Production of Poly(3-hydroxyalkanoate)s by Pseudomonas putida Cultivated in a Glycerol/Nonanoic Acid-Containing Medium

The biosynthesis of poly(3-hydroxyalkanoate) (PHA) by Pseudomonas putida (JCM6160) cultivated in a medium containing glycerol, nonanoic acid, or a glycerol/nonanoic acid mixture as the sole carbon sources was investigated. The PHA content was ~20 % when glycerol was the carbon source. This relatively low content can be attributed to the glycerol end-cap effect and the absence of enzymes that can directly synthesize PHA from acetyl CoA, which is the major metabolite of glycerol. Fatty acids, containing even numbered carbons, are synthesized from acetyl CoA, and they can be used as substrates for PHA synthesis. However, this process also results in decreasing PHA content as fatty acids are siphoned off into other pathways. However, addition of 5 mM nonanoic acid into a 20 mM glycerol-containing medium dramatically increased the PHA content in P. putida, which was 1.3 times larger than the sum of the values found when glycerol and nonanoic acid were each used as the sole carbon source. The PHA, synthesized in the glycerol/nonanoic acid medium, contains 3-hydroxy alkanoate units that have 5, 6, 7, 8, 9, or 10 carbons. The units that contain the even numbered carbons are derived from fatty acids that were produced from glycerol; whereas, the PHA units with the odd numbered carbons are derived from nonanoic acid. Pentanoate units were also found in the polyester derived from glycerol and nonanoic acid, and must have been synthesized indirectly via β-oxidation of nonanoic acid with the assistance of glycerol because pentanoate units were not found in PHA when P. putida was cultivated in the presence of only nonanoic acid.     

Influence of Short Central PEO Segment on Hydrolytic and Enzymatic Degradation of Triblock PCL Copolymers

Hydrolytic, enzymatic degradation and composting under controlled conditions of series of triblock PCL/PEO copolymers, PCEC, with central short PEO block (M _n 400 g/mol) are presented and compared with homopolymer (PCL). The PCEC copolymers, synthesized via ring-opening polymerization of ε-caprolactone, were characterized by ¹H NMR, quantitative ¹³C NMR, GPC, DSC and WAXS. The introduction of the PEO central segment (<?2 wt%) in PCL chains significantly affected thermal degradation and crystallization behavior, while the hydrophobicity was slightly reduced as confirmed by water absorption and moisture uptake experiments. Hydrolytic degradation studies in phosphate buffer after 8 weeks indicated a small weight loss, while FTIR analysis detected changes in crystallinity indexes and GPC measurements revealed bulk degradation. Enzymatic degradation tested by cell-free extracts containing Pseudomonas aeruginosa PAO1 confirmed high enzyme activity throughout the surface causing morphological changes detected by optical microscopy and AFM analysis. The changes in roughness of polymer films revealed surface erosion mechanism of enzymatic degradation. Copolymer with the highest content of PEO segment and the lowest molecular weight showed better degradation ability compared to PCL and other copolymers. Furthermore, composting of polymer films in a model compost system at 37 °C resulted in significant degradation of the all synthesized block copolymers.     

Poly(vinyl alcohol-co-lactic acid)/Hydroxyapatite Composites: Synthesis and Characterization

There is a wide range of applications where calcium phosphate and hydroxyapatite (HA) are used as biomaterials, e.g. as synthetic bone grafts, coating on metal prostheses (like hip endoprostheses or dental implants) and drug carriers. In the study, the design and synthesis of composites based on poly(vinyl alcohol-co-lactic acid)/hydroxyapatite (PVA-co-LA/HA) with potential for biomedical applications, they are presented. The hydroxyapatite particles were surface-grafted with l(+)-lactic acid in the presence of manganese acetate as catalyst, resulting in modified hydroxyapatite (HA_m) with improved capacity of bonding, respectively for the preparation of the composite based on PVA-co-LA/HA_m. FT-IR spectra further confirmed the existence of PLA polymer on the surface of HA particles. In synthesis of PVA-co-LA copolymer the different molar ratios PVA/LA (2/1, 1/1, 1/2), toluene/water: 1/2 (as azeotrope solvent mixture) and manganese acetate as catalyst, were used. The composite materials were synthesized in situ with 10 wt% HA, and respectively HA_m (reported to PVA and lactic acid components). The composite materials were characterized by FTIR spectroscopy, thermal analyses (DSC, DTG), ¹H-NMR spectroscopy, particle size distribution and zeta potential.     

Adsorption of Crystal Violet Dye from Aqueous Solution by Poly(Acrylamide-<Emphasis Type="Italic">co</Emphasis>-Maleic Acid)/Montmorillonite Nanocomposite

Poly(acrylamide-co-maleic acid)/montmorillonite nanocomposites, were synthesized via in situ polymerization with different maleic acid and MMT content. The capability of the hydrogel for adsorption of crystal violet (CV) was investigated in aqueous solutions at different pH values and temperatures. The pseudo-second-order kinetics model could fit successfully the adsorption kinetic data. The effects of maleic acid to acrylamide molar ratio (MA_R), weight percent of MMT (MMT%), the pH of medium and the solution temperature (T) on the CV adsorption capacity (q _e ) of adsorbents were studied by Taguchi experimental design approach. The results indicated that increasing the MMT% leads to a greater q _e . The q _e value of adsorbents increased also with increasing both MA_R and pH, while reduced when the temperature of medium increased. The relatively optimum conditions to achieve a maximum CV adsorption capacity for P(AAm/MA)/MMT adsorbents were found as: 0.06 for MA_R and 5 % of MMT%, medium pH = 7 and T = 20 °C.     

Enhancement of biogas production potential from <Emphasis Type="Italic">Acacia</Emphasis> leaf waste using alkaline pre-treatment and co-digestion

The objective of this research was to evaluate possibility of utilizing Acacia leaves (A. mangium and A. auriculiformis), which is an agro-industrial waste from the pulp and paper industry. The effects of alkaline pre-treatment and co-digestion with Napier grass for the enhancement of biogas production from Acacia leaf waste (ALW) were investigated. Six continuous stirred tank reactors with a working volume of 5 L were carried out at the laboratory scale. The results showed that pre-treatment of Acacia leaf waste (pretreated ALW) by soaking in 3 % NaOH for 48 h increased the biogas and methane productivity to 0.200 and 0.117 m³/kgVS_added compared to 0.098 and 0.048 m³/kgVS_added of raw ALW digestion, respectively. Meanwhile, the co-digestion of Acacia leaves with different proportions of Napier grass at ratios of 1:1–1:3 in volatile solid basis also increased the production of biogas and its productivity. The maximum gas production yields of 0.424 and 0.268 m³/kgVS_added for biogas and methane were obtained at 1:3 ratio. This finding affirms the potential of ALW and its possibility to use as biogas feedstock in both single and co-substrate with Napier grass.     

Water Absorption Kinetics and Hygrothermal Aging of Poly(lactic acid) Containing Halloysite Nanoclay and Maleated Rubber

Poly(lactic acid)/halloysite nanoclay composites (PLA/HNC) containing maleic anhydride grafted styrene-ethylene/butylene-styrene (SEBS-g-MAH) were produced using melt compounding followed by compression molding. The effects of hygrothermal aging on the thermal properties and functional groups changes of the HNC reinforced PLA (with and without SEBS-g-MAH) at three different temperatures (i.e., 30, 40 and 50 °C) were analyzed using differential scanning calorimetry and Fourier transform infrared spectroscopy techniques. The diffusion coefficient (D) of PLA was decreased by the incorporation of HNC and SEBS-g-MAH. The activation energy of water diffusion (E _a ) of PLA/HNC/SEBS-g-MAH nanocomposites was higher than that of pure PLA. The glass transition temperature (T _g ), cold-crystallization temperature (T _cc ) and melting temperature (T _m ) of the PLA sample were shifted to lower temperature and the effect was more pronounced at 50 °C. The carbonyl index values of all PLA samples increased after immersed in 40 and 50 °C, which is due to the formation of higher amount of carboxyl groups during the hydrolysis process.     

Kinetic Analysis of the Temperature Effect on Polyhydroxyalkanoate Production by <Emphasis Type="Italic">Haloferax mediterranei</Emphasis> in Synthetic Molasses Wastewater

Haloferax mediterranei is an extremely halophilic archaeon that is able to synthesize polyhydroxyalkanoate (PHA) in high salt environment with low sterility demand. In this study, a mathematical model was validated and calibrated for describing the kinetic behavior of H. mediterranei at 15, 20, 25, and 35 °C in synthetic molasses wastewater. Results showed that the production of PHA by H. mediterranei, ranging from 390 to 620 mg h^?1 L^?1, was strongly dependent on the temperature. The specific growth rate (µ _max), specific substrate utilization rate (q _max), and specific decay rate (k _d) of H. mediterranei increased with temperature following Arrhenius equation prediction. The estimated activation energy was 58.31, 25.59, and 22.38 kJ mol^?1 for the process of cell growth, substrate utilization, and cell decay of H. mediterranei, respectively. The high temperature triggered the increased PHA storage even without nitrogen limitation. Thus, working at high temperatures seems a good strategy for improving the PHA productivity of H. mediterranei.     

A Microscopic Study of Paper Decayed by <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> and <Emphasis Type="Italic">Paecilomyces variotii</Emphasis>

The current study is interested in evaluating the decay of cotton, Whatman and chemical pulp caused by Trichoderma harzianum and Paecilomyces variotii. The structural changes of the paper were evaluated by Infrared Spectroscopy (FTIR) and Scanning Electron Microscope (SEM). The SEM results show differences in hyphae colonization and paper decay patterns between studied species under the current study; P. variotii caused an eroded structure in the cotton (cavity forming), whereas the initial T. harzianum colonization produced rupture and erosion (soft-rot decay type II) for the three types of paper ,the gaps were elongated with sharp pointed ends, which consisted either of individual cavities or in chains. Moreover, FTIR results confirmed that there a relationship could be observed between fungal decay and crystalline cellulose content because the intensity of peaks at 1335 and 1111 cm^?1 significantly decreased due to the fungal decay. Furthermore, the intensity of O–H stretching absorption slightly decreased, and this may be attributed to hydrolysis of cellulose molecules.     

Investigation of Biosurfactant Activity and Asphaltene Biodegradation by <Emphasis Type="Italic">Bacillus cereus</Emphasis>

In this research, a biosurfactant-producing bacterium with capability of asphaltene degradation was isolated from oil-contaminated soil samples, and identified as Bacillus cereus. This strain produced an effective biosurfactant in the presence of molasses and the surface tension was reduced to the level of 36.4 mN/m after 48 h under optimum conditions. The optimum values of carbon-to-nitrogen ratio (C:N), pH, and temperature for biosurfactant production were determined as 30:1, 7.3 and 29 °C, respectively, using response surface methodology. The maximum emulsification activity in the culture broth was 53 % after 48 h using kerosene at 25 °C. The goodness of fit of four growth kinetic models including Tessier, Contois, Logistic and Westerhoff was compared for the bacterial growth and molasses utilization of B. cereus in 5-L batch bioreactor during 120 h. Conducted kinetic study showed that biosurfactant production had a good fit with the Contois growth kinetic model (R² = 0.962) and the maximum specific growth rate (µ _max ), saturation constant (K _s ) and the yield of biomass per substrate (Y _x/s ) were determined to be 0.145 h^?1, 1.83 g/L and 0.428 g/g, respectively. The asphaltene biodegradation in flask was evaluated by FTIR analysis and quantified by a spectrophotometer. This bacterium was able to degrade up to 40 % of asphaltene as a sole carbon and energy source after 60 days at 28 °C. The resulting surface tension of 30.2 mN/m with the critical micelle concentration of 23.4 mg/L indicated good efficiency of the biosurfactant.     

Biodegradation of Natural Rubber and Natural Rubber Products by <Emphasis Type="Italic">Streptomyces</Emphasis> sp. Strain CFMR 7

The rubber degrading activity of Streptomyces sp. CFMR 7 whose whole genome sequence was recently determined was tested with non-vulcanized fresh latex and common vulcanized rubber products such as latex glove, latex condom and latex car tyre. The degradation activity was unequivocally demonstrated by scanning electron microscopy with respect to microbial colonization efficiency, disintegration of rubber material and biofilm formation after 3, 6 and 9 months of inoculation. Fourier transform infrared spectroscopy comprising the attenuated total reflectance analysis on these inoculated products revealed insights into the biodegradation mechanism of this strain whereby, a decrease in the number of cis -1,4 double bonds in the polyisoprene chain, the appearance of ketone and aldehyde groups formation indicating an oxidative attack at the double bond of rubber hydrocarbon. In the presence of strain Streptomyces sp. CFMR 7, gel permeation chromatography analysis revealed a significant shift of the molecular weight distribution to lower values. Clear decrease in the molecular weight was observed over 3, 6 and 9 months of cultivation on fresh latex samples compared to other vulcanized products. No shift in the molecular weight distribution was observed for non-inoculated control. These results clearly showed that Streptomyces sp. CFMR 7 was able to cleave the carbon backbone of poly (cis -1,4-isoprene). Although this strain was able to degrade both non-vulcanized and vulcanized rubber products, faster degradation was obtained with natural rubber and rubber products with low complexity.     

Degradation Mechanism of CF/EP Composites in Supercritical <Emphasis Type="Italic">n</Emphasis>-Butanol with Alkali Additives

CF/EP (carbon fibre/epoxy resin) composites were degraded by supercritical n-butanol with alkali additive KOH in a batch reactor. The catalytic degradation mechanism of the composites was investigated based on the analysis of liquid phase products by GC–MS and solid phase products by FTIR. The results indicate that alkali additive (KOH) can promote Guerbet reaction and increase hydrogen donor capability of supercritical n-butanol. The H^· can combine promptly with the free radical formed by the scission of linear and crosslinked chains in epoxy resin to generate the liquid products, including phenol, 4-isopropylphenol, 4-(2-methylallyl)phenol and other derivatives of benzene and phenol. The combination of supercritical n-butanol with alkali additive is an effective way to degrade and recycle CF/EP composites.     

Experimental Investigation of Cellulose/Silver Nanocomposites Using In Situ Generation Method

Cellulose gel films were prepared by regeneration process using pre-cooled aq.(8 wt% LiOH + 15 wt% urea) mixture as solvent and ethyl alcohol as non solvent. The Terminus cattapa leaf extract diffused wet cellulose films were then dipped in 1–5 mM aq.AgNO₃ solutions to allow in situ generation of silver nanoparticles (AgNPs). Besides the in situ generation, some AgNPs were also formed outside the wet films in the solution. The AgNPs formed outside the films were observed under transmission electron microscope and scanning electron microscope. The nanocomposite films were also characterized by Fourier transform infrared spectroscopy, X-ray diffraction and thermogravimetric analysis and tensile test. The thermal stability of the composite films was lower than that of the matrix up to a temperature of ~300 °C and afterwards showed a reverse trend. The tensile strength of the nanocomposite films was found to be higher than the matrix but decreased with increasing concentration of aq.AgNO₃. The cellulose/AgNPs composite films showed good antibacterial activity against E. coli (gram positive) and Bacillus sp. (gram negative). Based on the aforementioned properties, the cellulose/AgNPs composite films can be considered for antibacterial packaging and medical applications.     

In Vivo and In Vitro Degradation Studies for Poly(3-hydroxybutyrate-<Emphasis Type="Italic">co</Emphasis>-3-hydroxyhexanoate) Biopolymer

Studies have shown that the copolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(HB-co-HHx)] possesses favorable mechanical properties for use in medical supplies and products (e.g., sutures, scaffolds, bone plates). One of the major under-addressed issues associated with the use of biodegradable, bio-based PHA polymers in resorbable medical products is the correlation between the mechanical properties and the in vivo material degradation over time. In this study, P(HB-co-17 mol% HHx) matrices were mechanically tested after either incubation in cultures of human embryonic kidney cells (HEK) for in vitro degradation studies for up to 4 weeks, or inserted into Danio rerio (zebrafish) tissues for in vivo degradation studies for up to 7 weeks. The mechanical properties and scanning electron microscopy (SEM) images of the degraded materials were examined and later correlated to understand the degradation phenomenon. Our results show that Young’s modulus of P(HB-co-17 mol%HHx) during in vitro studies decreased from 3.26 to 2.42 GPa within 4 weeks, and in vivo breakdown resulted in a significant decrease in Young’s modulus with a decrease from 3.26 to 0.51 GPa and a mass loss of 59 % within 7 weeks. SEM images showed the development of pores and cracks on the surface of the material over time. Plasticization and recrystallization were observed and likely play a role in the alteration of mechanical properties.     

Synthesis of poly(hydroxyalkanoate) from hydrolyzed linseed oil

Pseudomonas putida was grown on a mixture of long-chain fatty acids obtained by hydrolysis of linseed oil. A poly(3-hydroxyalkanoate) containing 51.2% of unsaturated monomers was obtained. A considerable percentage (13.6%) was constituted by C₁₄ and C₁₆ monomers containing three double-bonds in the side chains. The polymer showed a high tendency to crosslink when it was kept in presence of air. In the crosslinked polymer no polyunsaturated monomers could be detected.     

Influence of TiO<Subscript>2</Subscript> Nanoparticles on the Crystallization Behaviour and Tensile Properties of Biodegradable PLA and PCL Nanocomposites

This paper investigated the influence of TiO₂ nanoparticles on the morphologies, as well as crystallization behaviour and kinetics, of neat PLA and PCL, and of these polymers in different PLA/PCL blends. We used transmission electron microscopy to evaluate the morphologies of the systems, while the crystallization behaviour and kinetics were investigated through differential scanning calorimetry (DSC). In addition to standard and modulated (StepScan) DSC analyses, the self-nucleation temperatures of neat PCL and PCL in the different nanocomposites were determined, followed by a self-nucleation and annealing thermal fractionation analysis of PCL crystallization and an Avrami isothermal kinetic analysis of PCL crystallization and PLA cold crystallization. We found that the nanoparticles were well dispersed, but only in the PLA phase of the blends, with only a few on the interface or in the PCL phase. They did nucleate and accelerate, and influence the mechanism of, the PCL crystallization in neat PCL, but had little influence on PCL crystallization in the blends. They strongly influenced the rate of cold crystallization of PLA, but had little influence on this parameter in the blends. The tensile properties were also determined, and changes in these properties could be related to the morphologies of the systems.     

Preparation and characterization of high surface area activated carbon from pine wood sawdust by fast activation with H<Subscript>3</Subscript>PO<Subscript>4</Subscript> in a spouted bed

Activated carbons were produced from waste pine wood sawdust using fast activation with H₃PO₄ in a spouted bed. In this study, activation temperature was set as 800 °C, and activation time ranged from 1 to 15 min. Experimental results show that sawdust impregnated with higher mass ratio of H₃PO₄ would be agglomerated in spouted bed, and difficult to fluidize. Therefore, an amount of quartz sand was added to assist for good fluidization. Fluidization of particle can improve the BET surface area or micropore volume of activated carbons. High BET surface area activated carbons can be obtained with activation time of only 1–5 min by combining the fluidization and H₃PO₄ fast activation. The obtained activated carbons contained developed pore structure and abundant surface functional groups (carboxyl, carbonyl and P-containing groups) by SEM–EDS, FTIR and XPS techniques. The particles of impregnation ratio of 1:1 can achieve fluidization without adding the quartz sand, which was convenient for experimental operation and even industrial production, and the BET surface area can reach more than 1000 m²/g in activation time of only 5 min.     

Itaconic Acid Grafted Starch Hydrogels as Metal Remover: Capacity,Selectivity and Adsorption Kinetics

Hydrogels were synthesized by free radical graft copolymerization of itaconic acid (IA) onto corn starch (S-g-IA). For this purpose, potassium permanganate (KMnO₄)-sodium bisulfite (NaHSO₃) was used as redox initiation system. The formation of grafted starches was confirmed by Fourier transform infrared spectroscopy, wide angle X-ray scattering, thermogravimetric analysis and scanning electron microscopy. The effect of monomer concentration, neutralization, addition of crosslinking agent, N,N-bismetilenacrilamide (MBAm), and initiator concentration on grafting efficiency and adsorption capacity of the starch hydrogels was investigated. It was demonstrated that the introduction of carboxyl and carbonyl groups promoted starch hydration and swelling. Grafting degree increased with the decrease of monomer concentration, increase of initiator concentration, grade of neutralization and the addition of MBAm without neutralization. Remarkably the resulting materials exhibited water absorption capacities between 258 and 1878% and the ability to adsorb metal ions. It was experimentally confirmed the metal uptake, obtaining the higher adsorption capacity (q _e  = 35 mg/g) for the product prepared with the pre-oxidation and lower initiator concentration. The removal capacity order was Pb²⁺>Ni²⁺>Zn²⁺>Cd²⁺. Moreover, the experimental kinetic and the equilibrium adsorption data for Ni²⁺ and Pb²⁺ were best fitted to the pseudo-second order and Freundlich isotherm models, respectively. This work describes for the first time the preparation of metal removal hydrogels based on starch and itaconic acid using the pair redox system KMnO₄/NaHSO₃, which avoids the starch hydrolysis and allows itaconic acid grafting incorporation without the requirement of more reactive comonomers.     

Preparation and Characterization of Polyaniline and Ag/ Polyaniline Composite Nanoporous Particles and Their Antimicrobial Activities

Polyaniline (PANI) and Ag/PANI nanoporous composite were prepared by an oxidative polymerization method. The oxidation process of PANI nanoparticles was occurred using (NH₄)₂S₂O₈ while the oxidation process of Ag/PANI nanoporous composite was occurred using AgNO₃ under the effect of artificial radiation. The structural, morphological, and optical properties of the PANI and Ag/PANI nanoporous structures were studied using different characterization tools. The results confirm the formation of polycrystalline nanoporous PANI and spherical nanoporous composite of Ag/PANI particles. Antibacterial activity tests against gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus, and gram-negative bacteria, Escherichia coli, and Salmonella species were carried out using different concentrations of PANI nanoparticles and Ag/PANI nanoporous composites. PANI has not antibacterial effect against all studied pathogens. In contrast, Ag/PANI nanoporous composites possessed antibacterial activity that is identified by the zone of inhibition. The inhibition zones of bacteria are in order; Salmonella species?>?S. aureus?>?B. subtilis?>?E. coli. The inhibition zones of all bacteria increased with increasing concentrations of Ag/PANI nanoporous composites from 200 to 400 ppm then decreased with further increasing of the dose concentrations to 600 ppm. Finally, a simplified mechanism based on the electrostatic attraction is presented to describe the antimicrobial activity of Ag/PANI nanoporous composite.     

Preparation and Characterization of Collagens from the Skin of Northern Snakehead (<Emphasis Type="Italic">Channa argus)</Emphasis>

Channa argus, a type of snakehead fish native to China, is a popular food fish in certain Asian countries but is a known destructive invasive species in the US. In this study, the two collagens, i.e. acid-soluble collagen (ASC) and pepsin-solubilized collagen (PSC), were obtained from C. argus skin. The yield of ASC was 28.0% and that of PSC was 16.8% on the dry bases. The collagens were identified as the collagen of type I by SDS–PAGE patterns. The Tds were approximately 27.0?°C. Similar ultraviolet spectra of both collagens were observed. Fourier Transform infrared spectra indicated PSC structure had a little change due to the loss of terminal domains by pepsin digestion. The results of XRD proved that the two collagens retained their helical structures. The results suggest that the collagens isolated from C. argus can potentially be alternative sources of vertebrate collagens for use in the food and other industries.     

Increased 3HV Concentration in the Bacterial Production of 3-Hydroxybutyrate (3HB) and 3-Hydroxyvalerate (3HV) Copolymer with Acid-Digested Rice Straw Waste

Bacterial synthesis of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) copolymer [P(3HB-co-3HV)] using the hydrolysate of rice straw waste as a carbon source was affected by the composition of the hydrolysate, which depends highly on the rice straw pretreatment condition. Acid digestion with 2 % sulfuric acid generated larger production of P(3HB-co-3HV) than 6 % sulfuric acid, but 3HV concentration in the copolymer produced with 2 % acid hydrolysate was only 8.8 % compared to 18.1 % with 6 % acid hydrolysate. To obtain a higher 3HV mole fraction for enhanced flexibility of the copolymer, an additional heating was conducted with the 2 % acid hydrolysate after removal of residual rice straw. As the additional heating time increased a higher concentration of levulinic acid was generated, and consequently, the mole fraction of 3HV in P(3HB-co-3HV) increased. Among the conditions tested (i.e., 20-, 40-, 60-min), 60-min additional heating following 2 % sulfuric acid digestion achieved the highest 3HV mole fraction of 22.9 %. However, a longer heating time decreased the P(3HB-co-3HV) productivity, probably due to the increased intermediates concentrations acting as inhibitors in the hydrolysates. Therefore, the use of additional heating needs to consider both the increase in the 3HV mole fraction and the decrease in the P(3HB-co-3HV) productivity.