated to the studies of the microscale morphology of bacterial cellulose (BC) obtained by means of static cultivation of Gluconacetobacter hansenii GH-1/2008. We found that the microscale morphology depended on the BC production rate that was varied by using different glucose concentrations in the cultivation medium. It was revealed that at higher production rates, BC fibrils were aligned in a liquid-crystalline-like (LC-like) order. The observed helical alignment was always left-handed. The half-periods of the helix varied from 50 µm to 150 µm depending on the cultivation conditions. The mechanical and water absorption properties of the obtained BC pellicles were measured. The former correlated mainly with the density of the samples; the latter were the best for films with layered structure, where the BC had segregated into fleece sheets separated by gaps with low density of fibrils.

Melanoma growth in non-chemically modified translucid bacterial nanocellulose hollow and compartimentalized spheres.

BACKGROUND: Bacterial nanocellulose (BNC) has been used as cell support in numerous tissue engineering studies. Its use can be explained based on the fact its structure allows the creation of a required microenvironment for an ideal material, which supports 3D cell culture. Its structure and interconnected pores lead to animal cells adhesion and proliferation, also allowing oxygen and nutrients transportation. METHODS: We developed a new methodology to produce spherical platforms synthesized by Komagataebacter hansenii (ATCC 23769) under dynamic culture conditions in minimal medium. The chemical composition and physical properties of the platforms were evaluated. Then, human melanoma cells (SK-MEL-28) were encapsulated into the platforms and evaluated by metabolic activity, morphology and their ability on adhering to the Hollow Translucid BNC Spheres (BNC-TS-H) and Compartmentalized Translucid BNC Spheres (BNC-TS-C) up to 3 days. RESULTS: BNC-TS-H and BNC-TS-C platforms were produced as translucid spheroid platforms with distinct microenvironment under dynamic fermentation. The chemical and physical characterizations confirmed the platforms composition as BNC. The produced internal microenvironments in spherical platforms are relevant to determine tumor cell fate. In the first 12 h of culture, cells could adhere to nanocellulose microfibers assuming their typical tumorous phenotype in 72 h of culture. CONCLUSION: The dynamic fermentation in minimal medium produced distinct microstructured platforms of BNC-TS-H and BNC-TS-C. The platforms microstructure resulted in microenvironments that enabled distinct cell-cell and cell-matrix interactions. This behavior suggests several applications in tissue engineering. GENERAL SIGNIFICANCE: The method produced translucid BNC sphere platforms with distinct microenvironments for 3D cell culture.

A turning point in the bacterial nanocellulose production employing low doses of gamma radiation.

In the recent years, huge efforts have been conducted to conceive a cost-effective production process of the bacterial nanocellulose (BNC), thanks to its marvelous properties and broadening applications. Herein, we unveiled the impact of gamma irradiation on the BNC yield by a novel bacterial strain Komagataeibacter hansenii KO28 which was exposed to different irradiation doses via a designed scheme, where the productivity and the structural properties of the BNC were inspected. After incubation for 240 h, the highest BNC yield was perceived from the culture treated twice with 0.5 kGy, recording about 475% higher than the control culture. Furthermore, almost 92% of its BNC yield emerged in the first six days. The physicochemical characteristics of the BNCs were investigated adopting scanning electron microscope (SEM), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and Fourier transform infrared (FTIR). Additionally, the water holding capacity, water release rate, surface area (BET), and mechanical properties were configured for the BNC generated from the control and the irradiated cultures. As a whole, there were no significant variations in the properties of the BNC produced by the irradiated cultures versus the control, proposing the strain irradiation as a valuable, facile, and cheap route to augment the BNC yield.

Photonic Cellulose Films with Vivid Structural Colors: Fabrication and Selectively Chemical Response.

Recent advances in structural-color cellulose nanocrystal (CNC) materials have been made toward chemical sensing applications; however, such materials lack sufficient color chroma for naked-eye observation, and their selective recognition to given chemicals as well as the corresponding mechanism has rarely been reported. Here, a dopamine-infiltration and post-polymerization approach is proposed to construct vivid structural-color composite films. The chiral nematic structure of CNC enables the structural coloration, while the strong light absorption of the polymeric co-phase, polydopamine (PDA) enhances the color chroma and visibility. By controlling the PDA amount, the composite films can detect organic solvents quantitatively and selectively via visible color changes. From the viewpoint of the compatibility and similitude principle, notably, a critical solubility parameter distance (R0) between PDA and "active" solvents is defined with a three-dimensional Hansen solubility sphere; this well constructs a rule for the sensing selectivity of the chemochromic composite films. The findings pave the foundation for the design of colorimetric sensors with specifically testing objects.

Comparative Analysis of Bacterial Cellulose Membranes Synthesized by Chosen Komagataeibacter Strains and Their Application Potential.

This article presents a comparative analysis of bacterial cellulose membranes synthesized by several strains of the Komagataeibacter genus in terms of their specific physical, physico-chemical, and mechanical properties. Herein, the aim was to choose the most suitable microorganisms producing cellulosic materials with the greatest potential for the fabrication of bio-inspired nanocomposites. The selection was based on three main steps, starting from the evaluation of BNC biosynthetic efficiency with and without the addition of ethanol, followed by the assessment of mechanical breaking strength, and the physical parameters (compactness, structural integrity, appearance, and thickness) of the obtained biological materials. Ultimately, based on the performed screening procedure, three efficiently growing strains (K. hansenii H3 (6Et), K. rhaeticus K4 (8Et), and Komagataeibacter sp. isolated from balsamic vinegar (12Et)) were chosen for further modifications, enabling additional cellulose functionalization. Here, supplementation of the growth medium with five representative polymeric compounds (citrus/apple pectin, wheat starch, polyvinyl alcohol, polyethylene glycol) led to significant changes in BNC properties, especially dye loading abilities, mechanical strength, and water adsorption/retention capacities. The resulting nanocomposites can be potentially useful in various fields of medicine and industry, and in the future, they may become a practical and cost-effective competitor against commercial biomaterials currently available on the market.

The Roles of the Various Cellulose Biosynthesis Operons in Komagataeibacter hansenii ATCC 23769.

Cellulose is the most abundant biopolymer on earth and offers versatile applicability in biotechnology. Bacterial cellulose, especially, is an attractive material because it represents pure microcrystalline cellulose. The cellulose synthase complex of acetic acid bacteria serves as a model for general studies on (bacterial) cellulose synthesis. The genome of Komagataeibacter hansenii ATCC 23769 encodes three cellulose synthase (CS) operons of different sizes and gene compositions. This implies the question of which role each of the three CS-encoding operons, bcsAB1, bcsAB2, and bcsAB3, plays in overall cellulose synthesis. Therefore, we constructed markerless deletions in K. hansenii ATCC 23769, yielding mutant strains that expressed only one of the three CSs. Apparently, BcsAB1 is the only CS that produces fibers of crystalline cellulose. The markerless deletion of bcsAB1 resulted in a nonfiber phenotype in scanning electron microscopy analysis. Expression of the other CSs resulted in a different, nonfibrous extracellular polymeric substance (nfEPS) structure wrapping the cells, which is proposed to contain acetylated cellulose. Transcription analysis revealed that all CSs were expressed continuously and that bcsAB2 showed a higher transcription level than bcsAB1. Moreover, we were able to link the expression of diguanylate cyclase B (dgcB) to cellulose production. IMPORTANCE Acetic acid bacteria form a massive biofilm called "mother of vinegar," which is built of cellulose fibers. Bacterial cellulose is an appealing biomaterial with manifold applications in biomedicine and biotechnology. Because most cellulose-producing acetic acid bacteria express several cellulose synthase operons, a deeper understanding of their contribution to the synthesis of modified forms of cellulose fibers within a natural biofilm is of special interest. For the first time, we were able to identify the contribution of each of the three cellulose synthases to cellulose formation in Komagataeibacter hansenii ATCC 23769 after a chromosomal clean deletion. Moreover, we were able to depict their roles in spatial composition of the biofilm. These findings might be applicable in the future for naturally modified biomaterials with novel properties.

Effects of pullulan additive and co-culture of Aureobasidium pullulans on bacterial cellulose produced by Komagataeibacter hansenii.

Bacterial cellulose (BC) exhibits a unique combination of porosity, tensile strength, reticulated crystal structure and biocompatibility useful for many applications in the food, biomedical and other industries. Polysaccharide addition has been shown to improve the production and the mechanical properties of BC nanocomposites. This study examined the effect of pullulan on BC fermentation as well as the co-culturing of the BC producer with Aureobasidium pullulans, a fungal strain that produces pullulan as an exopolysaccharide. Results showed that a 1% pullulan addition improved Young's modulus of BC pellicles for sixfold. Addition of pullulan at 1.5% and 2% levels could increase the BC production from 0.447 to 0.814 and 1.997 g/L, respectively. The co-culture fermentation demonstrated a mixed effect on the aggregation and bundling of BC while resulting in a significant improvement in mechanical properties. The study provided a polysaccharide additive and a novel fermentation method to produce BC with improved properties.

Modified montmorillonite-bacterial cellulose composites as a novel dressing system for pressure injury.

The main objective of this study was to investigate the effects of bacterial cellulose hydrogel (BCH) incorporated into montmorillonite (MMT) and its underlying mechanisms of action on a skin wound healing mouse model following pressure injury model. Komagataeibacter hansenii was used to obtain 5 cm in diameter and 0.8 mm of thickness circular bacterial cellulose (BC) sheets, which were incorporated with MMT by deposition ex-site using a 0.1% MMT suspension (100 rpm for 24 h at 28 °C). Afterward, Fourier Transform Infrared Spectroscopy (FT-IR) and Scanning Electron Microscopy (SEM) were used to characterize the bacterial cellulose hydrogel incorporated into montmorillonite (BCH-MMT). The pressure injury model was assessed by macroscopic and histological analysis in male Swiss mice. Both, BC and BCH-MMT, showed a typical FTIR spectrum of cellulosic substrates with pronounces bands around 3344, 2920, 1637, and 1041 cm-1 while microparticles of MMT dispersed uniformly throughout BC were revealed by SEM photographs. Animals treated with BCH-MMT showed significant healing of pressure ulcers as demonstrated by reduced area of redness and spontaneous hyperalgesia, lower amounts of in-site inflammatory cells (to the same level as the positive control Dersani®) and ultimately, complete epidermis re-epithelialization and tissue regeneration. Altogether, these findings suggest that a modified BCH-MMT film could serve as scaffolding for skin tissue engineering and potentially as a novel dressing material for pressure injury.

Regioselective Protection and Deprotection of Nanocellulose Molecular Design Architecture: Robust Platform for Multifunctional Applications.

Regioselectively substituted nanocellulose was synthesized by protecting the primary hydroxyl group. Herein, we took advantage of the different reactivities of primary and secondary hydroxyl groups to graft large capping structures. This study mainly focuses on regioselective installation of trityl protecting groups on nanocellulose chains. The elemental analysis and nuclear magnetic resonance spectroscopy of regioselectively substituted nanofibrillated cellulose (NFC) suggested that the trityl group was successfully grafted in the primary hydroxyl group with a degree of substitution of nearly 1. Hansen solubility parameters were employed, and the binary system composed of an ionic liquid and pyridine as a base was revealed to be the optimum condition for regioselective functionalization of nanocellulose. Interestingly, the dissolution of NFC in the ionic liquid and the subsequent deprotection process of NFC substrates hardly affected the crystalline structure of NFC (3.6% decrease in crystallinity). This method may provide endless possibilities for the design of advanced engineered nanomaterials with multiple functionalities. We envisage that this protection/deprotection approach may lead to a bright future for the fabrication of multifunctional devices based on nanocellulose.

Biofabrication of a Hyaluronan/Bacterial Cellulose Composite Nanofibril by Secretion from Engineered Gluconacetobacter.

Naturally occurring polysaccharides, such as cellulose, hemicellulose, and chitin, have roles in plant skeletons and/or related properties in living organisms. Their hierarchically regulated production systems show potential for designing nanocomposite fabrication using engineered microorganisms. This study has demonstrated that genetically engineered Gluconacetobacter hansenii (G. hansenii) individual cells can fabricate naturally composited nanofibrils by simultaneous production of hyaluronan (HA) and bacterial cellulose (BC). The cells were manipulated to contain hyaluronan synthase and UDP-glucose dehydrogenase genes, which are essential for HA biosynthesis. Fluorescence microscopic observations indicated the production of composited nanofibrils and suggested that HA secretion was associated with the cellulose secretory pathway in G. hansenii. The gel-like nanocomposite materials produced by the engineered G. hansenii exhibited superior properties compared with conventional in situ nanocomposites. This genetic engineering approach facilitates the use of G. hansenii for designing integrated cellulose-based nanomaterials.

The influence of alkaline treatment on the mechanical and structural properties of bacterial cellulose.

The unique mechanical properties of hydrated bacterial cellulose make it suitable for biomedical applications. This study evaluates the effect of concentrated sodium hydroxide treatment on the structural and mechanical properties of bacterial cellulose hydrogels using rheological, tensile, and compression tests combined with mathematical modelling. Bacterial cellulose hydrogels show a concentration-dependent and irreversible reduction in shear moduli, compression, and tensile strength after alkaline treatment. Applying a poroelastic biphasic model to through-thickness compressive stress-relaxation tests showed the alkaline treatment to induce no significant change in axial compression, an effect was observed in the radial direction, potentially due to the escape of water from within the hydrogel. Scanning electron microscopy showed a more porous structure of bacterial cellulose. These results show how concentration-dependent alkaline treatment induces selective weakening of intramolecular interactions between cellulose fibres, allowing the opportunity to precisely tune the mechanical properties for specific biomedical application, e.g., faster-degradable materials.

Hybrid Living Capsules Autonomously Produced by Engineered Bacteria.

Bacterial cellulose (BC) has excellent material properties and can be produced sustainably through simple bacterial culture, but BC-producing bacteria lack the extensive genetic toolkits of model organisms such as Escherichia coli (E. coli). Here, a simple approach is reported for producing highly programmable BC materials through incorporation of engineered E. coli. The acetic acid bacterium Gluconacetobacter hansenii is cocultured with engineered E. coli in droplets of glucose-rich media to produce robust cellulose capsules, which are then colonized by the E. coli upon transfer to selective lysogeny broth media. It is shown that the encapsulated E. coli can produce engineered protein nanofibers within the cellulose matrix, yielding hybrid capsules capable of sequestering specific biomolecules from the environment and enzymatic catalysis. Furthermore, capsules are produced which can alter their own bulk physical properties through enzyme-induced biomineralization. This novel system uses a simple fabrication process, based on the autonomous activity of two bacteria, to significantly expand the functionality of BC-based living materials.