Medium Chain Length Polyhydroxyalkanoates Research Articles

Polyenes in Medium Chain Length Polyhydroxyalkanoate (mcl-PHA) Biopolymer Microspheres with Reduced Toxicity and Improved Therapeutic Effect against Candida Infection in Zebrafish Model.

Immobilizing antifungal polyenes such as nystatin (Nys) and amphotericin B (AmB) into biodegradable formulations is advantageous compared to free drug administration providing sustained release, reduced dosing due to localized targeting and overall reduced systemic drug toxicity. In this study, we encapsulated Nys and AmB in medium chain length polyhydroxyalkanoates (mcl-PHA) microspheres (7–8 µm in diameter). The obtained formulations have been validated for antifungal activity in vitro against a panel of pathogenic fungi including species of Candida, Aspergillus, Microsporum and Trichophyton genera and toxicity and efficacy in vivo using the zebrafish model of disseminated candidiasis. While free polyenes, especially AmB, were highly toxic to zebrafish embryos at the effective (MIC) doses, after their loading into mcl-PHA microspheres, inner organ toxicity and teratogenicity associated with both drugs were not observed, even at 100 × MIC doses. The obtained mcl-PHA/polyene formulations have successfully eradicated C. albicans infection and showed an improved therapeutic profile in zebrafish by enhancing infected embryos survival. This approach is contributing to the antifungal arsenal as polyenes, although the first broad-spectrum antifungals on the market are still the gold standard for treatment of fungal infections.

Bioprospecting and Molecular Identification of Used Transformer Oil-Degrading Bacteria for Bioplastics Production

One of the major impediments to the commercialization of biodegradable plastic is the high cost of substrate. Consequently, there is a continuous search for effective microorganisms and cheaper carbon substrates to reduce the high production cost. In this study, waste transformer oil-degrading bacteria were isolated from soil, wastewater, and sediment samples, using a mineral salt medium (MSM) supplemented with 1% waste transformer oil as the sole carbon source. The isolates were screened for polyhydroxyalkanoates (PHA) production using Nile red staining and fluorescence microscopy. PHA granules accumulation was confirmed using transmission electron microscopy. Oil degradation analysis was accomplished using solvent extraction and gravimetric methods whereas, the bacteria were identified using 16S DNA sequence homology. A total of 62 transformer oil-degrading bacteria were isolated, out of which 16 (26%) showed positive results for Nile red fluorescence microscopy. The identified organisms belong to four different taxonomic genera of Acinetobacter, Bacillus, Proteus, and Serratia. The percentage of oil degradation observed among the different isolates ranged between 19.58% and 57.51%. Analysis of the PHA extracted from the selected isolate revealed the presence of medium chain length polyhydroxyalkanoates (mcl-PHA). The findings of this work have further highlighted the diversity of the bacteria capable of utilizing waste streams such as waste transformer oil. Consequently, the isolates can be explored as agents of converting waste transformer oil into bioplastics.

Microbial conversion of lignin rich biomass hydrolysates to medium chain length polyhydroxyalkanoates (mcl-PHA) using Pseudomonas putida KT2440

As world moves toward increasing number of products being produced from renewable lignocellulosic agricultural and forest residues, the major classes of products that will shift to greener routes on priority are energy, fuels, and materials in that order. In materials segment, polyhydroxyalkanoates are an emerging class of biopolyesters with several potential industrial uses. The present work investigates medium chain length polyhydroxyalkanoates (mcl-PHA) producing capabilities of Pseudomonas putida KT2440 from a mixture of compounds produced from lignocellulosic biomass deconstruction. The hydrolysates obtained from nitric acid pretreatment of lignin rich cotton stalk (CS) and palm empty fruit bunch (EFB) were used as substrates for production of mcl-PHA. Presence of 3-hydroxydecanoate and 3-hydroxyocytanoate observed on GC-MS confirmed PHA accumulation in the cells. PHA accumulation was estimated between 20% and 35% of cell dry weight when grown on both model substrates as well as biomass hydrolysates. PHA titers obtained on hydrolysates of CS and EFB were 0.24 g/L and 0.21 g/L, respectively.

Modelling Cell Growth and Polyhydroxyalkanoate (PHA) Polymer Synthesis by Pseudomonas Putida LS46 under Oxygen-Limiting Conditions

Background: Polyhydroxyalkanoates (PHAs) are biodegradable, biocompatible, and non-toxic polymers synthesized by bacteria that may be used to displace some petroleum-based plastic materials. One of the major barriers to the commercialization of PHA biosynthesis is the high cost of production. Objective: Oxygen-limitation is known to greatly influence bacterial cell growth and PHA production. In this study, the growth and synthesis of medium chain length PHAs (mcl-PHAs) by Pseudomonas putida LS46, cultured in batch-mode with octanoic acid, under oxygen-limited conditions, was modeled. Methods: Four models, including the Monod model, incorporated Leudeking-Piret (MLP), the Moser model incorporated Leudeking-Piret (Moser-LP), the Logistic model incorporated Leudeking- Piret (LLP), and the Modified Logistic model incorporated Leudeking-Piret (MLLP) were investigated. Kinetic parameters of each model were calibrated using the multi-objective optimization algorithm, Pareto Archived Dynamically Dimensioned Search (PA-DDS), by minimizing the sum of absolute error (SAE) for PHA production and growth simultaneously. Results and Conclusions: Among the four models, MLP and Moser-LP models adequately represented the experimental data for oxygen-limited conditions. However, the MLP and Moser-LP models could not adequately simulate PHA production under oxygen-excess conditions. Modeling cell growth and PHA will assist in the development of a strategy for industrial-scale production.

Biosynthesis of Medium Chain Length Polyhydroxyalkanoates (Mcl-Pha) by Activated Sludge: Impact of Phosphorus Limitation on Pha Production and Microbial Diversity

Biosynthesis of Medium Chain Length Polyhydroxyalkanoates (Mcl-Pha) by Activated Sludge: Impact of Phosphorus Limitation on Pha Production and Microbial Diversity

Valorisation of waste cooking oil using mixed culture into short- and medium-chain length polyhydroxyalkanoates: Effect of concentration, temperature and ammonium

The production of polyhydroxyalkanoates (PHAs) from waste cooking oil (WCO) by a mixed culture was investigated in the present study at increasing WCO concentrations, temperature and ammonium availability. The PHA production was done in two steps: in the first step, a mixed culture was enriched in PHA-accumulating bacteria from activated sludge in a sequencing batch reactor operated in a feast-famine mode and in the second step the PHA accumulation by the enriched mixed culture was assessed in a batch reactor. In the enrichment step, two substrates, WCO and nonanoic acid were used for enrichment and in the PHA accumulation step only WCO was used. It was not possible to enrich a mixed culture in PHA-accumulating bacteria using WCO as substrate due to the development of filamentous bacteria causing foam formation and bulking in the reactor. However, our results showed that the mixed culture continuously fed with nonanoic acid was enriched in PHA-accumulating bacteria. This enriched culture accumulated both scl- and mcl-PHA using WCO as substrate. The maximum PHA accumulation capacity of this mixed culture from WCO was 38.2% cdw. Increasing the temperature (30–40 ℃) or WCO concentrations (5–20 g/l) increased the PHA accumulation capacity of the mixed culture and the ratios of scl-PHA to mcl-PHA. The presence of ammonium increased PHA accumulation (21.9% cdw) compared to the complete absence of ammonium (5.8% cdw). The thermal characterization of the PHA exhibited the advantageous properties of both scl- and mcl-PHA, i.e., higher melting temperature (152–172 ℃) similar to scl-PHA and a lower degree of crystallinity (12%) similar to mcl-PHA. This is the first study to report the potential of open mixed culture to produce scl- and mcl-PHA from WCO and thus contributing to the understanding of sustainable polymer production.

Optimization of growth conditions for the biosynthesis of medium-chain length polyhydroxyalkanoates from Bacillus megaterium DSM 509: experimental analysis, statistical modelling, and characterization

Optimization of growth conditions for the biosynthesis of medium-chain length polyhydroxyalkanoates from Bacillus megaterium DSM 509: experimental analysis, statistical modelling, and characterization

Exploring the potential of slaughterhouse waste valorization: Development and scale-up of a new bioprocess for medium-chain length polyhydroxyalkanoates production

The progressive increase of slaughterhouse waste production requires actions for both addressing an environmental issue and creating additional value within a biorefinery concept. In this regard, some of these animal by-products exhibit a significant content of fatty acids that could be efficiently converted into bioplastics such as polyhydroxyalkanoates (PHAs) by adequately performing substrate screening with producing bacterial strains and applying affordable pretreatments. One of the main challenges also relies on the difficulty to emulsify these fat-rich substrates within culture broth and make the fatty acids accessible for the producing bacteria. In this work, the potential of two fat-rich animal by-products, grease trap waste (GTW) and tallow-based jelly (TBJ), as inexpensive carbon sources for microbial growth and PHA production was evaluated for the first time. Upon substrate screening, using different pseudomonadal strains (P. resinovorans, P. putida GPo1, P. putida KT2440) and pretreatment conditions (autoclave-based, thermally-treated or saponified substrates), the highest growth and mcl-PHA production performance was obtained for P. resinovorans, thus producing up to 47% w/w mcl-PHA simply using hygienized GTW. The novel bioprocess described in this study was successfully scaled up to 5 and 15 L, resulting in CDW concentrations of 5.9–12.8 g L−1, mcl-PHA contents of 33–62% w/w and PHA yields of 0.1–0.4 gPHA g−1fatty acids, greatly depending on the substrate dosing strategy used and depending on culture conditions. Moreover, process robustness was confirmed along Test Series by the roughly stable monomeric composition of the biopolymer produced, mainly formed by 3-hydroxyoctanoate and 3-hydroxydecanoate. The research here conducted is crucial for the cost-effectiveness of mcl-PHA production along this new slaughterhouse waste-based biorefinery concept.

A polyhydroxyalkanoates bioprocess improvement case study based on four fed‐batch feeding strategies

SummaryThe modelling and optimization of a process for the production of the medium chain length polyhydroxyalkanoate (mcl‐PHA) by the bacterium Pseudomonas putida KT2440 when fed a synthetic fatty acid mixture (SFAM) was investigated. Four novel feeding strategies were developed and tested using a constructed model and the optimum one implemented in further experiments. This strategy yielded a cell dry weight of 70.6 g l−1 in 25 h containing 38% PHA using SFAM at 5 l scale. A phosphate starvation strategy was implemented to improve PHA content, and this yielded 94.1 g l−1 in 25 h containing 56% PHA using SFAM at 5 l scale. The process was successfully operated at 20 l resulting in a cell dry weight of 91.2 g l−1 containing 65% PHA at the end of a 25‐h incubation.

Developing Bioprospecting Strategies for Bioplastics Through the Large-Scale Mining of Microbial Genomes.

The accumulation of petroleum-based plastic waste has become a major issue for the environment. A sustainable and biodegradable solution can be found in Polyhydroxyalkanoates (PHAs), a microbially produced biopolymer. An analysis of the global phylogenetic and ecological distribution of potential PHA producing bacteria and archaea was carried out by mining a global genome repository for PHA synthase (PhaC), a key enzyme involved in PHA biosynthesis. Bacteria from the phylum Actinobacteria were found to contain the PhaC Class II genotype which produces medium-chain length PHAs, a physiology until now only found within a few Pseudomonas species. Further, several PhaC genotypes were discovered within Thaumarchaeota, an archaeal phylum with poly-extremophiles and the ability to efficiently use CO2 as a carbon source, a significant ecological group which have thus far been little studied for PHA production. Bacterial and archaeal PhaC genotypes were also observed in high salinity and alkalinity conditions, as well as high-temperature geothermal ecosystems. These genome mining efforts uncovered previously unknown candidate taxa for biopolymer production, as well as microbes from environmental niches with properties that could potentially improve PHA production. This in silico study provides valuable insights into unique PHA producing candidates, supporting future bioprospecting efforts toward better targeted and relevant taxa to further enhance the diversity of exploitable PHA production systems.

Evaluation of 3D-Printing Scaffold Fabrication on Biosynthetic Medium-Chain-Length Polyhydroxyalkanoate Terpolyester as Biomaterial-Ink.

Currently, the selection of materials for tissue engineering scaffolds is still limited because some tissues require flexible and compatible materials with human cells. Medium-chain-length polyhydroxyalkanoate (MCL-PHA) synthesized in microorganisms is an interesting polymer for use in this area and has elastomeric properties compatible with the human body. MCL-PHAs are elastomers with biodegradability and cellular compatibility, making them an attractive material for fabricating soft tissue that requires high elasticity. In this research, MCL-PHA was produced by fed-batch fermentation that Pseudomonas Putida ATCC 47054 was cultured to accumulate MCL-PHA by using glycerol and sodium octanoate as carbon sources. The amounts of dry cell density, MCL-PHA product per dry cells, and MCL-PHA productivity were at 15 g/L, 27%, and 0.067 g/L/h, respectively, and the components of MCL-PHA consisting of 3-hydroxydecanoate (3HD) 64.5%, 3-hydroxyoctanoate (3HO) 32.2%, and 3-hydroxyhexanoate (3HHx) 3.3%. The biosynthesized MCL-PHA terpolyester has a relatively low melting temperature, low crystallinity, and high ductility at 52 °C, 15.7%, and 218%, respectively, and considering as elastomeric polyester. The high-resolution scaffold of MCL-PHA terpolyester biomaterial-ink (approximately 0.36 mm porous size) could be printed in a selected condition with a 3D printer, similar to the optimum pore size for cell attachment and proliferation. The rheological characteristic of this MCL-PHA biomaterial-ink exhibits shear-thinning behavior, leading to good shape fidelity. The study results yielded a condition capable of fabricating an elastomer scaffold of the MCL-PHA terpolyester, giving rise to the ideal soft tissue engineering application.

Polyhydroxyalkanoate/Antifungal Polyene Formulations with Monomeric Hydroxyalkanoic Acids for Improved Antifungal Efficiency.

Novel biodegradable and biocompatible formulations of “old” but “gold” drugs such as nystatin (Nys) and amphotericin B (AmB) were made using a biopolymer as a matrix. Medium chain length polyhydroxyalkanoates (mcl-PHA) were used to formulate both polyenes (Nys and AmB) in the form of films (~50 µm). Thermal properties and stability of the materials were not significantly altered by the incorporation of polyenes in mcl-PHA, but polyene containing materials were more hydrophobic. These formulations were tested in vitro against a panel of pathogenic fungi and for antibiofilm properties. The films containing 0.1 to 2 weight % polyenes showed good activity and sustained polyene release for up to 4 days. A PHA monomer, namely 3-hydroxydecanoic acid (C10-OH), was added to the films to achieve an enhanced synergistic effect with polyenes against fungal growth. Mcl-PHA based polyene formulations showed excellent growth inhibitory activity against both Candida yeasts (C. albicans ATCC 1023, C. albicans SC5314 (ATCC MYA-2876), C. parapsilosis ATCC 22019) and filamentous fungi (Aspergillus fumigatus ATCC 13073; Trichophyton mentagrophytes ATCC 9533, Microsporum gypseum ATCC 24102). All antifungal PHA film preparations prevented the formation of a C. albicans biofilm, while they were not efficient in eradication of mature biofilms, rendering them suitable for the transdermal application or as coatings of implants.

Comprehensive optimization of tropical biomass hydrolysis for nitrogen-limited medium-chain polyhydroxyalkanoate synthesis.

Expanding the use of tropical biomass wastes for nitrogen-limited fermentation was investigated, specifically, the production of medium chain length polyhydroxyalkanoates. Comprehensive central composite design was conducted to assess pH, temperature, biomass solid loading, cellulase loading and amylase loading and their impact on the hydrolysis of palm, coconut and cassava wastes. Glucose yields of 33.3, 31.7 and 79.0% wt. with respect to total glucose were found for palm, coconut and cassava, respectively. Importantly, the impact on the total nitrogen derived during enzymatic hydrolysis of these tropical biomass was described for the first time. The level of nitrogen needs to be properly controlled as high nitrogen would result in low carbon to nitrogen ratio leading to low polyhydroxyalkanoates accumulation, but low nitrogen would hinder growth of the biopolymer producer. Maximum hydrolysate nitrogen, were 1.80, 1.55 and 0.871g/l for palm, coconut and cassava, respectively. Using the surface responses, biomass media designed for high carbon-to-nitrogen were produced and validated using Pseudomonas putida. Low glucose-carbon to nitrogen were found for palm and coconut after scale-up, leading to the majority of their polyhydroxyalkanoates not being biomass-derived. However, cassava-derived biopolymers were successfully accumulated at 9.01 and 7.13% wt. for total medium chain length polyhydroxyalkanoates and 10-carbon polyhydroxyalkanoates, respectively. This study provides an important foundation for the expansion of tropical biomass wastes for biopolymer production and other nitrogen-limited applications in general.

Towards bio-upcycling of polyethylene terephthalate

Over 359 million tons of plastics were produced worldwide in 2018, with significant growth expected in the near future, resulting in the global challenge of end-of-life management. The recent identification of enzymes that degrade plastics previously considered non-biodegradable opens up opportunities to steer the plastic recycling industry into the realm of biotechnology.Here, the sequential conversion of post-consumer polyethylene terephthalate (PET) into two types of bioplastics is presented: a medium chain-length polyhydroxyalkanoate (PHA) and a novel bio-based poly(amide urethane) (bio-PU). PET films are hydrolyzed by a thermostable polyester hydrolase yielding highly pure terephthalate and ethylene glycol. The obtained hydrolysate is used directly as a feedstock for a terephthalate-degrading Pseudomonas umsongensis GO16, also evolved to efficiently metabolize ethylene glycol, to produce PHA. The strain is further modified to secrete hydroxyalkanoyloxy-alkanoates (HAAs), which are used as monomers for the chemo-catalytic synthesis of bio-PU. In short, a novel value-chain for PET upcycling is shown that circumvents the costly purification of PET monomers, adding technological flexibility to the global challenge of end-of-life management of plastics.

Oxygen Plasma Treated-Electrospun Polyhydroxyalkanoate Scaffolds for Hydrophilicity Improvement and Cell Adhesion.

Poly(hydroxyalkanoates) (PHAs) with differing material properties, namely, the homopolymer poly(3-hydroxybutyrate), P(3HB), the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate), P(3HB-co-3HV), with a 3HV content of 25 wt.% and a medium chain length PHA, and mcl-PHA, mainly composed of 3-hydroxydecanoate, were studied as scaffolding material for cell culture. P(3HB) and P(3HB-co-3HV) were individually spun into fibers, as well as blends of the mcl-PHA with each of the scl-PHAs. An overall biopolymer concentration of 4 wt.% was used to prepare the electrospinning solutions, using chloroform as the solvent. A stable electrospinning process and good quality fibers were obtained for a solution flow rate of 0.5 mL h−1, a needle tip collector distance of 20 cm and a voltage of 12 kV for P(3HB) and P(3HB-co-3HV) solutions, while for the mcl-PHA the distance was increased to 25 cm and the voltage to 15 kV. The scaffolds’ hydrophilicity was significantly increased under exposure to oxygen plasma as a surface treatment. Complete wetting was obtained for the oxygen plasma treated scaffolds and the water uptake degree increased in all treated scaffolds. The biopolymers crystallinity was not affected by the electrospinning process, while their treatment with oxygen plasma decreased their crystalline fraction. Human dermal fibroblasts were able to adhere and proliferate within the electrospun PHA-based scaffolds. The P(3HB-co-3HV): mcl-PHA oxygen plasma treated scaffold highlighted the most promising results with a cell adhesion rate of 40 ± 8%, compared to 14 ± 4% for the commercial oxygen plasma treated polystyrene scaffold AlvetexTM. Scaffolds based on P(3HB-co-3HV): mcl-PHA blends produced by electrospinning and submitted to oxygen plasma exposure are therefore promising biomaterials for the development of scaffolds for tissue engineering.

Microbial Polyhydroxyalkanoates Granules: An Approach Targeting Biopolymer for Medical Applications and Developing Bone Scaffolds.

Microbial polyhydroxyalkanoates (PHA) are proteinaceous storage granules ranging from 100 nm to 500 nm. Bacillus sp. serve as unique bioplastic sources of short-chain length and medium-chain length PHA showcasing properties such as biodegradability, thermostability, and appreciable mechanical strength. The PHA can be enhanced by adding functional groups to make it a more industrially useful biomaterial. PHA blends with hydroxyapatite to form nanocomposites with desirable features of compressibility. The reinforced matrices result in nanocomposites that possess significantly improved mechanical and thermal properties both in solid and melt states along with enhanced gas barrier properties compared to conventional filler composites. These superior qualities extend the polymeric composites’ applications to aggressive environments where the neat polymers are likely to fail. This nanocomposite can be used in different industries as nanofillers, drug carriers for packaging essential hormones and microcapsules, etc. For fabricating a bone scaffold, electrospun nanofibrils made from biocomposite of hydroxyapatite and polyhydroxy butyrate, a form of PHA, can be incorporated with the targeted tissue. The other methods for making a polymer scaffold, includes gas foaming, lyophilization, sol–gel, and solvent casting method. In this review, PHA as a sustainable eco-friendly NextGen biomaterial from bacterial sources especially Bacillus cereus, and its application for fabricating bone scaffold using different strategies for bone regeneration have been discussed.

Bioresorbable and Mechanically Optimized Nerve Guidance Conduit Based on a Naturally Derived Medium Chain Length Polyhydroxyalkanoate and Poly(ε-Caprolactone) Blend.

Severe peripheral nerve injuries represent a large clinical problem with relevant challenges such as the development of successful synthetic scaffolds as substitutes to autologous nerve grafting. Numerous studies have reported the use of polyesters and type I collagen-based nerve guidance conduits (NGCs) to promote nerve regeneration through critical nerve defects while providing protection from external factors. However, none of the commercially available hollow bioresorbable NGCs have demonstrated superior clinical outcomes to an autologous nerve graft. Hence, new materials and NGC geometries have been explored in the literature to mimic the native nerve properties and architecture. Here, we report a novel blend of a natural medium chain length polyhydroxyalkanoate (MCL-PHA) with a synthetic aliphatic polyester, poly(ε-caprolactone) (PCL), suitable for extrusion-based high-throughput manufacturing. The blend was designed to combine the excellent ability of PHAs to support the growth and proliferation of mammalian cells with the good processability of PCL. The material exhibited excellent neuroregenerative properties and a good bioresorption rate, while the extruded porous tubes exhibited similar mechanical properties to the rat sciatic nerve. The NGCs were implanted to treat a 10 mm long sciatic nerve defect in rats, where significant differences were found between thin and thick wall thickness implants, and both electrophysiological and histological data, as well as the number of recovered animals, provided superior outcomes than the well-referenced synthetic Neurolac NGC.

The Thermal and Mechanical Properties of Medium Chain-Length Polyhydroxyalkanoates Produced by Pseudomonas putida LS46 on Various Substrates.

Medium chain-length polyhydroxyalkanoates (mcl-PHA) were produced by Pseudomonas putida LS46 cultured with a variety of carbohydrate and fatty acid substrates. The monomer compositions and molecular weights of the polymers varied greatly and was dependent on whether the substrate was metabolized via the fatty acid degradation or the de novo fatty acid synthesis pathways. The highest molecular weights were obtained from medium chain-length fatty acids, whereas low molecular weights were obtained from longer chain-length and more unsaturated fatty acids or carbohydrates. The differences in monomer compositions and molecular weights due to the choice of substrate did not affect the polymer thermal degradation point. The glass transition temperatures varied from −39.4°C to −52.7°C. The melting points, when observed, ranged from 43.2°C to 51.2°C. However, a profound substrate effect was observed on the crystallinity of these polymers. Reduced crystallinity was observed when the monomer compositions deviated away from C8–C10 monomer lengths. The highest crystallinity was observed from medium chain-length fatty acids, which resulted in polymers with the highest tensile strength. The polymer produced from octanoic acid exhibited the highest tensile strength of 4.3 MPa with an elongation-at-break of 162%, whereas the polymers produced from unsaturated, long-chain fatty acids remained amorphous. A comparative analysis of the substrate effect on the physical-mechanical and thermal properties of mcl-PHAs better clarifies the relationship between the monomer composition and their potential applications, and also aids to direct future PHA synthesis research toward properties of interest.

Productivity of Pseudomonas putida TISTR 1522 in polyhydroxyalkanoates (PHAs) production from saponified palm oil.

Polyhydroxyalkanoates (PHAs) have attracted attention as an environmentally degradable bioplastic which potentially replaces synthetic polymers used in a wide range of industries. One of most promising microorganisms for the production of PHAs is Pseudomonas putida. In this study, we purpose to develop sustainable processes to convert abundant palm oil available in local market to high value PHAs and optimize PHAs production by Pseudomonas putida TISTR 1522 from saponified palm oil. We found that the highest yield of PHAs production (0.95 g/L, 40.15%) was obtained in culture medium supplemented with 1% (w/v) fatty acid salt by P. putida TISTR 1522 after 24-h cultivation. The intracellular PHAs were located in granules inside the cells, which fluoresced bright yellow by staining with Nile red. The physical appearance of intracellular PHAs investigated by transmission electron microscope (TEM) revealed that PHAs accumulate in granules, about 3-10 granules per cell. These granules are white and roundish-shaped with 0.3-0.5-μm diameter. The 1H NMR spectrum represented the typical characters of medium-chain length-PHAs. This variation of all parameters was successfully demonstrated a good intracellular PHAs accumulation in P. putida TISTR 1522 by fatty acid salt utilization.

Enhanced medium chain length-polyhydroxyalkanoate production by co-fermentation of lignin and holocellulose hydrolysates

Biological lignin and holocellulose conversion to medium chain length-polyhydroxyalkanoates (mcl-PHA) as an excellent platform for bioplastics production and attractive alternative for current biorefinery schemes.