Ethanol exerts a strong positive effect on the cellulose yields from the widely exploited microbial producers of the Komagataeibacter genus. Ethanol is postulated to provide an alternative energy source, enabling effective use of glucose for cellulose biosynthesis rather than for energy acquisition. In this paper, we investigate the effect of ethanol supplementation on the global gene expression profile of Komagataeibacter xylinus E25 using RNA sequencing technology (RNA-seq). We demonstrate that when ethanol is present in the culture medium, glucose metabolism is directed towards cellulose production due to the induction of genes related to UDP-glucose formation and the repression of genes involved in glycolysis and acetan biosynthesis. Transcriptional changes in the pathways of cellulose biosynthesis and c-di-GMP metabolism are also described. The transcript level profiles suggest that Schramm-Hestrin medium supplemented with ethanol promotes bacterial growth by inducing protein biosynthesis and iron uptake. We observed downregulation of genes encoding transposases of the IS 110 family which may provide one line of evidence explaining the positive effect of ethanol supplementation on the genotypic stability of K. xylinus E25. The results of this study increase knowledge and understanding of the regulatory effects imposed by ethanol on cellulose biosynthesis, providing new opportunities for directed strain improvement, scaled-up bionanocellulose production, and wider industrial exploitation of the Komagataeibacter species as bacterial cellulose producers. Electronic supplementary material The online version of this article (10.1007/s00253-019-09904-x) contains supplementary material, which is available to authorized users.
Bacterial nanocellulose (BNC) and two BNC-based composites with carboxymethyl cellulose or hydroxyethyl cellulose (BNC-CMC or BNC-HEC, respectively), were produced in situ by Komagataeibacter xylinus E25 under stationary conditions and plasticized with glycerol (ex situ modification). The BNC-CMC composite had the loosest structure (visible in SEM images) and was less crystalline (CI of 88.6%) than BNC (CI of 92.9%) and BNC-HEC (CI of 90.4%). Cellulose fibers synthesized by K. xylinus E25 in the presence of HEC were thinner in comparison to the fibers of control BNC while there was no difference in the fibers width between the BNC-CMC and control BNC. The glycerol-plasticized BNC, BNC-CMC and BNC-HEC membranes were flexible after drying, and absorbed high amounts of artificial exudate and water after rehydration. BNC-CMC treated with 2.5% v/v aqueous glycerol was characterized by the greatest free swell absorptive capacity (up to 19 g artificial exudate/g dry weight in 24 h) while the highest rehydration capacity (around 96% of the initial water content) was observed in case of BNC-CMC plasticized with 10% v/v glycerol and dehydrated. The in situ and ex situ modifications of BNC affected also the tensile strength. The highest values of tensile strength at break (around 152.2 N) and Young's modulus (around 290.3 MPa) were observed in case of the BNC-CMC composite plasticized with 2.5% v/v glycerol. The impact of plasticized BNC, BNC-CMC and BNC-HEC on the viability of HaCaT keratinocytes was also studied and found to be positive at glycerol concentrations up to 2.5% (v/v) that suggests their potential utility as wound dressings.
Featured Application: Bionanocellulose/κ-carrageenan composites developed within this study meet the specific tissue engineering requirements, including high tensile and compression strength, water holding capacity, and water retention ratio, suitable swelling properties as well as the positive effect on cell differentiation. As the key properties of these composites may be easily modified during their fabrication, the established procedure may lead to the production of customized scaffolds.Abstract: In this work, novel bacterial cellulose/κ-carrageenan (BNC/κ-Car) composites, being potential scaffolds for tissue engineering (TE), and outperforming the two polymers when used as scaffolds separately, were for the first time obtained using an in situ method, based on the stationary culture of bacteria Komagateibacter xylinus E25. The composites were compared with native BNC in terms of the morphology of fibers, chemical composition, crystallinity, tensile and compression strength, water holding capacity, water retention ratio and swelling properties. Murine chondrogenic ATDC5 cells were applied to assess the utility of the BNC/κ-Car composites as potential scaffolds. The impact of the composites on the cells viability, chondrogenic differentiation, and expression patterns of Col1α1, Col2α1, Runx2, and Sox9, which are indicative of ATDC5 chondrogenic differentiation, was determined. None of the composites obtained in this study caused the chondrocyte hypertrophy. All of them supported the differentiation of ATDC5 cells to more chondrogenic phenotype.
A new strain of bacteria producing cellulose was isolated from Kombucha and identified as Komagataeibacter hansenii, named SI1. In static conditions, the strain synthesises bacterial nanocellulose with an improved ability to stretch. In this study, utilisation of various carbon and nitrogen sources and the impact of initial pH was assessed in terms of bacterial nanocellulose yield and properties. K. hansenii SI1 produces cellulose efficiently in glycerol medium at pH 5.0–6.0 with a yield of 3.20–3.60 g/L. Glucose medium led to the synthesis of membrane characterised by a strain of 77%, which is a higher value than in the case of another Komagataeibacter species. Supplementation of medium with vitamin C results in an enhanced porosity and improves the ability of bacterial nanocellulose to stretch (up to 123%). The properties of modified membranes were studied by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction and mechanical tests. The results show that bacterial nanocellulose produced in SH medium and vitamin C-supplemented medium has unique properties (porosity, tensile strength and strain) without changing the chemical composition of cellulose. The method of production BNC with altered properties was the issue of Polish patent application no. P.431265.
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