Engineering PTS-based glucose metabolism for efficient biosynthesis of bacterial cellulose by Komagataeibacter xylinus.

Bacterial cellulose (BC) is a renewable biomaterial that has attracted significant attention due to its excellent properties and wide applications. Komagataeibacter xylinus CGMCC 2955 is an important BC-producing strain. It primarily produces BC from glucose while simultaneously generating gluconic acid as a by-product, which acidifies the medium and inhibits BC synthesis. To enhance glucose uptake and BC synthesis, we reconstructed the phosphoenolpyruvate-dependent glucose phosphotransferase system (PTSGlc) and strengthened glycolysis by introducing heterologous genes, resulting in a recombinant strain (GX08PTS03; Δgcd::ptsHIcrrE. coli::ptsGE. coli::pfkAE. coli). Strain GX08PTS03 efficiently utilized glucose for BC production without accumulating gluconic acid. Subsequently, the fermentation process was systematically optimized. Under optimal conditions, strain GX08PTS03 produced 7.74 g/L of BC after 6 days of static fermentation, with a BC yield of 0.39 g/g glucose, which were 87.41 % and 77.27 % higher than those of the wild-type strain, respectively. The BC produced by strain GX08PTS03 exhibited a longer fiber diameter along with a lower porosity, significantly higher solid content, crystallinity, tensile strength, and Young's modulus. This study is novel in reporting that the engineered PTSGlc-based glucose metabolism could effectively enhance the production and properties of BC, providing a future outlook for the biopolymer industry.

Directed evolution of material-producing microorganisms.

Nature is home to a variety of microorganisms that create materials under environmentally friendly conditions. While this offers an attractive approach for sustainable manufacturing, the production of materials by native microorganisms is usually slow and synthetic biology tools to engineer faster microorganisms are only available when prior knowledge of genotype-phenotype links is available. Here, we utilize a high-throughput directed evolution platform to enhance the fitness of whole microorganisms under selection pressure and identify genetic pathways to enhance the material production capabilities of native species. Using Komagataeibacter sucrofermentans as a model cellulose-producing microorganism, we show that our droplet-based microfluidic platform enables the directed evolution of these bacteria toward a small number of cellulose overproducers from an initial pool of 40,000 random mutants. Sequencing of the evolved strains reveals an unexpected link between the cellulose-forming ability of the bacteria and a gene encoding a protease complex responsible for protein turnover in the cell. The ability to enhance the fitness of microorganisms toward a specific phenotype and to unravel genotype-phenotype links makes this high-throughput directed evolution platform a promising tool for the development of new strains for the sustainable manufacturing of materials.

Enhanced bacterial cellulose production by Komagataeibacter species and Hibiscus sabdariffa herbal tea.

This study proposed Hibiscus sabdariffa as a novel substrate for BC production with Komagataeibacter species and their consortia. K. intermedius is found as the most efficient producer (5.98 g/L BC, 3.56 × 10-2 g-1 h-1 productivity rate) following K. maltaceti (4.44 g/L BC, 2.64 × 10-2 g-1 h-1 productivity rate) and K. nataicola (3.67 g/L BC, 2.18 × 10-2 g-1 h-1 productivity rate). Whereas agitation increased BC production with K. nataicola (1.22-fold, 4.49 g/L BC), K. maltaceti (1.24-fold, 5.52 g/L BC) and K. intermedius (1.27-fold, 7.63 g/L BC), irregular shaped BC was obtained. This could be a novel result as Komagataeibacter consortia increased BC production by 1.17-2.01-fold compared to monocultures resulting as 8.11 g/L BC through the co-cultivation of K. maltaceti-K. intermedius. Maximum increase was found to be 1.75-fold (1.79-fold WHC), occurring with monoculture of K. maltaceti, while 1.94-fold (1.26-fold WHC) with K. maltaceti-K. intermedius consortium when H. sabdariffa-based media compared Hestrin-Schramm media. Based on these results, this could be a novel result as H. sabdariffa-based media may replace the use of HS media in BC production by means of a bioactive content-rich plant and obtaining 3-D ultrafine porous structure with high thermal resistant (∼460-500 °C) BC with mono and co-cultivation of Komagataeibacter species to be used in industrial area.

A novel cost-effective methodology for the screening of nanocellulose producing micro-organisms.

In this paper, the work has been done to develop a cost-effective methodology, for the isolation of the potential producer of bacterial nanocellulose. No report is available in the literature, on the use of gram flour and table sugar for the screening of nanocellulose-producing isolates. Since commercially used, Hestrin-Schramm medium is expensive for the isolation of nanocellulose-producing micro-organisms, the possibility of using gram flour-table sugar medium was investigated in this work. Qualitative screening of micro-organisms was done using cost-effective medium, i.e., gram flour-table sugar medium. Qualitative analysis of various nanocellulose-producing bacteria depicted that cellulose layer production occurred on both HS medium and gram flour-table sugar medium. The yield of nanocellulose was also better on air-liquid surface in case of gram flour-table sugar medium as compared to HS medium. 16S rRNA was used for molecular characterization of bacterial strain and the best nanocellulose producer was identified as Novacetimonas hansenii BMK-3_NC240423 (isolated from rotten banana). FTIR and FE-SEM studies of nanocellulose pellicle produced on HS medium and gram flour-table sugar medium demonstrated equivalent structural, morphological, and chemical properties. The cost of newly designed medium (0.01967 $/L) is nearly 90 times lower than the Hestrin-Schramm medium (1.748 $/L), which makes the screening of nanocellulose producers very cost-effective. A strategy of using gram flour extract-table sugar medium for the screening of nanocellulose-producing micro-organisms is a novel approach, which will drastically reduce the screening associated cost of cellulose-producing micro-organisms and also motivate the researchers/industries for comprehensive screening programme for getting high cellulose-producing microbes.

Bacterial valorization of agricultural-waste into a nano-sized cellulosic matrix for mitigating emerging pharmaceutical pollutants: An eco-benign approach.

For Bacterial Nanocellulose (BNC) production, standard methods are well-established, but there is a pressing need to explore cost-effective alternatives for BNC commercialization. This study investigates the feasibility of using syrup prepared from maize stalk as a valuable nutrient and sustainable carbon source for BNC production. Our study achieved a remarkable BNC production yield of 19.457 g L-1 by utilizing Komagataeibacter saccharivorans NUWB1 in combination with components from the Hestrin-Schramm (HS) medium. Physicochemical properties revealed that the obtained BNC exhibited a crystallinity index of 60.5 %, tensile strength of 43.5 MPa along with enhanced thermostability reaching up to 360 °C. N2 adsorption-desorption isotherm of the BNC displayed characteristics of type IV, indicating the presence of a mesoporous structure. The produced BNC underwent thorough investigation, focusing on its efficacy in addressing environmental concerns, particularly in removing emerging pharmaceutical pollutants like Metformin and Paracetamol. Remarkably, the BNC exhibited strong adsorption capabilities, aligning with the Langmuir isotherm and pseudo-second-order model. Thermodynamic analysis confirmed a spontaneous and endothermic adsorption process. Furthermore, the BNC showed potential for regeneration, enabling up to five recycling cycles. Cytotoxicity and oxidative stress assays validated the biocompatibility of BNC. Lastly, the BNC films displayed an impressive 88.73 % biodegradation within 21 days.

Evaluation of hydrolyzed cheese whey medium for enhanced bacterial cellulose production by Komagataeibacter rhaeticus MSCL 1463.

Industrial production of bacterial cellulose (BC) remains challenging due to significant production costs, including the choice of appropriate growth media. This research focuses on optimization of cheese whey (CW) based media for enhanced production of BC. Two modifications were made for CW medium for BC production with Komagataeibacter rhaeticus MSCL 1463. BC production in a medium of enzymatically hydrolyzed CW (final concentration of monosaccharides: glucose 0.13 g L-1, galactose 1.24 g L-1) was significantly enhanced, achieving a yield of 4.95 ± 0.25 g L-1, which markedly surpasses the yields obtained with the standard Hestrin-Schramm (HS) medium containing 20 g L-1 glucose and acid-hydrolyzed CW (final concentration of monosaccharides: glucose 1.15 g L-1, galactose 2.01 g L-1), which yielded 3.29 ± 0.12 g L-1 and 1.01 ± 0.14 g L-1, respectively. We explored the synergistic effects of combining CW with various agricultural by-products (corn steep liquor (CSL), apple juice, and sugar beet molasses). Notably, the supplementation with 15% corn steep liquor significantly enhanced BC productivity, achieving 6.97 ± 0.17 g L-1. A comprehensive analysis of the BC's physical and mechanical properties indicated significant alterations in fiber diameter (62-167 nm), crystallinity index (71.1-85.9%), and specific strength (35-82 MPa × cm3 g-1), as well as changes in the density (1.1-1.4 g cm-3). Hydrolyzed CW medium supplemented by CSL could be used for effective production of BC.

[Knockdown of motility-related genes of Komagataeibacter xylinus and its effect on bacterial cellulose synthesis].

Bacterial cellulose (BC) is a biopolymer synthesized by bacteria, which possess excellent characteristics such as high water holding capacity, high crystallinity, and high purity. It is widely used in food, medical, cosmetics, and functional films. Komagataeibacter xylinus is a model strain used in BC synthesis research. In bacteria, motility-related genes are associated with BC synthesis, whereas in Komagataeibacter xylinus CGMCC 2955, the functions of motility-related genes and their effects on BC synthesis are not known. To address this gap, we used the λ Red recombinant system to individually knock out motA, motB, and mot2A respectively, and constructed the knockout strains K. x-ΔmotA, K. x-ΔmotB, and K. x-Δmot2A. Additionally, both motA and motB were disrupted to construct the K. x-ΔmotAB mutant. The results demonstrated that knockout strain K. x-ΔmotAB exhibited the highest BC yield, reaching (5.05±0.26) g/L, which represented an increase of approximately 24% compared to wild-type strains. Furthermore, the BC synthesized by this strain exhibited the lowest porosity, 54.35%, and displayed superior mechanical properties with a Young's modulus of up to 5.21 GPa. As knocking out motA and motB genes in K. xylinus CGMCC 2955 did not reduce BC yield; instead, it promoted BC synthesis. Consequently, this research further deepened our understanding of the relationship between motility and BC synthesis in acetic acid bacteria. The knockouts of motA and motB genes resulted in reduced BC porosity and improved mechanical properties, provides a reference for BC synthesis and membrane structure regulation modification.

Production and characterization of bacterial cellulose by Rhizobium sp. isolated from bean root.

Bacterial cellulose (BC) is a natural polymer renowned for its unique physicochemical and mechanical attributes, including notable water-holding capacity, crystallinity, and a pristine fiber network structure. While BC has broad applications spanning agriculture, industry, and medicine, its industrial utilization is hindered by production costs and yield limitations. In this study, Rhizobium sp. was isolated from bean roots and systematically assessed for BC synthesis under optimal conditions, with a comparative analysis against BC produced by Komagataeibacter hansenii. The study revealed that Rhizobium sp. exhibited optimal BC synthesis when supplied with a 1.5% glucose carbon source and a 0.15% yeast extract nitrogen source. Under static conditions at 30 °C and pH 6.5, the most favorable conditions for growth and BC production (2.5 g/L) were identified. Modifications were introduced using nisin to enhance BC properties, and the resulting BC-nisin composites were comprehensively characterized through various techniques, including FE-SEM, FTIR, porosity, swelling, filtration, and antibacterial activity assessments. The results demonstrated that BC produced by Rhizobium sp. displayed properties comparable to K. hansenii-produced BC. Furthermore, the BC-nisin composites exhibited remarkable inhibitory activity against Escherichia coli and Pseudomonas aeruginosa. This study contributes valuable insights into BC's production, modification, and characterization utilizing Rhizobium sp., highlighting the exceptional properties that render it efficacious across diverse applications.

Microorganisms and bacterial cellulose stability of Kombucha under different manufacture and storage conditions.

It is crucial to clarify the stability of Kombucha in the manufacture and storage stages due to the extensive study on the fermented products of Kombucha and the increase in the use of bacterial cellulose (BC). This study aimed to evaluate the stability of Kombucha in different manufacturing and storage temperatures within a certain time period. The stability of microorganisms and BC in Kombucha was investigated through regular replacement with the tea media at 28 and 25°C for manufacture, and the storage temperature of Kombucha was at 25, 4, and -20°C. Morphological observations of the BC in Kombucha ended at 28 and 25°C for manufacture and storage were performed using atomic force microscopy (AFM) before inoculation. The viable cell counts and AFM results showed that the stability of Kombucha during manufacture was better at 28°C than at 25°C, with higher microbial viability and BC productivity in the former at the time of manufacture, whereas 25°C was more favorable for the stability of Kombucha during storage. At the same temperature of 25°C, the manufacturing practice improved the microbial viability and BC stability compared with storage; the pH value of Kombucha was lower, and the dry weight of BC was higher during storage compared with manufacture. The maximum BC water holding capacity (97.16%) was maintained by storage at 4°C on day 63, and the maximum BC swelling rate (56.92%) was observed after storage at -20°C on day 7. The research was conducted to provide reference information for applying Kombucha and its BC in food and development in other industries.

Bacterial nanocellulose: A versatile biopolymer production using a cost-effective wooden disc based rotary reactor.

Bacterial nanocellulose (BNC) has various unique qualities, including high mechanical strength, crystallinity, and high water-holding capacity, which makes it appropriate for a wide range of industrial applications. But its lower yield coupled with its high production cost creates a barrier to its usage. In this study, we have demonstrated the better yield of BNC from an indigenous strain Komagataeibacter rhaeticus MCC-0157 using a rotary disc bioreactor (RDB) having a wooden disc. The RDB was optimized based on the type of disc material, distance between the disc, and rotation speed to get the highest yield of 13.0 g/L dry material using Hestrin-Schramm (H-S) medium. Further, the bioreactor was compared for the BNC production using reported medium, which is used for static condition; the RDB showed up to fivefold increase in comparison with the static condition reported. Komagataeibacter rhaeticus MCC-0157 was previously reported to be one of the highest BNC producing stains, with 8.37 g/L of dry yield in static condition in 15 days incubation. The designed RDB demonstrated 13.0 g/L dry yield of BNC in just 5 days. Other characteristics of BNC remain same as compared with static BNC production, although the difference in the crystallinity index was observed in RDB (84.44%) in comparison with static (89.74%). For the first time, wooden disc was used for rotary bioreactor approach, which demonstrated higher yield of BNC in lesser time and can be further used for sustainable production of BNC at the industrial level.

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.

An evaluation of fusion partner proteins for paratransgenesis in Asaia bogorensis.

Mosquitoes transmit many pathogens responsible for human diseases, such as malaria which is caused by parasites in the genus Plasmodium. Current strategies to control vector-transmitted diseases are increasingly undermined by mosquito and pathogen resistance, so additional methods of control are required. Paratransgenesis is a method whereby symbiotic bacteria are genetically modified to affect the mosquito's phenotype by engineering them to deliver effector molecules into the midgut to kill parasites. One paratransgenesis candidate is Asaia bogorensis, a Gram-negative bacterium colonizing the midgut, ovaries, and salivary glands of Anopheles sp. mosquitoes. Previously, engineered Asaia strains using native signals to drive the release of the antimicrobial peptide, scorpine, fused to alkaline phosphatase were successful in significantly suppressing the number of oocysts formed after a blood meal containing P. berghei. However, these strains saw high fitness costs associated with the production of the recombinant protein. Here, we report evaluation of five different partner proteins fused to scorpine that were evaluated for effects on the growth and fitness of the transgenic bacteria. Three of the new partner proteins resulted in significant levels of protein released from the Asaia bacterium while also significantly reducing the prevalence of mosquitoes infected with P. berghei. Two partners performed as well as the previously tested Asaia strain that used alkaline phosphatase in the fitness analyses, but neither exceeded it. It may be that there is a maximum level of fitness and parasite inhibition that can be achieved with scorpine being driven constitutively, and that use of a Plasmodium specific effector molecule in place of scorpine would help to mitigate the stress on the symbionts.

Characterization of the Putative Acylated Cellulose Synthase Operon in Komagataeibacter xylinus E25.

Bacterial cellulose is a natural polymer with an expanding array of applications. Because of this, the main cellulose producers of the Komagataeibacter genus have been extensively studied with the aim to increase its synthesis or to customize its physicochemical features. Up to now, the genetic studies in Komagataeibacter have focused on the first cellulose synthase operon (bcsI) encoding the main enzyme complex. However, the role of other accessory cellulose operons has been understudied. Here we aimed to fill this gap by performing a detailed analysis of the second cellulose synthase operon (bcsII), which is putatively linked with cellulose acylation. In this study we harnessed the genome sequence, gene expression and protein structure information of K. xylinus E25 and other Komagataeibacter species to discuss the probable features of bcsII and the biochemical function of its main protein products. The results of our study support the previous hypothesis that bcsII is involved in the synthesis of the acylated polymer and expand it by presenting the evidence that it may also function in the regulation of its attachment to the cell surface and to the crystalline cellulose fibers.

Bacterial Cellulose Production from agricultural Residues by two Komagataeibacter sp. Strains.

Agricultural residues are constantly increasing with increased farming processes, and improper disposal is detrimental to the environment. Majority of these waste residues are rich in lignocellulose, which makes them suitable substrate for bacterial fermentation in the production of value-added products. In this study, bacterial cellulose (BC), a purer and better form of cellulose, was produced by two Komagataeibacter sp. isolated from rotten banana and kombucha drink using corncob (CC) and sugarcane bagasse (SCB) enzymatic hydrolyzate, under different fermentation conditions, that is, static, continuous, and intermittent agitation. The physicochemical and mechanical properties of the BC films were then investigated by Fourier Transformed Infrared Spectroscopy (FTIR), Thermogravimetry analysis, Field Emission Scanning Electron Microscopy (FE-SEM), and Dynamic mechanical analysis. Agitation gave a higher BC yield, with Komagataeibacter sp. CCUG73629 producing BC from CC with a dry weight of 1.6 g/L and 1.4 g/L under continuous and intermittent agitation, respectively, compared with that of 0.9 g/L in HS medium. While BC yield of dry weight up to 1.2 g/L was obtained from SCB by Komagataeibacter sp. CCUG73630 under continuous agitation compared to that of 0.3 g/L in HS medium. FTIR analysis showed BC bands associated with cellulose I, with high thermal stability. The FE-SEM analysis showed that BC fibers were highly ordered and densely packed. Although the BC produced by both strains showed similar physicochemical and morphological properties, the BC produced by the Komagataeibacter sp. CCUG73630 in CC under intermittent agitation had the best modulus of elasticity, 10.8 GPa and tensile strength, 70.9 MPa.

One-Step Biosynthesis of Soft Magnetic Bacterial Cellulose Spheres with Localized Nanoparticle Functionalization.

Actuated structures are becoming relevant in medical fields; however, they call for flexible/soft-base materials that comply with biological tissues and can be synthesized in simple fabrication steps. In this work, we extend the palette of techniques to afford soft, actuable spherical structures taking advantage of the biosynthesis process of bacterial cellulose. Bacterial cellulose spheres (BCS) with localized magnetic nanoparticles (NPs) have been biosynthesized using two different one-pot processes: in agitation and on hydrophobic surface-supported static culture, achieving core-shell or hollow spheres, respectively. Magnetic actuability is conferred by superparamagnetic iron oxide NPs (SPIONs), and their location within the structure was finely tuned with high precision. The size, structure, flexibility and magnetic response of the spheres have been characterized. In addition, the versatility of the methodology allows us to produce actuated spherical structures adding other NPs (Au and Pt) in specific locations, creating Janus structures. The combination of Pt NPs and SPIONs provides moving composite structures driven both by a magnetic field and a H2O2 oxidation reaction. Janus Pt/SPIONs increased by five times the directionality and movement of these structures in comparison to the controls.

Bacterial cellulose adhesive composites for oral cavity applications.

Topical approaches to oral diseases require frequent dosing due to limited retention time. A mucoadhesive drug delivery platform with extended soft tissue adhesion capability of up to 7 days is proposed for on-site management of oral wound. Bacterial cellulose (BC) and photoactivated carbene-based bioadhesives (PDz) are combined to yield flexible film platform for interfacing soft tissues in dynamic, wet environments. Structure-activity relationships evaluate UV dose and hydration state with respect to adhesive strength on soft tissue mimics. The bioadhesive composite has an adhesion strength ranging from 7 to 17 kPa and duration exceeding 48 h in wet conditions under sustained shear forces, while other mucoadhesives based on hydrophilic macromolecules exhibit adhesion strength of 0.5-5 kPa and last only a few hours. The work highlights the first evaluation of BC composites for mucoadhesive treatments in the buccal cavity.

Overabundance of Asaia and Serratia Bacteria Is Associated with Deltamethrin Insecticide Susceptibility in Anopheles coluzzii from Agboville, Côte d'Ivoire.

Insecticide resistance among mosquito species is now a pervasive phenomenon that threatens to jeopardize global malaria vector control efforts. Evidence of links between the mosquito microbiota and insecticide resistance is emerging, with significant enrichment of insecticide degrading bacteria and enzymes in resistant populations. Using 16S rRNA amplicon sequencing, we characterized and compared the microbiota of Anopheles coluzzii in relation to their deltamethrin resistance and exposure profiles. Comparisons between 2- and 3-day-old deltamethrin-resistant and -susceptible mosquitoes demonstrated significant differences in microbiota diversity. Ochrobactrum, Lysinibacillus, and Stenotrophomonas genera, each of which comprised insecticide-degrading species, were significantly enriched in resistant mosquitoes. Susceptible mosquitoes had a significant reduction in alpha diversity compared to resistant individuals, with Asaia and Serratia dominating microbial profiles. There was no significant difference in deltamethrin-exposed and -unexposed 5- to 6-day-old individuals, suggesting that insecticide exposure had minimal impact on microbial composition. Serratia and Asaia were also dominant in 5- to 6-day-old mosquitoes, which had reduced microbial diversity compared to 2- to 3-day-old mosquitoes. Our findings revealed significant alterations of Anopheles coluzzii microbiota associated with deltamethrin resistance, highlighting the potential for identification of novel microbial markers for insecticide resistance surveillance. qPCR detection of Serratia and Asaia was consistent with 16S rRNA sequencing, suggesting that population-level field screening of bacterial microbiota may be feasibly integrated into wider resistance monitoring, if reliable and reproducible markers associated with phenotype can be identified. IMPORTANCE Control of insecticide-resistant vector populations remains a significant challenge to global malaria control and while substantial progress has been made elucidating key target site mutations, overexpressed detoxification enzymes and alternate gene families, the contribution of the mosquito microbiota to phenotypic insecticide resistance has been largely overlooked. We focused on determining the effects of deltamethrin resistance intensity on Anopheles coluzzii microbiota and identifying any microbial taxa associated with phenotype. We demonstrated a significant reduction in microbial diversity between deltamethrin-resistant and -susceptible mosquitoes. Insecticide degrading bacterial species belonging to Ochrobactrum, Lysinibacillus, and Stenotrophomonas genera were significantly enriched in resistant mosquitoes, while Asaia and Serratia dominated microbial profiles of susceptible individuals. Our results revealed significant alterations of Anopheles coluzzii microbiota associated with deltamethrin resistance, highlighting the potential for identification of novel microbial markers for surveillance and opportunities for designing innovative control techniques to prevent the further evolution and spread of insecticide resistance.

Influence of cellulose nanocrystal addition on the production and characterization of bacterial nanocellulose.

Bacterial nanocellulose (BNC) is characterized by high purity and excellent mechanical properties; however, its production is constrained by low yield. Therefore, efforts aimed at improving its yield and material properties are imperative. This study investigated the effect of adding different concentrations (0%, 0.5%, and 1.0%) of cellulose nanocrystal (CNC) in Hestrin-Schramm modified medium on the yield and properties of BNC produced by Komagataeibacter sp. SFCB22-18. The BNC yield increased as following an increase in added CNC concentration. Also, the morphology, structure, crystallinity, thermal stability, and mechanical properties of BNC improved after CNC incorporation. A low CNC concentration (0.1%) favored mechanical strength, whereas 0.5% gave the optimum morphology, structural, and thermal stability. These results showed that modifying BNC with CNC could help increase yield and improve its properties, and thus; the potentiality of BNC in various applications would be much enhanced.

Evaluation of carbon sources from sugar industry to bacterial nanocellulose produced by Komagataeibacter xylinus.

Nanocellulose derived from microorganism is crucial bio-based products due to its unique physicochemical and mechanical properties for material science. Thus, optimizing bacterial cellulose (BNC) production is essential to widen applications and reduce production cost. Using various carbon sources derive from fruits as alternatives for synthesizing BNC could produce a low-cost BNC with comparable properties. Although Komagataeibacter xylinus grown in different natural juices, including clarified juice (CJ), sugarcane juice (SC) and coconut juice (CN) demonstrated a lower yield than that of control medium (HS), FTIR confirmed no change in chemical functional groups of BNCs. Similarly, different sugar sources have slightly effects on mechanical and thermal properties of BNC. However, the internal morphology illustrated the pore structure in oval shape for HS and CN while CJ and SC resulted in irregular pores which could lead to the highest crystallinity index value for BNC from HS compared to that from alternative media.

Bio-conversion of kitchen waste into bacterial cellulose using a new multiple carbon utilizing Komagataeibacter rhaeticus: Fermentation profiles and genome-wide analysis.

A cellulose-producing bacterium Komagataeibacter rhaeticus K15 was isolated from kombucha tea, and its metabolic pathways and cellulose synthesis operon were analyzed by genome sequencing. Different from the reported K. rhaeticus, the K15 produced little gluconic acid (2.26 g/L) when glucose was the sole carbon source and has the capacity for high cellulose production (4.76 g/L) with other carbon sources. Furthermore, six nitrogen-fixing genes were found to be responsible for the survival of K15 on a nitrogen-free medium. Based on its fermentation characteristics, K15 was cultured in a kitchen waste medium as a strategy for green and sustainable bacterial cellulose production. The SEM, XRD, and FTIR results indicated that synthesized cellulose has a mean diameter of 40-50 nm nanofiber, good crystallinity, and the same chemical structure. The K15 strain provides a highly viable alternative strategy to reduce the costs of bacterial cellulose production using agro-industrial residues as nutrient sources.