Bacterial cellulose (BC) is a biopolymer produced by different microorganisms, but in biotechnological practice, Komagataeibacter xylinus is used. The micro- and nanofibrillar structure of BC, which forms many different-sized pores, creates prerequisites for the introduction of other polymers into it, including those synthesized by other microorganisms. The study aims to develop a cocultivation system of BC and prebiotic producers to obtain BC-based composite material with prebiotic activity. In this study, pullulan (PUL) was found to stimulate the growth of the probiotic strain Lactobacillus rhamnosus GG better than the other microbial polysaccharides gellan and xanthan. BC/PUL biocomposite with prebiotic properties was obtained by cocultivation of Komagataeibacter xylinus and Aureobasidium pullulans, BC and PUL producers respectively, on molasses medium. The inclusion of PUL in BC is proved gravimetrically by scanning electron microscopy and by Fourier transformed infrared spectroscopy. Cocultivation demonstrated a composite effect on the aggregation and binding of BC fibers, which led to a significant improvement in mechanical properties. The developed approach for “grafting” of prebiotic activity on BC allows preparation of environmentally friendly composites of better quality.

Infections of agricultural crops caused by pathogen ic fungi are among the most widespread and harmful, as they not only reduce the quantity of the harvest but also significantly deteriorate its quality. This study aims to develop unique seed-coating formulations incorporating biopolymers (polyhydroxyalkanoate and pullulan) and beneficial microorganisms for plant protection against phytopathogens. A microbial association of biocompatible endophytic bacteria has been created, including Pseudomonas flavescens D5, Bacillus aerophilus A2, Serratia proteamaculans B5, and Pseudomonas putida D7. These strains exhibited agronomically valuable properties: synthesis of the phytohormone IAA (from 45.2 to 69.2 µg mL⁻¹), antagonistic activity against Fusarium oxysporum and Fusarium solani (growth inhibition zones from 1.8 to 3.0 cm), halotolerance (5–15% NaCl), and PHA production (2.77–4.54 g L⁻¹). A pullulan synthesized by Aureobasidium pullulans C7 showed a low viscosity rate (from 395 Pa·s to 598 Pa·s) depending on the concentration of polysaccharide solutions. Therefore, at 8.0%, w/v concentration, viscosity virtually remained unchanged with increasing shear rate, indicating that it exhibits Newtonian flow behavior. The effectiveness of various antifungal seed coating formulations has been demonstrated to enhance the tolerance of barley plants to phytopathogens.

Orally delivered probiotics must survive transit through harsh environments during gastrointestinal (GI) digestion and be delivered and released into the target site. The aim of this work was to evaluate the survivability and delivery of gel-encapsulated Lactobacillus rhamnosus GG (LGG) to the colon. New hybrid symbiotic beads alginate/prebiotic pullulan/probiotic LGG were obtained by the extrusion method. The average size of the developed beads was 3401 µm (wet), 921 µm (dry) and the bacterial titer was 10⁹ CFU/g. The morphology of the beads was studied by a scanning electron microscope, demonstrating the structure of the bacterial cellulose shell and loading with probiotics. For the first time, we propose adding an enzymatic extract of feces to an artificial colon fluid, which mimics the total hydrolytic activity of the intestinal microbiota. The beads can be digested by fecalase with cellulase activity, indicating intestinal release. The encapsulation of LGG significantly enhanced their viability under simulated GI conditions. However, the beads, in combination with the prebiotic, provided greater protection of bacteria, enhancing their survival and even increasing cell numbers in the capsules. These data suggest the promising prospects of coencapsulation as an innovative delivery method based on the inclusion of probiotic bacteria in a symbiotic matrix.

This study describes the development of a new combined polysaccharide-matrix-based technology for the immobilization of Lactobacillus rhamnosus GG (LGG) bacteria in biofilm form. The new composition allows for delivering the bacteria to the digestive tract in a manner that improves their robustness compared with planktonic cells and released biofilm cells. Granules consisting of a polysaccharide matrix with probiotic biofilms (PMPB) with high cell density (>9 log CFU/g) were obtained by immobilization in the optimized nutrient medium. Successful probiotic loading was confirmed by fluorescence microscopy and scanning electron microscopy. The developed prebiotic polysaccharide matrix significantly enhanced LGG viability under acidic (pH 2.0) and bile salt (0.3%) stress conditions. Enzymatic extract of feces, mimicking colon fluid in terms of cellulase activity, was used to evaluate the intestinal release of probiotics. PMPB granules showed the ability to gradually release a large number of viable LGG cells in the model colon fluid. In vivo, the oral administration of PMPB granules in rats resulted in the successful release of probiotics in the colon environment. The biofilm-forming incubation method of immobilization on a complex polysaccharide matrix tested in this study has shown high efficacy and promising potential for the development of innovative biotechnologies.

Soil salinity has become a global issue that is directly related to land degradation and results in many changes in climate, ecosystem services, and biodiversity. The present study focuses on the investigation of beneficial properties of a plant probiotic bacterial strain as an eco-friendly and sustainable approach to promote crop growth in saline soil. The endophytic halotolerant strain Pseudomonas flavescens D5 isolated from common chicory (Cichorium intybus L.) was able to grow on a medium containing 15% NaCl; produced indole-3-acetic acid (45.2 μg mL⁻¹) and polyhydroxyalkanoate (1.72 g L⁻¹); and had amylolytic, cellulolytic, and proteolytic activities. Polyhydroxyalkanoate had a pronounced antifungal activity against Fusarium graminearum, F. solani, F. oxysporum, and Alternaria alternata. Under salt stress conditions, inoculation with Ps. flavescens D5 increased the shoot biomass of barley plants by 8–30%, root biomass by 7–20%, chlorophyll a by 18–52%, and chlorophyll b by 7–15%. The content of proline decreased by 1.5–1.8 times. An increase in the activity of antioxidant enzymes (catalase, guaiacol peroxidase, and ascorbate peroxidase) was determined. In inoculated plants growing in saline soil, the content of Na⁺ ions was lower by up to 54.8% compared to control. This strain is promising for stimulating plant growth and protecting them from diseases and other adverse environmental factors, including salt stress.