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.

Fermentative bioconversion of food waste into biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) using Cupriavidus necator.

Dependency on plastic commodities has led to a recurrent increase in their global production every year. Conventionally, plastic products are derived from fossil fuels, leading to severe environmental concerns. The recent coronavirus disease 2019 pandemic has triggered an increase in medical waste. Conversely, it has disrupted the supply chain of personal protective equipment (PPE). Valorisation of food waste was performed to cultivate C. necator for fermentative production of biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). The increase in biomass, PHBV yield and molar 3-hydroxy valerate (3HV) content was estimated after feeding volatile fatty acids. The fed-batch fermentation strategy reported in this study produced 15.65 ± 0.14 g/L of biomass with 5.32 g/L of PHBV with 50% molar 3HV content. This is a crucial finding, as molar concentration of 3HV can be modulated to suit the specification of biopolymer (film or fabric). The strategy applied in this study addresses the issue of global food waste burden and subsequently generates biopolymer PHBV, turning waste to wealth.

Interphase Design of Cellulose Nanocrystals/Poly(hydroxybutyrate-ran-valerate) Bionanocomposites for Mechanical and Thermal Properties Tuning.

Poly[(3-hydroxybutyrate)-ran-(3-hydroxyvalerate)] (PHBV) is a bacterial polyester with a strong potential as a substitute for oil-based thermoplastics due to its biodegradability and renewability. However, its inherent slow crystallization rate limits its thermomechanical properties and therefore its applications. In this work, surface-modified cellulose nanocrystals (CNCs) have been investigated as green and biosourced nucleating and reinforcing agent for PHBV matrix. Different ester moieties from the CNCs were thereby produced through a green one-pot hydrolysis/Fisher esterification. Beyond the improved dispersion, the CNCs surface esterification affected the thermal and thermomechanical properties of PHBV. The results demonstrate that butyrate-modified CNCs, mimicking the PHBV chemical structure, brought a considerable improvement toward the CNCs/matrix interface, leading to an enhancement of the PHBV thermomechanical properties via a more efficient stress transfer, especially above its glass transition.

Efficient production and characterization of homopolymeric poly(3-hydroxyvalerate) produced by Bacillus strain PJC48.

Aliphatic polyester, poly(3-hydroxyvalerate) (PHV), is commonly produced as a granular component in bacterial cells of various species. Based on 16S rDNA gene sequence analysis, strain PJC48 was identified as a Bacillus species. The current study is aimed to screen for a high-yield strain that can produce PHV efficiently and to increase PHV product yield by optimizing the fermentative process. We identified a high-producer strain based on Nile red staining. Characterization of the PHV produced by PJC48 by nuclear magnetic resonance spectroscopy revealed that it consisted of (R)-3-hydroxyvalerate monomers. The suggested model was validated by response surface methodology. Optimization of the PHV yield resulted in an increase of 32.75% compared to control, with a maximum production of 1.64 g/L after 48 H.

Salivary Hydrogen Sulfide Measured with a New Highly Sensitive Self-Immolative Coumarin-Based Fluorescent Probe.

Ample evidence suggests that H2S is an important biological mediator, produced by endogenous enzymes and microbiota. So far, several techniques including colorimetric methods, electrochemical analysis and sulfide precipitation have been developed for H2S detection. These methods provide sensitive detection, however, they are destructive for tissues and require tedious sequences of preparation steps for the analyzed samples. Here, we report synthesis of a new fluorescent probe for H2S detection, 4-methyl-2-oxo-2H-chromen-7-yl 5-azidopentanoate (1). The design of 1 is based on combination of two strategies for H2S detection, i.e., reduction of an azido group to an amine in the presence of H2S and intramolecular lactamization. Finally, we measured salivary H2S concentration in healthy, 18⁻40-year-old volunteers immediately after obtaining specimens. The newly developed self-immolative coumarin-based fluorescence probe (C15H15N3O4) showed high sensitivity to H2S detection in both sodium phosphate buffer at physiological pH and in saliva. Salivary H2S concentration in healthy volunteers was within a range of 1.641⁻7.124 µM.

Effects of matrix types on formation and transformation of energy-accumulating substances in enhanced biological phosphorus removal (EBPR).

Enhanced biological phosphorus removal (EBPR) has been widely used in wastewater treatment. In this study, a laboratory investigation of activated sludge in A/O-SBR reactor was conducted to probe the effects of the matrix types on EBPR polyphosphate, intracellular polysaccharide, polyhydroxyalkanoates (PHA) formation and transformation. There is a decrease in anaerobic condition and an increase in aerobic condition for the intracellular glycogen of sodium propionate matrix and sodium acetate matrix. While the intracellular glycogen of glucose matrix shows a decreasing tendency in both anaerobic and aerobic reaction process. Sodium acetate matrix is beneficial to the formation of polyhydroxybutyrate (PHB), but the content of PHB is relatively small. PHB and poly-3-hydroxyvalerate (PHV) contents in PHA are quite similar in both anaerobic and aerobic reactions with a PHB/PHV ratio of 0.83-1.45. The synthesis of PHV and PHB is mainly in the initial anaerobic stage (0 h - 1 h). Glucose matrix is helpful to the formation of PHV. The content of polymphosphorus shows an increasing tendency in both anaerobic and aerobic stages, suggesting that glucose matrix acclimation of the reactor favors the formation of polymphosphorus.

Effect of ß-hydroxy-ß-methylbutyrate in masticatory muscles of rats.

The aim of this research was to examine the influence of ß-hydroxy-ß-methylbutyrate (HMB) on changes in the profile of muscle fibers, whether these alterations were similar between the elevator and depressor muscles of the jaw, and whether the effects would be similar in male and female animals. Fifty-eight rats aged 60 days (29 animals of each gender) were divided into four groups: the initial control group (ICG) was sacrificed at the beginning of the experiment; the placebo control group (PCG) received saline and was fed ad libitum; the experimental group (EG) received 0.3 g kg(-1) of HMB daily for 4 weeks by gavage as well as the same amount of food consumed by the PCG in the previous day; and the experimental ad libitum group (EAG) received the same dose of the supplement along with food ad libitum. Samples included the digastric and masseter muscles for the histoenzymological analysis. Data were subjected to statistical analysis with a significance level of P < 0.05. Use of HMB caused a decrease in the percentage of fast twitch glycolytic (FG) fibers and an increase in fast twitch oxidative glycolytic (FOG) fibers in males in both experimental groups (EG and EAG). However, it produced no increase in the muscle fiber area, in either gender, in the masseter muscle. In the digastric muscle, the HMB did not change the frequency or the area of any muscle fiber types in either gender. Our data suggest that the use of HMB caused small changes in the enzymological profile of fibers of the mastication muscles; the changes were different in the elevator and depressor muscles of the jaw and the results were different depending on gender.

3-Hydroxybutyrate oligomer hydrolase and 3-hydroxybutyrate dehydrogenase participate in intracellular polyhydroxybutyrate and polyhydroxyvalerate degradation in Paracoccus denitrificans.

Genes encoding 3-hydroxybutyrate oligomer hydrolase (PhaZc) and 3-hydroxybutyrate dehydrogenase (Hbd) were isolated from Paracoccus denitrificans. PhaZc and Hbd were overproduced as His-tagged proteins in Escherichia coli and purified by affinity and gel filtration chromatography. Purified His-tagged proteins had molecular masses of 31 kDa and 120 kDa (a tetramer of 29-kDa subunits). The His-tagged PhaZc hydrolyzed not only 3-hydroxybutyrate oligomers but also 3-hydroxyvalerate oligomers. The His-tagged Hbd catalyzed the dehydrogenation of 3-hydroxyvalerate as well as 3-hydroxybutyrate. When both enzymes were included in the same enzymatic reaction system with 3-hydroxyvalerate dimer, sequential reactions occurred, suggesting that PhaZc and Hbd play an important role in the intracellular degradation of poly(3-hydroxyvalerate). When the phaZc gene was disrupted in P. denitrificans by insertional inactivation, the mutant strain lost PhaZc activity. When the phaZc-disrupted P. denitrificans was complemented with phaZc, PhaZc activity was restored. These results suggest that P. denitrificans carries a single phaZc gene. Disruption of the phaZc gene in P. denitrificans affected the degradation rate of PHA.

Biosynthesis of polyhydroxyalkanoate copolymers from methanol by Methylobacterium extorquens AM1 and the engineered strains under cobalt-deficient conditions.

Methylobacterium extorquens AM1 has been shown to accumulate polyhydroxyalkanoate (PHA) composed solely of (R)-3-hydroxybutyrate (3HB) during methylotrophic growth. The present study demonstrated that the wild-type strain AM1 grown under Co²âº-deficient conditions accumulated copolyesters of 3HB and a C5-monomer, (R)-3-hydroxyvalerate (3HV), using methanol as the sole carbon source. The 3HV unit was supposed to be derived from propionyl-CoA, synthesized via the ethylmalonyl-CoA pathway impaired by Co²âº limitation. This assumption was strongly supported by the dominant incorporation of the 3HV unit into PHA when a strain lacking propionyl-CoA carboxylase was incubated with methanol. Further genetic engineering of M. extorquens AM1 was employed for the methylotrophic synthesis of PHA copolymers. A recombinant strain of M. extorquens AM1C(Ac) in which the original PHA synthase gene phaC(Me) had been replaced by phaC(Ac), encoding an enzyme with broad substrate specificity from Aeromonas caviae, produced a PHA terpolymer composed of 3HB, 3HV, and a C6-monomer, (R)-3-hydroxyhexanoate, from methanol. The cellular content and molecular weight of the PHA accumulated in the strain AM1C(Ac) were higher than those of PHA in the wild-type strain. The triple deletion of three PHA depolymerase genes in M. extorquens AM1C(Ac) showed no significant effects on growth and PHA biosynthesis properties. Overexpression of the genes encoding ß-ketothiolase and NADPH-acetoacetyl-CoA reductase increased the cellular PHA content and 3HV composition in PHA, although the cell growth on methanol was decreased. This study opens up the possibility of producing practical PHA copolymers with methylotrophic bacteria using methanol as a feedstock.

Cell attachment on poly(3-hydroxybutyrate)-poly(ethylene glycol) copolymer produced by Azotobacter chroococcum 7B.

BACKGROUND: The improvement of biomedical properties, e.g. biocompatibility, of poly(3-hydroxyalkanoates) (PHAs) by copolymerization is a promising trend in bioengineering. We used strain Azotobacter chroococcum 7B, an effective producer of PHAs, for biosynthesis of not only poly(3-hydroxybutyrate) (PHB) and its main copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-HV), but also alternative copolymer, poly(3-hydroxybutyrate)-poly(ethylene glycol) (PHB-PEG). RESULTS: In biosynthesis we used sucrose as the primary carbon source and valeric acid or poly(ethylene glycol) 300 (PEG 300) as additional carbon sources. The chemical structure of PHB-PEG and PHB-HV was confirmed by 1H nuclear-magnetic resonance (1H NMR) analysis. The physico-chemical properties (molecular weight, crystallinity, hydrophilicity, surface energy) and surface morphology of films from PHB copolymers were studied. To study copolymers biocompatibility in vitro the protein adsorption and COS-1 fibroblasts growth on biopolymer films by XTT assay were analyzed. Both copolymers had changed physico-chemical properties compared to PHB homopolymer: PHB-HV and PHB-PEG had less crystallinity than PHB; PHB-HV was more hydrophobic than PHB in contrast to PHB-PEG appeared to have greater hydrophilicity than PHB; whereas the morphology of polymer films did not differ significantly. The protein adsorption to PHB-PEG was greater and more uniform than to PHB and PHB-PEG copolymer promoted better growth of COS-1 fibroblasts compared with PHB homopolymer. CONCLUSIONS: Thus, despite low EG-monomers content in bacterial origin PHB-PEG copolymer, this polymer demonstrated significant improvement in biocompatibility in contrast to PHB and PHB-HV copolymers, which may be coupled with increased protein adsorption and hydrophilicity of PEG-containing copolymer.

Biosynthesis and characterization of a new copolymer, poly(3-hydroxyvalerate-co-5-hydroxydecenoate), from Pseudomonas aeruginosa.

Using a new isolate of Pseudomonas aeruginosa, we obtained 7 g cell dry wt (CDW/l) using 5 % (w/v) glucose. Crude polyhydroxyalkanoates were obtained at 14.6 % of CDW. FTIR and NMR analysis confirmed that this was a new co-polymer: 3-hydroxyvalerate-co-5-hydroxydecenoate. Differential scanning calorimetry analysis showed two different melting temperatures of the copolymer and also indicated the glass transition temperature to be 4 °C. The polydispersity index of the polymer was 1.059.

Production and characterization of a biodegradable polymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), using the type II methanotroph, Methylocystis sp. MJC1.

The production of polyhydroxyalkanoates (PHAs) through the biological conversion of methane is a promising solution to address both methane emissions and plastic waste. Type II methanotrophs naturally accumulate a representative PHA, poly(3-hydroxybutyrate) (PHB), using methane as the sole carbon source. In this study, we aimed to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV copolymer) with improved properties compared with PHB, using the type II methanotroph, Methylocystis sp. MJC1. We optimized the pH, valerate concentration, and valerate supply time in a one-step cultivation process using a gas bioreactor to enhance PHBV copolymer production yield and the 3-hydroxyvalerate (3HV) molar fraction. Under the optimal conditions, the biomass reached 21.3 g DCW/L, and PHBV copolymer accumulation accounted for 41.9 % of the dried cell weight, with a 3HV molar fraction of 28.4 %. The physicochemical properties of the purified PHBV copolymer were characterized using NMR, FTIR, TGA, DSC, and GPC.

Genetically modified strains of Ralstonia eutropha H16 with ß-ketothiolase gene deletions for production of copolyesters with defined 3-hydroxyvaleric acid contents.

ß-Ketothiolases catalyze the first step of poly(3-hydroxybutyrate) [poly(3HB)] biosynthesis in bacteria by condensation of two acetyl coenzyme A (acetyl-CoA) molecules to acetoacetyl-CoA and also take part in the degradation of fatty acids. During growth on propionate or valerate, Ralstonia eutropha H16 produces the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [poly(3HB-co-3HV)]. In R. eutropha, 15 ß-ketothiolase homologues exist. The synthesis of 3-hydroxybutyryl-CoA (3HB-CoA) could be significantly reduced in an 8-fold mutant (Lindenkamp et al., Appl. Environ. Microbiol. 76:5373-5382, 2010). In this study, a 9-fold mutant deficient in nine ß-ketothiolase gene homologues (phaA, bktB, H16_A1713, H16_B1771, H16_A1528, H16_B0381, H16_B1369, H16_A0170, and pcaF) was generated. In order to examine the polyhydroxyalkanoate production capacity when short- or long-chain and even- or odd-chain-length fatty acids were provided as carbon sources, the growth and storage behavior of several mutants from the previous study and the newly generated 9-fold mutant were analyzed. Propionate, valerate, octanoate, undecanoic acid, or oleate was chosen as the sole carbon source. On octanoate, no significant differences in growth or storage behavior were observed between wild-type R. eutropha and the mutants. In contrast, during the growth on oleate of a multiple mutant lacking phaA, bktB, and H16_A0170, diminished poly(3HB) accumulation occurred. Surprisingly, the amount of accumulated poly(3HB) in the multiple mutants grown on gluconate differed; it was much lower than that on oleate. The ß-ketothiolase activity toward acetoacetyl-CoA in H16ΔphaA and all the multiple mutants remained 10-fold lower than the activity of the wild type, regardless of which carbon source, oleate or gluconate, was employed. During growth on valerate as a sole carbon source, the 9-fold mutant accumulated almost a poly(3-hydroxyvalerate) [poly(3HV)] homopolyester with 99 mol% 3HV constituents.

Mechanical recyclability of biodegradable polymers used for food packaging: case study of polyhydroxybutyrate-co-valerate (PHBV) plastic.

For the purpose of mechanical recycling for food contact applications, decontamination of polyhydroxybutyrate-co-valerate (PHBV) plastic was performed under different temperatures and time conditions. As expected, increasing the decontamination temperature and duration increased the decontamination efficiency, but also the degradation of the polymer. The combination 160 °C/6 h was selected as the optimal conditions that maximize contaminants removal while minimizing polymer degradation. Then the safety of the recycled PHBV under these conditions was assessed, in accordance with EFSA regulation based on bottle-to-bottle PET recycling. Decontamination of low molecular weight molecules such as toluene, chlorobenzene, and methyl salicylate was nearly complete with residual concentrations below the modeled concentrations allowed in the polymer when the adult scenario is considered. However, the higher molecular weight and lower volatility molecules exhibited acceptable decontamination efficiencies, but their residual concentrations in the polymer exceeded the maximum concentrations of no concern. The presence of these molecules allows the use of nearly 21% recycled PHBV in the new materials to meet safety criteria. It is important to keep in mind that this work, never done before, is a preliminary work on mechanical recycling of PHBV, mainly based on extrapolation of PET conditions and regulations. Much more research needs to be done to improve the decontamination process, the barrier properties of PHBV or to think about a short recycling line for PHBV.

Production of polyhydroxyalkanoates by three novel species of Marinobacterium.

Several species of novel marine bacteria from the genus Marinobacterium, including M. nitratireducens, M. sediminicola, and M. zhoushanense were found to be capable of producing polyhydroxyalkanoates (PHA) using sugars and volatile fatty acids (VFAs) as the carbon source. M. zhoushanense produced poly-3-hydroxybutytate (PHB) from sucrose, achieving a product titer and PHB content of 2.89 g/L and 64.05 wt%, respectively. By contrast, M. nitratireducens accumulated 3.38 g/L PHB and 66.80 wt% polymer content using butyrate as the substrate. A third species, M. sediminicola showed favorable tolerance to propionate, butyrate, and valerate. The use of 10 g/L valerate yielded 3.37 g/L poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), with a 3-hydroxyvalerate (3 HV) monomer content of 94.75 mol%. Moreover, M. sediminicola could be manipulated to produce PHBV with changeable polymer compositions by feeding different mixtures of VFAs. Our results indicate that M. sediminicola is a promising halophilic bacterium for the production of PHA.

Engineering alginate hydrogel films with poly(3-hydroxybutyrate-co-3-valerate) and graphene nanoplatelets: Enhancement of antiviral activity, cell adhesion and electroactive properties.

A new biodegradable semi-interpenetrated polymer network (semi-IPN) of two US Food and Drug Administration approved materials, poly(3-hydroxybutyrate-co-3-valerate) (PHBV) and calcium alginate (CA) was engineered to provide an alternative strategy to enhance the poor adhesion properties of CA. The synthesis procedure allows the additional incorporation of 10 % w/w of graphene nanoplatelets (GNPs), which have no cytotoxic effect on human keratinocytes. This quantity of multilayer graphene provides superior antiviral activity to the novel semi-IPN against a surrogate virus of SARS-CoV-2. Adding GNPs hardly affects the water absorption or electrical conductivity of the pure components of CA and PHBV. However, the semi-IPN's electrical conductivity increases dramatically after adding GNP due to molecular rearrangements of the intertwined polymer chains that continuously distribute the GNP nanosheets, This new hydrophilic composite biomaterial film shows great promise for skin biomedical applications, especially those that require antiviral and/or biodegradable electroconductive materials.

Synthesis and characterization of PHV-block-mPEG diblock copolymer and its formation of amphiphilic nanoparticles for drug delivery.

Despite the recent research interest in the field of nanoparticles delivery system, their structure modification and transport behavior of various hydrophobic drugs is poorly developed. In this article the synthesis of novel amphiphilic diblock copolymer poly([R]-3-hydroxyvalerate)-block-monomethoxy poly(ethylene glycol) (PHV-block-mPEG) was undertaken by modifying the structure of biodegradable and hydrophobic poly([R]-3-hydroxyvalerate) (PHV) with hydrophilic monomethoxy poly(ethylene glycol) (mPEG). The chemical combination of the two blocks was carried out in the melt using bis(2-ethylhexanoate) tin as transesterification catalyst. The synthesized product was characterized by gel permeation chromatography (GPC), 1H nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC) analysis. The block copolymer self-assembled into amphiphilic nanoparticles with a core of hydrophobic PHV and a shell of hydrophilic mPEG in aqueous solution. Characterization of the nanoparticles showed the formation of discrete, spherically shaped nanoparticles with mean particle size of 200 +/- 1 nm and zeta potential of -14 +/- 1 mV. A hydrophobic drug thymoquinone was efficiently incorporated into the core hydrophobic domain of the nanoparticles and its release kinetics was studied in vitro. The amphiphilic PEGylated nanoparticles showed biocompatibility when checked in neuronal hippocampal cells of prenatal rat. Our results suggest that the amphiphilic nanoparticles with core-shell structures are potentially useful to develop novel drug carriers.

[Composite biodegradable materials based on polyhydroxyalkanoate].

Conditions for the processing and mixing of biodegradable polymers at temperatures less than their thermal destruction (130-150 degrees C) using standard equipment have been identified. The structure of the polyhydroxybutyrate/valerate (PHB/V) copolymer has been revealed and peculiarities of the crystal phase formation at different monomer ratios have been investigated. It was shown that pure PHB with molecular mass 180-270 kDa has elastic module approximately 1.2 GPa, strength approximately 25 MPa, and elongation at break approximately 10%. The most active biodestructors of PHB, PHB/V, and their composites have been selected (Aspergillus caespitosus), and the ability of basidiomycete Panus tigrinus to biodegrade polyalkanoates was demonstrated for the first time. It was shown that A. caespitosus degraded PHB/V and Biopol films along with the PHB with the destruction rate depending on the technology of the film production, on the molecular mass, and on the extend of the polymer crystallinity.

Structure and properties of lipase activated by cellulose-silica polyethersulfone membrane for production of pentyl valerate.

Herein, this study extracted nanocrystalline cellulose (NC) and silica (SiO2) from raw oil palm leaves (OPL), and employed as nanofillers in polyethersulfone (PES) to produce NC-SiO2-PES as support to immobilize Candida rugosa lipase (CRL) (NC-SiO2-PES/CRL). XRD, TGA-DTG and FTIR-ATR data affirmed that NC and SiO2 were isolated from OPL with corresponding crystallinity indices of 68 % and 70 %. A 0.02 cm membrane size with 5% (w/v) of NC-SiO2 without PVP K30 was optimal for membrane fabrication. CRL immobilized on the Glut-AP-NC-SiO2-PES membrane gave a higher conversion of pentyl valerate (PeVa) (91.3 %, p < 0.05) compared to Glut-NC-SiO2-PES (73.9 %) (p < 0.05). Characterization of the NC-SiO2-PES/CRL biocatalyst verified the presence of CRL. Hence, raw OPL is a proven good source of NC and SiO2, as reinforcement nanofillers in PES. The overall findings envisage the promising use of NC-SiO2-PES/CRL to catalyze an expedient and high yield of PeVa, alongside the suitability of NC-SiO2-PES for activating other enzymes.

Biosynthesis and characterization of 3-hydroxyalkanoate terpolyesters with adjustable properties by Aeromonas hydrophila.

Polyhydroxyalkanoates (PHA) terployesters P(3HB-co-3HV-co-3HHx) consisting of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyhexanoate (3HHx) were produced by wild-type Aeromonas hydrophila 4AK4, its recombinant harboring PHA synthesis genes phaPCJ encoding PHA binding protein phasin, PHA synthase, and enoyl-CoA hydratase, and another its recombinant harboring phaAB encoding beta-ketothiolase and acetoacetyl-CoA reductase, respectively, when grown in lauric acid and/or valerate. The terpolyesters produced by A. hydrophila 4AK4 (phaAB) grown in velarate were found to produce copolymers P(3HB-co-3HV) containing high 3HV fractions with a maximum of 99 mol% 3HV. In terpolyesters, 3HV ranged from 9 to 32 mol% depending on the valerate concentration and strain used. A maximal terpolyester P(3HB-co-3HV-co-3HHx) content in dry cells was 71 wt%. Transmission electron microscopy study of A. hydrophila 4AK4 harboring phaPCJ revealed the full occupation of terpolyester P(3HB-co-3HV-co-HHx) in the cellular spaces. Terpolyesters with various monomer compositions showed changing thermal and mechanical properties. Those with higher 3HV fractions demonstrated an improved property over the lower HV containing ones.