Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer is noted for its high biocompatibility, which makes it an excellent candidate for biopharmaceutical applications. The wild-type Cupriavidus sp. USMAA1020 strain is able to synthesize P(3HB-co-4HB) copolymers with different 4HB monomer compositions (up to 70mol%) in shaken flask cultures. Combinations of 4HB carbon precursors consisting of 1,6-hexanediol and γ-butyrolactone were applied for the production of P(3HB-co-4HB) with different 4HB molar fraction. A sharp increase in 4HB monomer composition was attained by introducing additional copies of PHA synthase gene (phaC), responsible for P(3HB-co-4HB) polymerization. The phaC of Cupriavidus sp. USMAA1020 and Cupriavidus sp. USMAA2-4 were cloned and heterologously introduced into host, wild-type Cupriavidus sp. USMAA1020. The gene dosage treatment resulted in the accumulation of 93mol% 4HB by the transformant strains when grown in similar conditions as the wild-type USMAA1020. The PHA synthase activities for both transformants were almost two-fold higher than the wild-type. The ability of the transformants to produce copolymers with high 4HB monomer composition was also tested in large scale production system using 5L and 30L bioreactors with a constant oxygen mass transfer rate. The 4HB monomer composition could be maintained at a range of 83-89mol%. The mechanical and thermal properties of copolymers improved with increasing 4HB monomer composition. The copolymers produced could be tailored for specific biopharmaceutical applications based on their properties.
This study reports the production of P(3HB-co-4HB) [Poly(3-hydroxybutyrate-co-4-hydroxybutyrate)] in possession of high molecular weight and elastomeric properties by Cupriavidus sp. USMAA1020 in single-stage mixed-substrate cultivation system. 1,4-butanediol and 1,6-hexanediol are found to be efficient substrate mixture that has resulted in high copolymer yield, occupying a maximum of 70wt% of the total biomass and producing higher 4HB monomer composition ranging from 31mol% to 41mol%. In substrate mixtures involving 1,6-hexanediol, cleavage of the 6-hydroxyhexanoyl-CoA produces Acetyl-CoA and 4-hydroxybutyryl-CoA. Acetyl-CoA is instrumental in initiating the cell growth in the single-stage fermentation system, preventing 4-hydroxybutyryl-CoA from being utilized via β-oxidation and retained the 4HB monomer at higher ratios. Macroscopic kinetic models of the bioprocesses have revealed that the P(3HB-co-4HB) formation appears to be in the nature of mixed-growth associated with higher formation rate during exponential growth phase; evidenced by higher growth associated constants, α, from 0.0690g/g to 0.4615g/g compared to non-growth associated constants, β, from 0.0092g/g/h to 0.0459g/g/h. The P(3HB-co-31mol% 4HB) produced from the substrate mixture exhibited high weight-average molecular weight, Mwof 927kDa approaching a million Dalton, and possessed elongation at break of 1637% upon cultivation at 0.56wt% C. This is the first report on such properties for the P(3HB-co-4HB) copolymer. The copolymer is highly resistant to polymer deformation after being stretched.
A statistical approach with D-optimal design was used to optimize the process parameters for polycaprolactone (PCL) synthesis. The variables selected were temperature (50°C-110°C), time (1-7 h), mixing speed (50-500 rpm) and monomer/solvent ratio (1:1-1:6). Molecular weight was chosen as response and was determined using matrix-assisted laser desorption/ionization time of flight (MALDI TOF). Using the D-optimal method in design of experiments, the interactions between parameters and responses were analysed and validated. The results show a good agreement with a minimum error between the actual and predicted values.
Modified rice straw/Fe3O4/polycaprolactone nanocomposites (ORS/Fe3O4/ PCL-NCs) have been prepared for the first time using a solution casting method. The RS/Fe3O4-NCs were modified with octadecylamine (ODA) as an organic modifier. The prepared NCs were characterized by using X-ray powder diffraction (XRD), Scanning electron microscopy (SEM), Transmission electron microscopy (TEM), Thermogravimetric analysis (TGA) and Fourier transform infrared spectroscopy (FT-IR). The XRD results showed that as the intensity of the peaks decreased with the increase of ORS/Fe3O4-NCs content in comparison with PCL peaks, the Fe3O4-NPs peaks increased from 1.0 to 60.0 wt. %. The TEM and SEM results showed a good dispersion of ORS/Fe3O4-NCs in the PCL matrix and the spherical shape of the NPs. The TGA analysis indicated thermal stability of ORS/Fe3O4-NCs increased after incorporation with PCL but the thermal stability of ORS/Fe3O4/PCL-NCs decreased with the increase of ORS/Fe3O4-NCs content. Tensile strength was improved with the addition of 5.0 wt. % of ORS/Fe3O4-NCs. The antibacterial activities of the ORS/Fe3O4/PCL-NC films were examined against Gram-negative bacteria (Escherichia coli) and Gram-positive bacteria (Staphylococcus aureus) by diffusion method using nutrient agar. The results indicated that ORS/Fe3O4/PCL-NC films possessed a strong antibacterial activity with the increase in the percentage of ORS/Fe3O4-NCs in the PCL.
The medium-chain-length polyhydroxyalkanoate (PHA(MCL)) produced by Pseudomonas putida PGA1 using saponified palm kernel oil as the carbon source could degrade readily in water taken from Kayu Ara River in Selangor, Malaysia. A weight loss of 71.3% of the PHA film occurred in 86 d. The pH of the river water medium fell from 7.5 (at d 0) to 4.7 (at d 86), and there was a net release of CO2. In sterilized river water, the PHA film also lost weight and the pH of the water fell, but to lesser extents. The C8 monomer of the PHA was completely removed after 6 d of immersion in the river water, while the proportions of the other monomers (C10, C12, and C14) were reversed from that of the undegraded PHA. By contrast, the monomer composition of the PHA immersed in sterilized river water did not change significantly from that of the undegraded PHA. Scanning electron microscopy showed physical signs of degradation on the PHA film immersed in the river water, but the film immersed in sterilized river water was relatively unblemished. The results thus indicate that the PHA(MCL) was degraded in tropical river water by biologic as well as nonbiologic means. A significant finding is that shorter-chain monomers were selectively removed throughout the entire PHA molecule, and this suggests enzymatic action.
Burkholderia sp. synthase has been shown to polymerize 3-hydroxybutyrate (3HB), 3-hydroxyvalerate, and 3-hydroxy-4-pentenoic acid monomers. This study was carried out to evaluate the ability of Burkholderia sp. USM (JCM 15050) and its transformant harboring the polyhydroxyalkanoate (PHA) synthase gene of Aeromonas caviae to incorporate the newly reported 3-hydroxy-4-methylvalerate (3H4MV) monomer. Various culture parameters such as concentrations of nutrient rich medium, fructose and 4-methylvaleric acid as well as harvesting time were manipulated to produce P(3HB-co-3H4MV) with different 3H4MV compositions. The structural properties of PHA containing 3H4MV monomer were investigated by using nuclear magnetic resonance and Fourier transform infrared spectroscopy (FTIR). The relative intensities of the bands at 1,183 and 1,228 cm⁻¹ in the FTIR spectra enabled the rapid detection and differentiation of P(3HB-co-3H4MV) from other types of PHA. In addition, the presence of 3H4MV units in the copolymer was found to considerably lower the melting temperature and enthalpy of fusion values compared with poly(3-hydroxybutyrate) (P(3HB)). The copolymer exhibited higher thermo-degradation temperature but similar molecular weight and polydispersity compared with P(3HB).
Chitosan, collagen, gelatin, polylactic acid and polyhydroxyalkanoates are notable examples of biopolymers, which are essentially bio-derived polymers produced by living cells. With the right techniques, these biological macromolecules can be exploited for nanotechnological advents, including for the fabrication of nanocarriers. In the world of nanotechnology, it is highly essential (and optimal) for nanocarriers to be biocompatible, biodegradable and non-toxic for safe in vivo applications, including for drug delivery, cancer immunotherapy, tissue engineering, gene delivery, photodynamic therapy and many more. The recent advancements in understanding nanotechnology and the physicochemical properties of biopolymers allows us to modify biological macromolecules and use them in a multitude of fields, most notably for clinical and therapeutic applications. By utilizing chitosan, collagen, gelatin, polylactic acid, polyhydroxyalkanoates and various other biopolymers as synthesis ingredients, the 'optimal' properties of a nanocarrier can easily be attained. With emphasis on the aforementioned biological macromolecules, this review presents the various biopolymers utilized for nanocarrier synthesis along with their specific synthetization methods. We further discussed on the characterization techniques and related applications for the synthesized nanocarriers.
An obligate anaerobic bacterium Clostridium difficile has a unique metabolic pathway to convert leucine to 4-methylvalerate, in which 4-methyl-2-pentenoyl-CoA (4M2PE-CoA) is an intermediate of this pathway. 4M2PE-CoA is also able to be converted to 3-hydroxy-4-methylvalerate (3H4MV), a branched side chain monomer unit, for synthesis of polyhydroxyalkanoate (PHA) copolymer. In this study, to synthesize 3H4MV-containing PHA copolymer from leucine, the leucine metabolism-related enzymes (LdhA and HadAIBC) derived from C. difficile and PHA biosynthesis enzymes (PhaPCJAc and PhaABRe) derived from Aeromonas caviae and Ralstonia eutropha were co-expressed in the codon usage-improved Escherichia coli. Under microaerobic culture conditions, this E. coli was able to synthesize P(3HB-co-12.2 mol% 3H4MV) from glucose with the supplementation of 1 g/L leucine. This strain also produced P(3HB-co-12.6 mol% 3H4MV) using the culture supernatant of leucine overproducer E. coli strain NS1391 as the medium for PHA production, achieving 3H4MV copolymer synthesis only from glucose. Furthermore, we tested the feasibility of the 3H4MV copolymer synthesis in E. coli strain NS1391 from glucose. The recombinant E. coli NS1391 was able to synthesize P(3HB-co-3.0 mol% 3H4MV) from glucose without any leucine supplementation. This study demonstrates the potential of the new metabolic pathway for 3H4MV synthesis using leucine metabolism-related enzymes from C. difficile.
In this work, polylactic acid (PLA) reinforced cellulose nanowhiskers (CNW) were prepared through solution casting technique. The CNW was first isolated from oil palm empty fruit bunch microcrystalline cellulose (OPEFB-MCC) by using 64% H2SO4 and was designated as CNW-S. The optical microscopy revealed that the large particle of OPEFB-MCC has been broken down by the hydrolysis treatment. The atomic force microscopy confirmed that the CNW-S obtained is in nanoscale dimension and appeared in individual rod-like character. The produced CNW-S was then incorporated with PLA at 1, 3, and 5 parts per hundred (phr) resins for the PLA-CNW-S nanocomposite production. The synthesized nanocomposites were then characterized by a mean of tensile properties and thermal stability. Interestingly to note that incorporating of 3 phr/CNW-S in PLA improved the tensile strength by 61%. Also, CNW-S loading showed a positive impact on the Young's modulus of PLA. The elongation at break (Eb) of nanocomposites, however, decreased with the addition of CNW-S. Field emission scanning electron microscopy and transmission electron microscopy revealed that the CNW-S dispersed well in PLA at lower filler loading before it started to agglomerate at higher CNW-S loading (5phr). The DSC analysis of the nanocomposites obtained showed that Tg,Tcc and Tm values of PLA were improved with CNW-S loading. The TGA analysis however, revealed that incopreated CNW-S in PLA effect the thermal stability (T10,T50 and Tmax) of nanocomposite, where it decrease linearly with CNW-S loading.
The scope of additive manufacturing, particularly fused deposition modelling (FDM), can indeed be explored with the fabrication of multi-material composite laminates using this technology. Laminar composite structures made up of two distinct materials, namely acrylonitrile butadiene styrene (ABS) and carbon fiber reinforced polylactic acid (CF-PLA), were produced using the FDM process. The current study analyzes the effect of various printing parameters on the interfacial bond strength (IFBS) of the ABS/CF-PLA laminar composite by employing response surface methodology. The physical examination of the tested specimens revealed two failure modes, where failure mode 1 possessed high IFBS owing to the phenomenon of material patch transfer. Contrarily, failure mode 2 yielded low IFBS, while no patch transfer was observed. The analysis of variance (ANOVA) revealed that printing parameters were highly interactive in nature. After extensive experimentation, it was revealed that good quality of IFBS is attributed to the medium range of printing speed, high infill density, and low layer height. At the same time, a maximum IFBS of 20.5 MPa was achieved. The study presented an empirical relation between printing parameters and IFBS that can help in forecasting IFBS at any given printing parameters. Finally, the optimized printing conditions were also determined with the aim to maximize IFBS.
This study was aimed at fabricating composites of polylactic acid (PLA) matrix-reinforced oil palm empty fruit bunch (OPEFB) fiber filled with chemically reduced graphene oxide (rGO). A total of 2-8 wt.% rGO/OPEFB/PLA composites were characterized for their complex permittivity using an open-ended coaxial probe (OEC) technique. The shielding efficiency properties were calculated using the measured transmission (S21) and the reflection (S11) coefficient results. All the measurements and calculations were performed in the 8-12 GHz frequency range. The morphological and microstructural study included X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), and Fourier transform infrared spectroscopy (FTIR). The results indicated that the incorporation of rGO as filler into the composites enhanced their complex permittivity properties. The composites showed a total shielding efficiency (SET) of about 31.2 dB at a frequency range of 8-12 GHz, which suggests their usefulness for microwave absorption.
A thermoplastic elastomer (TPE) based nanocomposite with the same weight ratio of hybrid nanofillers composed of carbon nanotubes (CNTs) and montmorillonite nanoclay (DK4) was prepared using a melt blending technique with an internal mixer. The TPE composite was blended from polylactic acid (PLA), liquid natural rubber (LNR) as a compatibilizer and natural rubber (NR) in a volume ratio of 70:10:20, respectively. The weight ratio of CNTs and DK4 was 2.5 wt%. The prepared samples were exposed to gamma radiation at range of 0-250 kGy. After exposure to gamma radiation, the mechanical, thermo-mechanical, thermal and electrical conductivity properties of the composites were significantly higher than unirradiated TPE composites as the irradiation doses increased up to 150 kGy. Transmission electron microscopy (TEM) micrographs revealed the good distribution and interaction between the nano-fillers and the matrix in the prepared TPE hybrid nanocomposites. In summary, the findings from this work definite that gamma irradiation might be a viable treatment to improve the properties of TPE nanocomposite for electronic packaging applications.
Biodegradable polyester polyhydroxyalkanoate (PHA) is a promising bioplastic material for industrial use as a replacement for petroleum-based plastics. PHA synthase PhaC forms an active dimer to polymerize acyl moieties from the substrate acyl-coenzyme A (CoA) into PHA polymers. Here we present the crystal structure of the catalytic domain of PhaC from Chromobacterium sp. USM2, bound to CoA. The structure reveals an asymmetric dimer, in which one protomer adopts an open conformation bound to CoA, whereas the other adopts a closed conformation in a CoA-free form. The open conformation is stabilized by the asymmetric dimerization and enables PhaC to accommodate CoA and also to create the product egress path. The bound CoA molecule has its β-mercaptoethanolamine moiety extended into the active site with the terminal SH group close to active center Cys291, enabling formation of the reaction intermediate by acylation of Cys291.
The nutrition-versatility of Burkholderia sp. strain USM (JCM 15050) has initiated the studies on the use of this bacterium for polyhydroxyalkanoate (PHA) production. To date, the Burkholderia sp. has been reported to synthesize 3-hydroxybutyrate, 3-hydroxyvalerate and 3-hydroxy-4-methylvalerate monomers. In this study, the PHA biosynthetic genes of this strain were successfully cloned and characterized. The PHA biosynthetic cluster of this strain consisted of a PHA synthase (phaC), β-ketothiolase (phaA), acetoacetyl-CoA reductase (phaB) and PHA synthesis regulator (phaR). The translated products of these genes revealed identities to corresponding proteins of Burkholderia vietnamiensis (99-100 %) and Cupriavidus necator H16 (63-89%). Heterologous expression of phaCBs conferred PHA synthesis to the PHA-negative Cupriavidus necator PHB¯4, confirming that phaCBs encoded functionally active protein. PHA synthase activity measurements revealed that the crude extracts of C. necator PHB¯4 transformant showed higher synthase activity (243 U/g) compared to that of wild-types Burkholderia sp. (151 U/g) and C. necator H16 (180 U/g). Interestingly, the transformant C. necator PHB¯4 harbouring Burkholderia sp. PHA synthase gene accumulated poly(3-hydroxybutyrate-co-4-hydroxybutyrate) with 4-hydroxybutyrate monomer as high as up to 87 mol% from sodium 4-hydroxybutyrate. The wild type Burkholderia sp. did not have the ability to produce this copolymer.
In this paper, the influence of processing input parameters on the heat-affected zone (HAZ) of three different material thicknesses of sugar palm fiber reinforced unsaturated polyester (SPF-UPE) composites cut with a CO2 laser was investigated. Laser power, traverse speed, and gas pressure were selected as the most influential input parameters on the HAZ to optimize the HAZ response with fixing all of the other input parameters. Taguchi's method was used to determine the levels of parameters that give the best response to the HAZ. The significance of input parameters was also determined by calculating the max-min variance of the average of the signal-to-noise ratio (S/N) ratio for each parameter. Analysis of variation (ANOVA) was used to determine each input parameter's contribution to the influence on HAZ depth. The general results show that the minimum levels of laser power and the highest levels of traverse speed and gas pressure gave the optimum response to the HAZ. Gas pressure had the most significant effect on the HAZ, with contribution decreases as the material thickness increased, followed by the traverse speed with contribution increases with the increase in material thickness. Laser power came third, with a minimal contribution to the effect on the HAZ, and it did not show a clear relationship with the change in material thickness. By applying the optimum parameters, the desired HAZ depth could be obtained at relatively low values.
The current work is an attempt to reduce friction coefficient of the treated betelnut fibre reinforced polyester (T-BFRP) composites by aging them in twelve different solutions with different kinematic viscosities. The test will be performed on a pin on disc (POD) wear test rig using different applied loads (5⁻30 N), different sliding distances (0⁻6.72 km) at sliding speed of 2.8 m/s subjected to a smooth stainless steel counterface (AISI-304). Different orientations of the fibre mats such as anti-parallel (AP) and parallel (P) orientations subjected to the rotating counterface will be considered. The worn surfaces were examined through optical microscopy imaging and it was found that the aged specimens had significantly lower damages as compared to neat polyester (NP) and the unaged samples. Besides, P-O samples revealed lower friction coefficients as compared to AP-O, i.e., reduction was about 24.71%. Interestingly, aging solutions with lower kinematic viscosities revealed lower friction coefficients of the aged T-BFRP composites when compared to the ones aged in higher kinematic viscosities.
The effectiveness of superheated steam (SHS) as an alternative, eco-friendly treatment method to modify the surface of pineapple leaf fiber (PALF) for biocomposite applications was investigated. The aim of this treatment was to improve the interfacial adhesion between the fiber and the polymer. The treatment was carried out in an SHS oven for different temperatures (190⁻230 °C) and times (30⁻120 min). Biocomposites fabricated from SHS-treated PALFs and polylactic acid (PLA) at a weight ratio of 30:70 were prepared via melt-blending techniques. The mechanical properties, dimensional stability, scanning electron microscopy (SEM), and X-ray diffraction (XRD) for the biocomposites were evaluated. Results showed that treatment at temperature of 220 °C for 60 min gave the optimum tensile properties compared to other treatment temperatures. The tensile, flexural, and impact properties as well as the dimensional stability of the biocomposites were enhanced by the presence of SHS-treated PALF. The SEM analysis showed improvement in the interfacial adhesion between PLA and SHS-treated PALF. XRD analysis showed an increase in the crystallinity with the addition of SHS-PALF. The results suggest that SHS can be used as an environmentally friendly treatment method for the modification of PALF in biocomposite production.
Halloysite nanotubes (HNTs)-polyester nanocomposites with four different concentrations were produced using solution casting technique and the biodegradation effect of short-term seawater exposure (120 h) was studied. Monolithic polyester was observed to have the highest seawater absorption with 1.37%. At 0.3 wt % HNTs reinforcement, the seawater absorption dropped significantly to the lowest value of 0.77% due to increase of liquid diffusion path. For samples tested in dry conditions, the Tg, storage modulus, tensile properties and flexural properties were improved. The highest improvement of Tg was from 79.3 to 82.4 °C (increase 3.1 °C) in the case of 0.3 wt % HNTs. This can be associated with the exfoliated HNTs particles, which restrict the mobility of polymer chains and thus raised the Tg. After seawater exposure, the Tg, storage modulus, tensile properties and flexural properties of polyester and its nanocomposites were decreased. The Young's modulus of 0.3 wt % HNTs-polyester dropped 20% while monolithic polyester dropped up to 24% compared to their values in dry condition. Apart from that, 29% flexural modulus reduction was observed, which was 18% higher than monolithic polyester. In contrast, fracture toughness and surface roughness increased due to plasticization effect. The presence of various microbial communities caused gradual biodegradation on the microstructure of the polyester matrix as also evidently shown by SEM images.
This article describes attenuation and absorption measurements using the microstrip transmission line technique connected with a microwave vector network analyzer (Agilent 8750B). The magnitudes of the reflection (S11) and transmission (S21) coefficients obtained from the microstrip transmission line were used to determine the attenuation and absorption of oil palm empty fruit bunch/polylactic acid (OPEFB/PLA) composites in a frequency range between 0.20 GHz and 12 GHz at room temperature. The main structure of semi-flexible substrates (OPEFF/PLA) was fabricated using different fiber loading content extracted from oil palm empty fruit bunch (OPEFB) trees hosted in polylactic acid (PLA) using the Brabender blending machine, which ensured mixture homogeneity. The commercial software package, Computer Simulation Technology Microwave Studio (CSTMWS), was used to investigate the microstrip line technique performance by simulating and determine the S11 and S21 for microwave substrate materials. Results showed that the materials' transmission, reflection, attenuation, and absorption properties could be controlled by changing the percentage of OPEFB filler in the composites. The highest absorption loss was calculated for the highest percentage of filler (70%) OPEFB at 12 GHz to be 0.763 dB, while the lowest absorption loss was calculated for the lowest percentage of filler 30% OPEFB at 12 GHz to be 0.407 dB. Finally, the simulated and measured results were in excellent agreement, but the environmental conditions slightly altered the results. From the results it is observed that the value of the dielectric constant (εr') and loss factor (εr″) is higher for the OPEFB/PLA composites with a higher content of OPEFB filler. The dielectric constant increased from 2.746 dB to 3.486 dB, while the loss factor increased from 0.090 dB to 0.5941 dB at the highest percentage of 70% OPEFB filler. The dielectric properties obtained from the open-ended coaxial probe were required as input to FEM to calculate the S11 and S21 of the samples.
Polyhydroxyalkanoates (PHAs) has been paid great attention because of its useful thermoplastic properties and complete degradation in various natural environments. But, at industrial level, the successful commercialization of PHAs is limited by the high production cost due to the expensive carbon source and recovery processes. Pseudomonas mendocina PSU cultured for 72 h in mineral salts medium (MSM) containing 2% (v/v) biodiesel liquid waste (BLW) produced 79.7 wt% poly(3-hydroxybutyrate) (PHB) at 72 h. In addition, this strain produced 43.6 wt% poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) with 8.6 HV mol% at 60 h when added with 0.3% sodium propionate. The synthesized intracellular PHA granules were recovered and purified by the recently reported biological method using mealworms. The weight average molecular weight (Mw ) and number average molecular weight (Mn ) of the biologically extracted PHA were higher than that from the chloroform extraction with comparable melting temperature (Tm ) and high purity. This study has successfully established a low-cost process to synthesize PHAs from BLW and subsequently confirmed the ability of mealworms to extract PHAs from various kinds of bacterial cells.