Bacterial nanocellulose (BNC) is a unique product of microbiological synthesis, having a lot of applications among which the most important is biomedicine. Objective complexities in scaling up the biosynthesis of BNC are associated with the nature of microbial producers for which BNC is not the target metabolite, therefore biosynthesis lasts long, with the BNC yield being small. Thus, the BNC scale-up problem has not yet been overcome. Here we performed biosynthesis of three scaled sheets of BNC (each having a surface area of 29,400 cm², a container volume of 441 L, and a nutrient medium volume of 260 L and characterized them. The static biosynthesis of BNC in a semisynthetic nutrient medium was scaled up using the Medusomyces gisevii Sa-12 symbiotic culture. The experiment was run in duplicate. The BNC pellicle was removed once from the nutrient medium in the first experiment and twice in the second experiment, in which case the inoculum and glucose were not additionally added to the medium. The resultant BNC sheets were characterized by scanning electron microscopy, capillary viscosimetry, infrared spectroscopy, thermomechanical and thermogravimetric analyses. When the nutrient medium was scaled up from 0.1 to 260 L, the elastic modulus of BNC samples increased tenfold and the degree of polymerization 2.5-fold. Besides, we demonstrated that scaled BNC sheets could be removed at least twice from one volume of the nutrient medium, with the yield and quality of BNC remaining the same. Consequently, the world’s largest BNC sheets 210 cm long and 140 cm wide, having a surface area of 29,400 cm² each (weighing 16.24 to 17.04 kg), have been obtained in which an adult with burns or vast wounds can easily be wrapped. The resultant sheets exhibit a typical architecture of cellulosic fibers that form a spatial 3D structure which refers to individual and extremely important characteristics of BNC. Here we thus demonstrated the scale-up of biosynthesis of BNC with improved properties, and this result was achieved by using the symbiotic culture.

One of the ways to enhance the yield of bacterial cellulose (BC) is by using dynamic aeration and different-type bioreactors because the microbial producers are strict aerobes. But in this case, the BC quality tends to worsen. Here we have combined static culture with aeration in the biosynthesis of BC by symbiotic Medusomyces gisevii Sa-12 for the first time. A new aeration method by feeding the air onto the growth medium surface is proposed herein. The culture was performed in a Binder-400 climate chamber. The study found that the air feed at a rate of 6.3 L/min allows a 25% increase in the BC yield. Moreover, this aeration mode resulted in BC samples of stable quality. The thermogravimetric and X-ray structural characteristics were retained: the crystallinity index in reflection and transmission geometries were 89% and 92%, respectively, and the allomorph Iα content was 94%. Slight decreases in the degree of polymerization (by 12.0% compared to the control―no aeration) and elastic modulus (by 12.6%) are not critical. Thus, the simple aeration by feeding the air onto the culture medium surface has turned out to be an excellent alternative to dynamic aeration. Usually, when the cultivation conditions, including the aeration ones, are changed, characteristics of the resultant BC are altered either, due to the sensitivity of individual microbial strains. In our case, the stable parameters of BC samples under variable aeration conditions are explained by the concomitant factors: the new efficient aeration method and the highly adaptive microbial producer―symbiotic Medusomyces gisevii Sa-12.

Extended cultivation with multiple removal of BC pellicles is proposed herein as a new biosynthetic process for bacterial cellulose (BC). This method enhances the BC surface area by 5–11 times per unit volume of the growth medium, improving the economic efficiency of biosynthesis. The resultant BC gel-films were thin, transparent, and congruent. The degree of polymerization (DP) and elastic modulus (EM) depended on the number of BC pellicle removals, vessel shape, and volume. The quality of BC from removals II–III to VII was better than from removal I. The process scale-up of 1:40 by volume increased DP by 1.5 times and EM by 5 times. A fact was established that the symbiotic Medusomyces gisevii Sa-12 was adaptable to exhausted growth medium: the medium was able to biosynthesize BC for 60 days, while glucose ran low at 24 days. On extended cultivation, DP and EM were found to decline by 39–64% and 57–65%, respectively. The BC gel-films obtained upon removals I–VI were successfully trialed in experimental tension-free hernioplasty.

Scaling biosynthesis of bacterial nanocellulose (BNC) allowed samples of composite paper with an increased proportion of BNC to be obtained. This work aims to study BNC samples and bleached soft- wood kraft pulp (BSKP) composite paper with a ratio of components varying across a wide range: 10:90, 30:70, 50:50, 60:40, 70:30, 90:10. The method of paper manufacturing was chosen based on the determinations of strength and deformation properties of composite samples with the BNC:BSKP ratio of 20:80. Surface application of BNT on BSKP handsheet provided for an increase in the strength values (tear resistance - by 37%, burst index - by 17%) and deformation characteristics (tension stiffness - by 66%, fracture work - by 8%, breaking length - by 4%) compared to a reference sample. The formation of composites is confirmed in all samples. Scanning electron spectroscopy revealed that paper composites comprise interlaced micro BSKP and nano BNC fibres. As the proportion of BNC in composites elevated, densification of the structure was observed due to an increased fraction of cross-linked nanosized elements. IR spectroscopy indicated the resemblance of cellulose structure in all samples. It was found that an increase in the degree of polymerisation of composite paper is directly proportional to an increase in the BNC amount in the samples. The filtering ability of composite paper samples against microorganisms in the culture liquid of the Medusomyces gisevii Sa-12 producer was studied. It should be noted that yeast retention is achieved with 70% BNC in the paper composite. The presented properties of the new material determine prospects for its use in filtering microorganisms.