To increase the availability and expand the raw material base, the production of polyhydroxyalkanoates (PHA) by the wild strain Cupriavidus necator B-10646 on hydrolysates of sugar beet molasses was studied. The hydrolysis of molasses was carried out using β-fructofuranosidase, which provides a high conversion of sucrose (88.9%) to hexoses. We showed the necessity to adjust the chemical composition of molasses hydrolysate to balance with the physiological needs of C. necator B-10646 and reduce excess sugars and nitrogen and eliminate phosphorus deficiency. The modes of cultivation of bacteria on diluted hydrolyzed molasses with the controlled feeding of phosphorus and glucose were implemented. Depending on the ratio of sugars introduced into the bacterial culture due to the molasses hydrolysate and glucose additions, the bacterial biomass concentration was obtained from 20–25 to 80–85 g/L with a polymer content up to 80%. The hydrolysates of molasses containing trace amounts of propionate and valerate were used to synthesize a P(3HB-co-3HV) copolymer with minor inclusions of 3-hydroxyvlaerate monomers. The introduction of precursors into the medium ensured the synthesis of copolymers with reduced values of the degree of crystallinity, containing, in addition to 3HB, monomers 3HB, 4HB, or 3HHx in an amount of 12–16 mol.%.

One of the major challenges in PHA biotechnology is optimization of biotechnological processes of the entire synthesis, mainly by using new inexpensive carbon substrates. A promising substrate for PHA synthesis may be the sugars extracted from the Jerusalem artichoke. In the present study, hydrolysates of Jerusalem artichoke (JA) tubers and vegetative biomass were produced and used as carbon substrate for PHA synthesis. The hydrolysis procedure (the combination of aqueous extraction and acid hydrolysis, process temperature and duration) influenced the content of reducing substances (RS), monosaccharide contents, and the fructose/glucose ratio. All types of hydrolysates tested as substrates for cultivation of three strains—C. necator B-10646 and R. eutropha B 5786 and B 8562—were suitable for PHA synthesis, producing different biomass concentrations and polymer contents. The most productive process, conducted in 12-L fermenters, was achieved on hydrolysates of JA tubers (X = 66.9 g/L, 82% PHA) and vegetative biomass (55.1 g/L and 62% PHA) produced by aqueous extraction of sugars at 80 °C followed by acid hydrolysis at 60 °C, using the most productive strain, C. necator B-10646. The effects of JA hydrolysates on physicochemical properties of PHAs were studied for the first time. P(3HB) specimens synthesized from the JA hydrolysates, regardless of the source (tubers or vegetative biomass), hydrolysis conditions, and PHA producing strain employed, exhibited the 100–120 °C difference between the T_melt and T_degr, prevailing of the crystalline phase over the amorphous one (C_x between 69 and 75%), and variations in weight average molecular weight (409–480) kDa. Supplementation of the culture medium of C. necator B-10646 grown on JA hydrolysates with potassium valerate and ε-caprolactone resulted in the synthesis of P(3HB-co-3HV) and P(3HB-co-4HB) copolymers that had decreased degrees of crystallinity and molecular weights, which influenced the porosity and surface roughness of polymer films prepared from them. The study shows that JA hydrolysates used as carbon source enabled productive synthesis of PHAs, comparable to synthesis from pure sugars. The next step is to scale up PHA synthesis from JA hydrolysates and conduct the feasibility study. The present study contributes to the solution of the critical problem of PHA biotechnology—finding widely available and inexpensive substrates.

The results of constructing 3D scaffolds from degradable poly(3-hydrosbutyrpate-co-3-hydroxyvalerate) using FDM technology and studying the structure, mechanical properties, biocompatibility in vitro, and osteoplastic properties in vivo are presented. In the process of obtaining granules, filaments, and scaffolds from the initial polymer material, a slight change in the crystallization and glass transition temperature and a noticeable decrease in molecular weight (by 40%) were registered. During the compression test, depending on the direction of load application (parallel or perpendicular to the layers of the scaffold), the 3D scaffolds had a Young’s modulus of 207.52 ± 19.12 and 241.34 ± 7.62 MPa and compressive stress tensile strength of 19.45 ± 2.10 and 22.43 ± 1.89 MPa, respectively. SEM, fluorescent staining with DAPI, and calorimetric MTT tests showed the high biological compatibility of scaffolds and active colonization by NIH 3T3 fibroblasts, which retained their metabolic activity for a long time (up to 10 days). The osteoplastic properties of the 3D scaffolds were studied in the segmental osteotomy test on a model defect in the diaphyseal zone of the femur in domestic Landrace pigs. X-ray and histological analysis confirmed the formation of fully mature bone tissue and complete restoration of the defect in 150 days of observation. The results allow us to conclude that the constructed resorbable 3D scaffolds are promising for bone grafting.

Biodegradable polyhydroxyalkanoates, biopolymers of microbiological origin, formed by 3- and 4-hydroxybutyrate monomers P(3HB-co-4HB), were used to obtain nanomembranes loaded with drugs as cell carriers by electrospinning. Resorbable non-woven membranes from P(3HB-co-4HB) loaded with ceftazidime, doripinem, and actovegin have been obtained. The loading of membranes with drugs differently affected the size of fibers and the structure of membranes, and in all cases increased the hydrophilicity of the surface. The release of drugs in vitro was gradual, which corresponded to the Higuchi and Korsmeyer-Peppas models. Antibiotic-loaded membranes showed antibacterial activity against S. aureus and E. coli, in which growth inhibition zones were 41.7 ± 1.1 and 38.6 ± 1.7 mm for ceftazidime and doripinem, respectively. The study of the biological activity of membranes in the NIH 3T3 mouse fibroblast culture based on the results of DAPI and FITC staining of cells, as well as the MTT test, did not reveal a negative effect despite the presence of antibiotics in them. Samples containing actovegin exhibit a stimulating effect on fibroblasts. Biodegradable polyhydroxyalkanoates formed by 3-hydroxybutyrate and 4-hydroxybutyrate monomers provide electrospinning non-woven membranes suitable for long-term delivery of drugs and cultivation of eukaryotic cells, and are promising for the treatment of wound defects complicated by infection.

The article presents the results of a study of constructed composites based on degradable poly(3-hydroxybutyrate) (P(3HB)) filled with plant materials of 30, 50, and 70% of different origin—wood flour (WF) from birch (Betula pendula), hemp hurds (HH) or hemp fiber (HF) (Cannabis sativa). Composite bar samples were obtained by hot pressing homogeneous mixtures of polymer and fillers at 170 °C and a specific pressure of 6.13 MPa. The influence of the filler type and the polymer/filler ratio on the temperature characteristics of the samples, density, microstructure, surface properties, water absorption, physical and mechanical properties, and degradability in soil was determined. The Young’s modulus of the samples ranged from 2640 to 3715 MPa, depending on the composition. The maximum degradation of the composites after 120 days of exposure to soil was recorded at 70% WF, HH, or HF filling, amounting to 77.4, 63.5, and 38.6%, respectively. Perspective biodegradable composites based on P(3HB) filled with various plant-based fillers were obtained and characterized, along with new knowledge about their properties, the lack of which currently hinders the active development and commercialization of such in-demand materials.