Properties of Degradable Polyhydroxyalkanoates Synthesized from New Waste Fish Oils (WFOs).

The synthesis of PHA was first investigated using WFOs obtained from smoked-sprat heads, substandard fresh sprats, and fresh mackerel heads and backbones. All the WFOs ensured the growth of the wild-type strain Cupriavidus necator B-10646 and the synthesis of PHA, regardless of the degree of lipid saturation (from 0.52 to 0.65) and the set and ratio of fatty acids (FA), which was represented by acids with chain lengths from C14 to C24. The bacterial biomass concentration and PHA synthesis were comparable (4.1-4.6 g/L and about 70%) when using WFO obtained from smoked-sprat heads and fresh mackerel, and it was twice as high as the bacterial biomass concentration from the fresh sprat waste. This depended on the type of WFO, the bacteria synthesized P(3HB) homopolymer or P(3HB-co-3HV-co-3HHx) copolymer, which had a lower degree of crystallinity (Cx 71%) and a lower molecular weight (Mn 134 kDa) compared to the P(3HB) (Mn 175-209 kDa and Cx 74-78%) at comparable temperatures (Tmelt and Tdegr of 158-168 °C and 261-284 °C, respectively). The new types of WFO, studied for the first time, are suitable as a carbon substrates for PHA synthesis. The WFOs obtained in the production of canned Baltic sprat and Baltic mackerel can be considered a promising and renewable substrate for PHA biosynthesis.

Characteristics of Microparticles Based on Resorbable Polyhydroxyalkanoates Loaded with Antibacterial and Cytostatic Drugs.

The development of controlled drug delivery systems, in the form of microparticles, is an important area of experimental pharmacology. The success of the design and the quality of the obtained microparticles are determined by the method of manufacture and the properties of the material used as a carrier. The goal is to obtain and characterize microparticles depending on their method of preparation, the chemical composition of the polymer and the load of the drugs. To obtain microparticles, four types of degradable PHAs, differing in their chemical compositions, degrees of crystallinity, molecular weights and temperature characteristics, were used (poly-3-hydroxybutyrate and copolymers 3-hydroxybutyric-co-3-hydroxyvaleric acid, 3-hydroxybutyric-co-4-hydroxybutyric acid, and 3-hydroxybutyric-co-3-hydroxyhexanoic acid). The characteristics of microparticles from PHAs were studied. Good-quality particles with an average particle diameter from 0.8 to 65.0 µm, having satisfactory ζ potential values (from -18 to -50 mV), were obtained. The drug loading content, encapsulation efficiency and in vitro release were characterized. Composite microparticles based on PHAs with additives of polyethylene glycol and polylactide-co-glycolide, and loaded with ceftriaxone and 5-fluorouracil, showed antibacterial and antitumor effects in E. coli and HeLa cultures. The results indicate the high potential of PHAs for the design of modern and efficient drug delivery systems.

Biodegradable Polyhydroxyalkanoates with a Different Set of Valerate Monomers: Chemical Structure and Physicochemical Properties.

The properties, features of thermal behavior and crystallization of copolymers containing various types of valerate monomers were studied depending on the set and ratio of monomers. We synthesized and studied the properties of three-component copolymers containing unusual monomers 4-hydroxyvalerate (4HV) and 3-hydroxy-4-methylvalerate (3H4MV), in addition to the usual 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) monomers. The results showed that P(3HB-co-3HV-co-4HV) and P(3HB-co-3HV-co-3H4MV) terpolymers tended to increase thermal stability, especially for methylated samples, including an increase in the gap between melting point (Tmelt) and thermal degradation temperature (Tdegr), an increase in the melting point and glass transition temperature, as well as a lower degree of crystallinity (40-46%) compared with P(3HB-co-3HV) (58-66%). The copolymer crystallization kinetics depended on the set and ratio of monomers. For terpolymers during exothermic crystallization, higher rates of spherulite formation (Gmax) were registered, reaching, depending on the ratio of monomers, 1.6-2.0 µm/min, which was several times higher than the Gmax index (0.52 µm/min) for the P(3HB-co-3HV) copolymer. The revealed differences in the thermal properties and crystallization kinetics of terpolymers indicate that they are promising polymers for processing into high quality products from melts.

Three-Dimensional Printing of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] Biodegradable Scaffolds: Properties, In Vitro and In Vivo Evaluation.

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.

Effect of Monomers of 3-Hydroxyhexanoate on Properties of Copolymers Poly(3-Hydroxybutyrate-co 3-Hydroxyhexanoate).

The properties of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx) copolymers with different ratios of monomers synthesized by the wild-type strain Cupriavidus necator B-10646 on sugars, and an industrial sample from Kaneka synthesized by the recombinant strain C. necator NSDG-ΔfadB1 on soybean oil, were studied in a comparative aspect and in relation to poly(3-hydroxybutyrate) P(3HB). The copolymer samples, regardless of the synthesis conditions or the ratio of monomers, had reduced values of crystallinity degree (50-60%) and weight average molecular weight (415-520 kDa), and increased values of polydispersity (2.8-4.3) compared to P(3HB) (70-76%, 720 kDa, and 2.2). The industrial sample had differences in its thermal behavior, including a lower glass transition temperature (-2.4 °C), two peaks in its crystallization and melting regions, a lower melting point (Tmelt) (112/141 °C), and a more pronounced gap between Tmelt and the temperature of thermal degradation (Tdegr). The process, shape, and size of the spherulites formed during the isothermal crystallization of P(3HB) and P(3HB-co-3HHx) were generally similar, but differed in the maximum growth rate of the spherulites during exothermic crystallization, which was 3.5-3.7 µm/min for P(3HB), and 0.06-1.25 for the P(3HB-co-3HHx) samples. The results from studying the thermal properties and the crystallization mechanism of P(3HB-co-3HHx) copolymers are important for improving the technologies for processing polymer products from melts.

Influence of PHA Substrate Surface Characteristics on the Functional State of Endothelial Cells.

The needs of modern regenerative medicine for biodegradable polymers are wide and varied. Restoration of the viability of the vascular tree is one of the most important components of the preservation of the usefulness of organs and tissues. The creation of vascular implants compatible with blood is an important task of vascular bioengineering. The function of the endothelial layer of the vessel, being largely responsible for the development of thrombotic complications, is of great importance for hemocompatibility. The development of surfaces with specific characteristics of biomaterials that are used in vascular technologies is one of the solutions for their correct endothelialization. Linear polyhydroxyalkanoates (PHAs) are biodegradable structural polymeric materials suitable for obtaining various types of implants and tissue engineering, having a wide range of structural and physicomechanical properties. The use of PHA of various monomeric compositions in endothelial cultivation makes it possible to evaluate the influence of material properties, especially surface characteristics, on the functional state of cells. It has been established that PHA samples with the inclusion of 3-hydroxyhexanoate have optimal characteristics for the formation of a human umbilical vein endothelial cell, HUVEC, monolayer in terms of cell morphology as well as the levels of expression of vinculin and VE-cadherin. The obtained results provide a rationale for the use of PHA copolymers as materials for direct contact with the endothelium in vascular implants.

Modification of Polyhydroxyalkanoates Polymer Films Surface of Various Compositions by Laser Processing.

The results of surface modification of solvent casting films made from polyhydroxyalkanoates (PHAs) of various compositions are presented: homopolymer poly-3-hydroxybutyrate P(3HB) and copolymers comprising various combinations of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 4-hydroxybutyrate(4HB), and 3-hydroxyhexanoate (3HHx) monomers treated with a CO2 laser in continuous and quasi-pulsed radiation modes. The effects of PHAs film surface modification, depending on the composition and ratio of monomers according to the results of the study of SEM and AFM, contact angles of wetting with water, adhesion and growth of fibroblasts have been revealed for the laser radiation regime used. Under continuous irradiation with vector lines, melted regions in the form of grooves are formed on the surface of the films, in which most of the samples have increased values of the contact angle and a decrease in roughness. The quasi-pulse mode by the raster method causes the formation of holes without pronounced melted zones, the total area of which is lower by 20% compared to the area of melted grooves. The number of viable fibroblasts NIH 3T3 on the films after the quasi-pulse mode is 1.5-2.0 times higher compared to the continuous mode, and depends to a greater extent on the laser treatment mode than on the PHAs' composition. The use of various modes of laser modification on the surface of PHAs with different compositions makes it possible to influence the morphology and properties of polymer films in a targeted manner. The results that have been obtained contribute to solving the critical issue of functional biodegradable polymeric materials.

Biosynthesis and Properties of Sulfur-Containing Polyhydroxyalkanoates (PHAs) Produced by Wild-Type Strain Cupriavidus necator B-10646.

The study addresses the growth of the wild-type strain Cupriavidus necator B-10646 and the synthesis of sulfur-containing polyhydroxyalkanoates (PHA) by this strain on media containing fructose and three different precursors (3-mercaptopropionic acid, 3',3'-dithiodipropionic acid and 3',3'-thiodipropionic acid). By varying the concentration and number of doses of the precursors added into the bacterial culture, it was possible to find conditions that ensure the formation of 3-mercaptopropionate (3MP) monomers from the precursors and their incorporation into the C-chain of poly(3-hydroxybutyrate). A series of P(3HB-co-3MP) copolymer samples with different content of 3MP monomers (from 2.04 to 39.0 mol.%) were synthesized and the physicochemical properties were studied. The effect of 3MP monomers is manifested in a certain decrease in the molecular weight of the samples and an increase in polydispersity. Temperature changes are manifested in the appearance of two peaks in the melting region with different intervals regardless of the 3MP content. The studied P(3HB-co-3MP) samples, regardless of the content of 3MP monomers, are characterized by equalization of the ratio of the amorphous and crystalline phases and have a close degree of crystallinity with a minimum of 42%, = and a maximum of 54%.

Resorbable Nanomatrices from Microbial Polyhydroxyalkanoates: Design Strategy and Characterization.

From a series of biodegradable natural polymers of polyhydroxyalkanoates (PHAs)-poly-3-hydroxybutyrate (P(3HB) and copolymers containing, in addition to 3HB monomers, monomers of 3-hydroxyvalerate (3HV), 3-hydroxyhexanoate (3HHx), and 4-hydroxybutyrate (4HB), with different ratios of monomers poured-solvent casting films and nanomembranes with oriented and non-oriented ultrathin fibers were obtained by electrostatic molding. With the use of SEM, AFM, and measurement of contact angles and energy characteristics, the surface properties and mechanical and biological properties of the polymer products were studied depending on the method of production and the composition of PHAs. It has been shown in cultures of mouse fibroblasts of the NIH 3T3 line and diploid human embryonic cells of the M22 line that elastic films and nanomembranes composed of P(3HB-co-4HB) copolymers have high biocompatibility and provide adhesion, proliferation and preservation of the high physiological activity of cells for up to 7 days. Polymer films, namely oriented and non-oriented nanomembranes coated with type 1 collagen, are positively evaluated as experimental wound dressings in experiments on laboratory animals with model and surgical skin lesions. The results of planimetric measurements of the dynamics of wound healing and analysis of histological sections showed the regeneration of model skin defects in groups of animals using experimental wound dressings from P(3HB-co-4HB) of all types, but most actively when using non-oriented nanomembranes obtained by electrospinning. The study highlights the importance of nonwoven nanomembranes obtained by electrospinning from degradable low-crystalline copolymers P(3HB-co-4HB) in the effectiveness of the skin wound healing process.

Biosynthesis and Properties of a P(3HB-co-3HV-co-4HV) Produced by Cupriavidus necator B-10646.

Synthesis of P(3HB-co-3HV-co-4HV) copolymers by the wild-type strain Cupriavidus necator B-10646 on fructose or sodium butyrate as the main C-substrate with the addition of γ-valerolactone as a precursor of 3HV and 4HV monomers was studied. Bacterial cells were cultivated in the modes that enabled production of a series of copolymers with molar fractions of 3HV (from 7.3 to 23.4 mol.%) and 4HV (from 1.9 to 4.7 mol.%) with bacterial biomass concentration (8.2 ± 0.2 g/L) and PHA content (80 ± 2%). Using HPLC, DTA, DSC, X-Ray, SEM, and AFM, the physicochemical properties of copolymers and films prepared from them have been investigated as dependent on proportions of monomers. Copolymers are characterized by a reduced degree of crystallinity (Cx 38-49%) molecular weight characteristics Mn (45-87 kDa), and Mw (201-248 kDa) compared with P(3HB). The properties of the films surface of various composition including the porosity and surface roughness were studied. Most of the samples showed a decrease in the average pore area and an increase in their number with a total increase in 3HV and 4HV monomers. The results allow scaling up the productive synthesis of P(3HB-co-3HV-co-4HV) copolymers using Cupriavidus necator B-10646.

Degradable Poly(3-hydroxybutyrate)-The Basis of Slow-Release Fungicide Formulations for Suppressing Potato Pathogens.

Three-component slow-release fungicide formulations with different modes of action of the active ingredients for suppressing potato pathogens were constructed for the first time. The difenoconazole, mefenoxam, prothioconazole, and azoxystrobin fungicides were embedded in the degradable polymer P(3HB)/birch wood flour blend and examined using SEM, IR spectroscopy, X-ray analysis, DTA, and DSC. Results showed that no chemical bonds were established between the components and that they were physical mixtures that had a lower degree of crystallinity compared to the initial P(3HB), which suggested different crystallization kinetics in the mixtures. The degradation behavior of the experimental formulations was investigated in laboratory micro-ecosystems with pre-characterized field soil. The slow-release fungicide formulations were prolonged-action forms with a half-life of at least 50-60 d, enabling gradual and sustained delivery of the active ingredients to plants. All slow-release fungicide formulations had a strong inhibitory effect on the most common and harmful potato pathogens (Phytophthorainfestans, Alternarialongipes, Rhizoctoniasolani, and Fusariumsolani).

Effectiveness of slow-release fungicide formulations for suppressing potato pathogens.

BACKGROUND: For the first time, the biological activity of slow-release fungicide formulations for suppressing potato pathogens deposited in a degradable poly-3-hydroxybutyrate/sawdust base has been obtained and investigated. RESULTS: The slow-release fungicide formulations (azoxystrobin, azoxystrobin + mefenoxam, and difenoconazole) were studied in vitro and in vivo in comparison with commercial analogues. In in vitro cultures of phytopathogens, the deposited fungicides showed an inhibitory effect comparable to commercial analogues, limiting the growth of colonies of Phytophthora infestans, Alternaria longipes, Rhizoctonia solani and Fusarium solan (2.0-2.3 times relative to the negative control). In laboratory experiments, the use of deposited fungicides was accompanied by earlier germination and more active growth of potatoes against the background of a decrease in the area of plant damage and an increase in yield. In the field experiment, the deposited fungicides suppressed the development of Phytophthora and Alternariosis in the rhizosphere during the entire growing season and reduced the area of plant damage by pathogens by 10-15%, which is two times less than in the groups of plants treated with commercial preparations. The higher biological activity of the embedded fungicides ensured the maximum number of tubers undamaged by pathogens and the total yield of 22-23 t ha-1 , which exceeded the yields in the groups with commercial fungicides (18.4-20.8 t ha-1 ). CONCLUSION: The slow-release fungicide formulations deposited in a degradable P(3HB)/sawdust base are effective in protecting potatoes from pathogens and increasing yields and have an advantage over commercial counterparts. © 2022 Society of Chemical Industry.

Sugar Beet Molasses as a Potential C-Substrate for PHA Production by Cupriavidus necator.

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 ß-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.%.

Bacterial Cellulose (BC) and BC Composites: Production and Properties.

The synthesis of bacterial cellulose (BC) by Komagataeibacter xylinus strain B-12068 was investigated on various C-substrates, under submerged conditions with stirring and in static surface cultures. We implemented the synthesis of BC on glycerol, glucose, beet molasses, sprat oil, and a mixture of glucose with sunflower oil. The most productive process was obtained during the production of inoculum in submerged culture and subsequent growth of large BC films (up to 0.2 m2 and more) in a static surface culture. The highest productivity of the BC synthesis process was obtained with the growth of bacteria on molasses and glycerol, 1.20 and 1.45 g/L per day, respectively. We obtained BC composites with silver nanoparticles (BC/AgNPs) and antibacterial drugs (chlorhexidine, baneocin, cefotaxime, and doripenem), and investigated the structure, physicochemical, and mechanical properties of composites. The disc-diffusion method showed pronounced antibacterial activity of BC composites against E. coli ATCC 25922 and S. aureus ATCC 25923.

Assessment of the efficacy of slow-release formulations of the tribenuron-methyl herbicide in field-grown spring wheat.

The efficacy of slow-release formulations of tribenuron-methyl (TBM) embedded in the matrix of degradable poly(3-hydroxybutyrate) blended with birch wood flour [polymer/wood flour/herbicide 50/30/20 wt.%] was compared with the efficacy of TBM as the active ingredient of the Mortira commercial formulation, which was applied as post-emergence spray to treat spring wheat cv. Novosibirskaya 15. The study was conducted in Central Siberia (in the environs of the city of Krasnoyarsk, Russia) from May to August 2020. The biological efficacy of the embedded TBM was 92.3%, which was considerably higher than the biological efficacy of the Mortira formulation used as the post-emergence spray (15.4%). The embedding of TBM into degradable blended matrix enabled long-duration functioning of this unstable herbicide in soil. The sensitivity of weed plants to TBM differed depending on the species. TBM was more effective against A. retroflexus and A. blitoides, which were killed at an earlier stage, than against C. album and G. aparine, whose percentage increased in the earlier stage and which were controlled by the herbicide less effectively and at later stages. On the plot treated with the embedded herbicide, the parameters of the wheat yield structure were the best, and the total yield was the highest: 3360 ± 40 kg/ha versus 3250 ± 50 kg/ha in the group of plants sprayed with the Mortira formulation. The grain produced in all groups was of high quality and was classified as Grade 1 food grain. The highest quality parameters (grain hectoliter mass, gluten, and protein contents) were obtained in the group of plants treated with the embedded herbicide. The study of the embedded TBM confirmed the high efficacy of the experimental formulation.

Properties of Degradable Polyhydroxyalkanoates (PHAs) Synthesized by a New Strain, Cupriavidus necator IBP/SFU-1, from Various Carbon Sources.

The bacterial strain isolated from soil was identified as Cupriavidus necator IBP/SFU-1 and investigated as a PHA producer. The strain was found to be able to grow and synthesize PHAs under autotrophic conditions and showed a broad organotrophic potential towards different carbon sources: sugars, glycerol, fatty acids, and plant oils. The highest cell concentrations (7-8 g/L) and PHA contents were produced from oleic acid (78%), fructose, glucose, and palm oil (over 80%). The type of the carbon source influenced the PHA chemical composition and properties: when grown on oleic acid, the strain synthesized the P(3HB-co-3HV) copolymer; on plant oils, the P(3HB-co-3HV-co-3HHx) terpolymer, and on the other substrates, the P(3HB) homopolymer. The type of the carbon source influenced molecular-weight properties of PHAs: P(3HB) synthesized under autotrophic growth conditions, from CO2, had the highest number-average (290 ± 15 kDa) and weight-average (850 ± 25 kDa) molecular weights and the lowest polydispersity (2.9 ± 0.2); polymers synthesized from organic carbon sources showed increased polydispersity and reduced molecular weight. The carbon source was not found to affect the degree of crystallinity and thermal properties of the PHAs. The type of the carbon source determined not only PHA composition and molecular weight but also surface microstructure and porosity of the polymer films. The new strain can be recommended as a promising P(3HB) producer from palm oil, oleic acid, and sugars (fructose and glucose) and as a producer of P(3HB-co-3HV) from oleic acid and P(3HB-co-3HV-co-3HHx) from palm oil.

Laser Processing of Polymer Films Fabricated from PHAs Differing in Their Monomer Composition.

The study reports results of using a CO2-laser in continuous wave (3 W; 2 m/s) and quasi-pulsed (13.5 W; 1 m/s) modes to treat films prepared by solvent casting technique from four types of polyhydroxyalkanoates (PHAs), namely poly-3-hydroxybutyrate and three copolymers of 3-hydroxybutyrate: with 4-hydroxybutyrate, 3-hydroxyvalerate, and 3-hydroxyhexanoate (each second monomer constituting about 30 mol.%). The PHAs differed in their thermal and molecular weight properties and degree of crystallinity. Pristine films differed in porosity, hydrophilicity, and roughness parameters. The two modes of laser treatment altered these parameters and biocompatibility in diverse ways. Films of P(3HB) had water contact angle and surface energy of 92° and 30.8 mN/m, respectively, and average roughness of 144 nm. The water contact angle of copolymer films decreased to 80-56° and surface energy and roughness increased to 41-57 mN/m and 172-290 nm, respectively. Treatment in either mode resulted in different modifications of the films, depending on their composition and irradiation mode. Laser-treated P(3HB) films exhibited a decrease in water contact angle, which was more considerable after the treatment in the quasi-pulsed mode. Roughness parameters were changed by the treatment in both modes. Continuous wave line-by-line irradiation caused formation of sintered grooves on the film surface, which exhibited some change in water contact angle (76-80°) and reduced roughness parameters (to 40-45 mN/m) for most films. Treatment in the quasi-pulsed raster mode resulted in the formation of pits with no pronounced sintered regions on the film surface, a more considerably decreased water contact angle (to 67-76°), and increased roughness of most specimens. Colorimetric assay for assessing cell metabolic activity (MTT) in NIH 3T3 mouse fibroblast culture showed that the number of fibroblasts on the films treated in the continuous wave mode was somewhat lower; treatment in quasi-pulsed radiation mode caused an increase in the number of viable cells by a factor of 1.26 to 1.76, depending on PHA composition. This is an important result, offering an opportunity of targeted surface modification of PHA products aimed at preventing or facilitating cell attachment.

Production and Properties of Microbial Polyhydroxyalkanoates Synthesized from Hydrolysates of Jerusalem Artichoke Tubers and Vegetative Biomass.

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 Tmelt and Tdegr, prevailing of the crystalline phase over the amorphous one (Cx 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.

Metabolic activity of cryogenic soils in the subarctic zone of Siberia towards "green" bioplastics.

The present study investigates, for the first time, the structure of the microbial community of cryogenic soils in the subarctic region of Siberia and the ability of the soil microbial community to metabolize degradable microbial bioplastic - poly-3-hydroxybutyrate [P(3HB)]. When the soil thawed, with the soil temperature between 5-7 and 9-11 °C, the total biomass of microorganisms at a 10-20-cm depth was 226-234 mg g-1 soil and CO2 production was 20-46 mg g-1 day-1. The total abundance of microscopic fungi varied between (7.4 ± 2.3) × 103 and (18.3 ± 2.2) × 103 CFU/g soil depending on temperature; the abundance of bacteria was several orders of magnitude greater: (1.6 ± 0.1) × 106 CFU g-1 soil. The microbial community in the biofilm formed on the surface of P(3HB) films differed from the background soil in concentrations and composition of microorganisms. The activity of microorganisms caused changes in the surface microstructure of polymer films, a decrease in molecular weight, and an increase in the degree of crystallinity of P(3HB), indicating polymer biodegradation due to metabolic activity of microorganisms. The clear-zone technique - plating of isolates on the mineral agar with polymer as sole carbon source - was used to identify P(3HB)-degrading microorganisms inhabiting cryogenic soil in Evenkia. Analysis of nucleotide sequences of rRNA genes was performed to identify the following P(3HB)-degrading species: Bacillus pumilus, Paraburkholderia sp., Pseudomonas sp., Rhodococcus sp., Stenotrophomonas rhizophila, Streptomyces prunicolor, and Variovorax paradoxus bacteria and the Penicillium thomii, P. arenicola, P. lanosum, Aspergillus fumigatus, and A. niger fungi.

Constructing sustained-release herbicide formulations based on poly-3-hydroxybutyrate and natural materials as a degradable matrix.

BACKGROUND: The purpose of the present study was to develop ecofriendly herbicide formulations. Its main aim was to develop and investigate slow-release formulations of herbicides (metribuzin, tribenuron-methyl, and fenoxaprop-P-ethyl) of different structure, solubility, and specificity, which were loaded into a degradable matrix of poly-3-hydroxybutyrate (P(3HB)) blended with available natural materials (peat, clay, and wood flour). RESULTS: Differences in the structure and physicochemical properties of the formulations were studied depending on the type of the matrix. Herbicide release and accumulation in soil were associated with the solubility of the herbicide. Fourier-transform infrared spectroscopy showed that no chemical bonds were formed between the components in the experimental formulations. Degradation of the formulations in agro-transformed soil in laboratory conditions was chiefly influenced by the shape of the specimens (granules or pellets) while the effect of the type of filler (peat, clay, or wood flour) was insignificant. The use of granules enabled more rapid accumulation of the herbicides in soil: their peak concentrations were reached after 3 weeks of incubation while the concentrations of the herbicides released from the pellets were the highest after 5-7 weeks. Loading of the herbicides into the polymer matrix composed of the slowly degraded P(3HB) and natural materials enabled both sustained function of the formulations in soil (lasting between 1.5 and ≥3 months) and stable activity of the otherwise rapidly inactivated herbicides such as tribenuron-methyl and fenoxaprop-P-ethyl. CONCLUSION: The experimental herbicide formulations enabled slow release of the active ingredients to soil. © 2019 Society of Chemical Industry.