Type Of Polyhydroxyalkanoates Research Articles

Coastal and deep-sea biodegradation of polyhydroxyalkanoate microbeads

Microbeads find widespread usage in personal care items and cosmetics, serving as exfoliants or scrubbing agents. Their micro-scale size poses challenges in effective drainage capture and given their origin from non-biodegradable oil-based plastics, this contributes substantially to marine pollution. In this study, microbeads were prepared by a simple yet scalable melt homogenization method using four types of polyhydroxyalkanoates (PHA), namely poly[(R)-3-hydroxybutyrate] (P(3HB)), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyvalerate] (P(3HB-co-3HV)), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (P(3HB-co-3HHx)) and poly[(R)-3-hydroxybutyrate-co-(R)-4-hydroxyvalerate] (P(3HB-co-4HB)). Microbeads with different surface smoothness, compressive strength (6.2–13.3 MPa) and diameter (from 1 ~ 150 μm) could be produced. The microbeads were subjected to a comprehensive degradation analysis using three techniques: enzymatic, Biochemical Oxygen Demand (BOD) evaluations, and in situ degradation tests in the deep-sea off Misaki Port in the northern Pacific Ocean (depth of 757 m). Qualitatively, results from enzymatic and in situ degradation demonstrated significant degradation within one week and five months, respectively. Quantitatively, BOD findings indicated that all PHA microbeads degraded similarly to cellulose (~ 85% biodegradability in 25 days). In conclusion, PHA microbeads from this study exhibit promising potential as alternatives to conventional non-biodegradable microbeads.

Unveiling the repressive mechanism of a PPS-like regulator (PspR) in polyhydroxyalkanoates biosynthesis network.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a type of polyhydroxyalkanoates (PHA) that exhibits numerous outstanding properties and is naturally synthesized and elaborately regulated in various microorganisms. However, the regulatory mechanism involving the specific regulator PhaR in Haloferax mediterranei, a major PHBV production model among Haloarchaea, is not well understood. In our previous study, we showed that deletion of the phosphoenolpyruvate (PEP) synthetase-like (pps-like) gene activates the cryptic phaC genes in H. mediterranei, resulting in enhanced PHBV accumulation. In this study, we demonstrated the specific function of the PPS-like protein as a negative regulator of phaR gene expression and PHBV synthesis. Chromatin immunoprecipitation (ChIP), in situ fluorescence reporting system, and in vitro electrophoretic mobility shift assay (EMSA) showed that the PPS-like protein can bind to the promoter region of phaRP. Computational modeling revealed a high structural similarity between the rifampin phosphotransferase (RPH) protein and the PPS-like protein, which has a conserved ATP-binding domain, a His domain, and a predicted DNA-binding domain. Key residues within this unique DNA-binding domain were subsequently validated through point mutation and functional evaluations. Based on these findings, we concluded that PPS-like protein, which we now renamed as PspR, has evolved into a repressor capable of regulating the key regulator PhaR, and thereby modulating PHBV synthesis. This regulatory network (PspR-PhaR) for PHA biosynthesis is likely widespread among haloarchaea, providing a novel approach to manipulate haloarchaea as a production platform for high-yielding PHA. KEY POINTS: • The repressive mechanism of a novel inhibitor PspR in the PHBV biosynthesis was demonstrated • PspR is widespread among the PHA accumulating haloarchaea • It is the first report of functional conversion from an enzyme to a trans-acting regulator in haloarchaea.

Biorefinery of volatile fatty acids for the synthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) using Paracoccus sp. TOH: Fermentation performance and metabolic pathway

The residual microbes and uncertain volatile fatty acids (VFAs) components in anaerobic fermentation hydrolysate cause microbial contamination and multi-type polyhydroxyalkanoates (PHA) generation in pure microbial synthesis systems. Meanwhile, the regulation of biosynthesis of poly(3-hydroxybuturate-co-3-hydroxyvalerate) (PHBV), one type of excellent PHA, by re-utilization of waste-organics as carbon substrates is still elusive. In this study, four pretreatment methods for sludge hydrolysate containing 4746 mg-COD·L−1 of VFAs were established to assess the accumulation of PHBV by using Paracoccus sp. TOH as the chassis. Results showed that the autoclaving system exhibited an optimal PHBV concentration of 202.9 mg L−1 with the highest relative abundance of Paracoccus sp. TOH (92.4%). Valeric acid was identified as the primary substrate for 3-hydroxyvalerate accumulation, and the highest PHBV production of 3.76 g L−1 was achieved using 3 g L−1 valeric acid and 5 g L−1 glycerol as co-substrates. The increased carbon metabolism flows through the sequential action of acyl-coenzyme A dehydrogenase and enoyl-coenzyme A hydratase in the fatty acid β-oxidation pathway facilitated PHBV accumulation. This study provides a blueprint for the microbial synthesis of PHBV with oriented acid production from organic wastes.

Nature-inspired material binding peptides with versatile polyester affinities and binding strengths

Biodegradable polyesters, such as polyhydroxyalkanoates (PHAs), are having a tremendous impact on biomedicine. However, these polymers lack functional moieties to impart functions like targeted delivery of molecules. Inspired by native GAPs, such as phasins and their polymer-binding and surfactant properties, we generated small material binding peptides (MBPs) for polyester surface functionalization using a rational approach based on amphiphilicity. Here, two peptides of 48 amino acids derived from phasins PhaF and PhaI from Pseudomonas putida, MinP and the novel-designed MinI, were assessed for their binding towards two types of PHAs, PHB and PHOH. In vivo, fluorescence studies revealed selective binding towards PHOH, whilst in vitro binding experiments using the Langmuir-Blodgett technique coupled to ellipsometry showed KD in the range of nM for all polymers and MBPs. Marked morphological changes of the polymer surface upon peptide adsorption were shown by BAM and AFM for PHOH. Moreover, both MBPs were successfully used to immobilize cargo proteins on the polymer surfaces. Altogether, this work shows that by redesigning the amphiphilicity of phasins, a high affinity but lower specificity to polyesters can be achieved in vitro. Furthermore, the MBPs demonstrated binding to PET, showing potential to bind cargo molecules also to synthetic polyesters.

A review on poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] and genetic modifications that affect its production.

Polyhydroxyalkanoates (PHAs) have garnered global attention to replace petroleum-based plastics in certain applications due to their biodegradability and sustainability. Among the different types of PHAs, poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] copolymer has similar properties to commodity plastics, making them a suitable candidate to replace certain types of single-use plastics, medical devices, and packaging materials. The degradation rate of P(3HB-co-3HHx) is faster than the commercial petroleum-based plastics which take a very long time to be degraded, causing harmful pollution to both land and marine ecosystem. The biodegradability of the P(3HB-co-3HHx) is also dependent on its 3HHx molar composition which in turn influences the crystallinity of the material. Various metabolic pathways like the common PHA biosynthesis pathway, which involves phaA, phaB, and phaC, β-oxidation, and fatty acids de novo synthesis are used by bacteria to produce PHA from different carbon sources like fatty acids and sugars, respectively. There are various factors affecting the 3HHx molar composition of P(3HB-co-3HHx), like PhaCs, the engineering of PhaCs, and the metabolic engineering of strains. It is crucial to control the 3HHx molar composition in the P(3HB-co-3HHx) as it will affect its properties and applications in different fields.

Co-production of polyhydroxyalkanoates and levan by Halomonas smyrnensis AAD6T

The simultaneous production of microbial polymers levan and poly[3-hydroxybutyrate] (PHB), a type of polyhydroxyalkanoates, was investigated in this work. The study involved the fermentation of sucrose and molasses by H. smyrnensis AAD6T (BAE2 strain) to produce PHB (intracellular) and levan (extracellular). Both polymers were isolated and characterized by FTIR. Levan was also characterized by thin-layer chromatography (TLC) and viscosimetric analysis. The amount of biomass was 25 g until the end of fermentation. The PHB rate was 0.015 g in both media and the average PHB productivity was 6.0 x 10-4 g PHB/g biomass. The highest rate of levan was 9 g/L in the range of 72–80 h, in the molasses-based medium. The FTIR spectra showed specific signals for each of the polymers, such as the peak at 1700 for the carbonyl group of esters for the PHB and signals at 900 and 800, which are typical signals for levan fructose rings. Furthermore, acid hydrolysis of levan revealed that it was formed only by fructose, as confirmed by TLC With this study, H. smyrnensis AAD6T BAE2 co-produced PHB and levan using a low-cost carbon source, showing great potential in reducing biopolymer manufacturing costs.

Characterization of the polyhydroxyalkanoate (PHA) synthase from Ideonella sakaiensis, a bacterium that is capable of degrading and assimilating poly(ethylene terephthalate)

• Characterization of PhaC from Ideonella sakaiensis 201-F6, a bacterium that is capable of degrading and assimilating PET. • PhaC Is polymerizes five types of monomers: 3HB, 3HV, 4HB, 5HV and 3H4MV. • Provides insights into the substrate specificity of PhaC Is , which is important for the metabolic engineering of I. sakaiensis that produces various types of PHA from PET. Ideonella sakaiensis 201-F6 is a bacterium capable of degrading and assimilating poly(ethylene terephthalate) (PET). A recent study revealed its capability in accumulating intracellular polyhydroxyalkanoate (PHA) at high levels using PET as a carbon source. The finding demonstrated that I. sakaiensis can mediate the direct conversion of non-biodegradable PET into environment-friendly PHA, providing a new approach for PET recycling. The PHA synthase gene of I. sakaiensis ( phaC Is ) was characterized in this study. Phylogenetic analysis of the PhaC protein sequences indicated PhaC Is belong to class I PhaC. To carry out in vivo characterization of PhaC Is , phaC Is was cloned and heterologously expressed in a PHA-negative mutant strain Cupriavidus necator PHB⁻4. The heterologous expression of the PhaC Is in PHB⁻4/pBBR1- C Is resulted in poly(3-hydroxybutyrate) [P(3HB)] accumulation of up to 71 wt% of the cell dry weight. In addition, the PhaC Is was shown to be able to polymerize 5 types of monomers, 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 4-hydroxybutyrate (4HB), 5-hydroxyvalerate (5HV), and 3-hydroxy-4-methylvalerate (3H4MV). In vitro enzymatic assay showed that PhaC Is was capable of catalyzing ( R )-3HB-CoA with a specific activity of 11.6 ± 0.6 nmol/min/mg and displayed an initial lag-phase. This study provides a basic information of PhaC Is that allows us to create variants applied for the metabolic engineering of I. sakaiensis that produces various types of PHA from PET.

Potential of mealworms used in polyhydroxyalkanoate/bioplastic recovery as red hybrid tilapia (Oreochromis sp.) feed ingredient

Polyhydroxyalkanoates (PHAs) are bio-based polymers produced in bacterial cells to replace some petrochemical plastics. It has always been a challenge to commercialise PHA due in part to the costly recovery processes of the PHA granules from the bacterial cells. The biological approach of using mealworms, Tenebrio molitor, for the recovery of PHA from the bacterial cells is a newly established method that is at the scale-up stage. On the other hand, the aquaculture feed industry needs a low-cost mealworm meal as a protein source. We aimed at studying the nutritional value of the mealworms (which are by-products) used for the poly(3-hydroxybutyrate) (PHB) (the most common type of PHA) recovery from the bacterial and examining the effect of the mealworms on the growth performance, and feed utilization efficiency of red hybrid tilapia (Oreochromis sp.). The cells were fed to the mealworms to digest the proteinaceous cellular materials and excrete the PHB granules in the form of fecal pellets. The resulting mealworms were used as fishmeal replacement to formulate five isonitrogenous (35% crude protein) and isolipidic (8% lipid) diets at mealworm meal (MwM) inclusion levels of 0% (MwM0/control diet), 25% (MwM25), 50% (MwM50), 75% (MwM75) or 100% (MwM100). The results showed good nutritive value mealworms [high protein (75%), low-lipid (10%)] and up to 75% MwM inclusion diet was good in supplying satisfactory nutrients and energy to the red hybrid tilapia. This approach is beneficial in a way that minimal cost was involved in recovering kilograms of PHB and the proteins, lipids, and minerals from the bacterial cells do not end up as wastes but in turn, are used as nutrition by the larvae.

A New Wave of Industrialization of PHA Biopolyesters.

The ever-increasing use of plastics, their fossil origin, and especially their persistence in nature have started a wave of new innovations in materials that are renewable, offer the functionalities of plastics, and are biodegradable. One such class of biopolymers, polyhydroxyalkanoates (PHAs), are biosynthesized by numerous microorganisms through the conversion of carbon-rich renewable resources. PHA homo- and heteropolyesters are intracellular products of secondary microbial metabolism. When isolated from microbial biomass, PHA biopolymers mimic the functionalities of many of the top-selling plastics of petrochemical origin, but biodegrade in soil, freshwater, and marine environments, and are both industrial- and home-compostable. Only a handful of PHA biopolymers have been studied in-depth, and five of these reliably match the desired material properties of established fossil plastics. Realizing the positive attributes of PHA biopolymers, several established chemical companies and numerous start-ups, brand owners, and converters have begun to produce and use PHA in a variety of industrial and consumer applications, in what can be described as the emergence of the “PHA industry”. While this positive industrial and commercial relevance of PHA can hardly be described as the first wave in its commercial development, it is nonetheless a very serious one with over 25 companies and start-ups and 30+ brand owners announcing partnerships in PHA production and use. The combined product portfolio of the producing companies is restricted to five types of PHA, namely poly(3-hydroxybutyrate), poly(4-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxybutyrate-co-4-hydroxybutyrate), and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate), even though PHAs as a class of polymers offer the potential to generate almost limitless combinations of polymers beneficial to humankind. To date, by varying the co-monomer type and content in these PHA biopolymers, their properties emulate those of the seven top-selling fossil plastics, representing 230 million t of annual plastics production. Capacity expansions of 1.5 million t over the next 5 years have been announced. Policymakers worldwide have taken notice and are encouraging industry to adopt biodegradable and compostable material solutions. This wave of commercialization of PHAs in single-use and in durable applications holds the potential to make the decisive quantum leap in reducing plastic pollution, the depletion of fossil resources, and the emission of greenhouse gases and thus fighting climate change. This review presents setbacks and success stories of the past 40 years and the current commercialization wave of PHA biopolymers, their properties, and their fields of application.

Enhancing the biological reductive dechlorination of trichloroethylene with PHA from mixed microbial cultures (MMC)

The biological reductive dechlorination (BRD) process is often limited by the lack of suitable electron donors, requiring the supply of long-lasting substrates to enhance the biological metabolism. Among the others, polyhydroxyalkanoates (PHA) are a particularly interesting slow-release source of electron-donors, being entirely biodegradable polyesters. Generally, industrial PHA production processes are based on pure culture fermentation with high related costs. In recent years, innovative and cost-effective PHA production from mixed microbial cultures (MMC) and waste feedstocks is attracting considerable attention. This research, for the first time, investigated the effect of distinctive types of PHA on the BRD process of trichloroethylene (TCE), in a continuous-flow lab-scale system using two PHA materials with different purity grade produced from MMC at pilot scale in comparison with a commercial PHA produced from pure culture. Promising results have been obtained with non-extracted MMC-PHA, a material consisting of both PHA (56%, w/w) and microbial cells, with constant production of acids over 110 days, which stimulated a nearly complete TCE dechlorination with the non-toxic ethene as the main byproduct (approximately 92%). Dehalococcoides mccartyi was the main microorganism responsible for TCE dechlorination process, representing ≥ 54.05% of the bacterial population, mainly carrying the reductive dehalogenase genes tceA and vcrA (≥9.17E+08 and ≥2.01E+07 gene copies/g of PHA or sand, respectively). This finding suggests the possibility to directly use PHA-rich biomass deriving from MMC production process, which does not require any polluting and expensive extraction procedures, as a novel material in the field of groundwater remediation with noticeable economic and environmental advantages.

On-Line Control of Feast/Famine Cycles to Improve PHB Accumulation during Cultivation of Mixed Microbial Cultures in Sequential Batch Reactors.

Production of polyhydroxyalkanoates (PHA) has generated great interest as building blocks for bioplastic production. Their production using mixed microbial cultures represents an interesting alternative, since it enables the use of organic wastes as a carbon source. Feast/famine strategy is a common way to promote selection of microorganisms with PHA accumulation capacity. However, when using waste sources, changes in substrate concentration are expected, that may affect performance and efficiency of the process. This study showed how the dissolved oxygen level can be used for online control of the cycle time, ensuring that the desired feast/famine ratio is effectively applied. An operation strategy is presented and validated, using sequential batch reactors fed with acetate as the carbon source. Production of polyhydroxybutyrate (PHB) was studied, which is the expected type of PHA to be synthetized when using acetate as substrate. Two reactors were operated by applying the proposed control strategy, to provide F/F ratios of 0.2 and 0.6, respectively. A third reactor was operated with a fixed cycle time, for comparison purposes. Results showed that the reactor that operated at an F/F ratio of 0.6 promoted higher biomass productivity and PHB content, as a result of a better use of available time, preventing unnecessary long famine times. The application of the tested strategy is a simple a reliable way to promote a better performance of feast/famine-based bioreactors involving mixed microbial cultures for PHB production.

Current Advances towards 4-Hydroxybutyrate Containing Polyhydroxyalkanoates Production for Biomedical Applications.

Polyhydroxyalkanoates (PHA) are polyesters having high promise in biomedical applications. Among different types of PHA, poly-4-hydroxybutyrate (P4HB) is the only polymer that has received FDA approval for medical applications. However, most PHA producing microorganisms lack the ability to synthesize P4HB or PHA comprising 4-hydroxybutyrate (4HB) monomer due to their absence of a 4HB monomer supplying pathway. Thus, most microorganisms require supplementation of 4HB precursors to synthesize 4HB polymers. However, usage of 4HB precursors incurs additional production cost. Therefore, researchers have adopted strategies to reduce the cost, such as utilizing low-cost substrate as well as constructing 4HB monomer supplying pathways in microorganisms. We herein summarize the biomedical applications of P4HB, the natural producers of 4HB polymer, and the various strategies that have been applied in producing 4HB polymers in non-4HB producing microorganisms. It is expected that the readers would gain a vivid idea on the different strategic developments in the field of 4HB polymer production.

The Synthesis, Characterization and Applications of Polyhydroxyalkanoates (PHAs) and PHA-Based Nanoparticles.

Polyhydroxyalkanoates (PHAs) are storage granules found in bacteria that are essentially hydroxy fatty acid polyesters. PHA molecules appear in variety of structures, and amongst all types of PHAs, polyhydroxybutyrate (PHB) is used in versatile fields as it is a biodegradable, biocompatible, and ecologically safe thermoplastic. The unique physicochemical characteristics of these PHAs have made them applicable in nanotechnology, tissue engineering, and other biomedical applications. In this review, the optimization, extraction, and characterization of PHAs are described. Their production and application in nanotechnology are also portrayed in this review, and the precise and various production methods of PHA-based nanoparticles, such as emulsion solvent diffusion, nanoprecipitation, and dialysis are discussed. The characterization techniques such as UV-Vis, FTIR, SEM, Zeta Potential, and XRD are also elaborated.

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

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

Recyclable Polyhydroxyalkanoates via a Regioselective Ring-Opening Polymerization of α,β-Disubstituted β-Lactone Monomers

The economical and societal issues associated with the depletion of fossil feedstocks and severe accumulation of nondegradable plastics in nature have pushed the increasing efforts toward the development of biodegradable polymers with unique recyclability. The bacterial polyhydroxyalkanoates (PHAs) represent a unique class of biodegradable polymers that have found a variety of applications in packaging, agricultural, biomedical, and pharmaceutical fields. Due to the high production costs and limited volumes for the bacterial fermentation process, the chemical synthesis of PHAs via ring-opening polymerization of lactone monomers has attracted intensive attention. However, most β-lactones feature a substituent group at the β-position, and they cannot be recycled back into the monomers or other intermediates in quantitative yields. Rare examples were reported regarding the synthesis of PHAs based on β-lactones with α,β-disubstituted groups or a ring fusion at both α- and β-positions. Here, we report a type of β-lactone fused with a five-membered ring fusion at α- and β-positions (cis-6-oxabicyclo[3.2.0]heptan-7-one, β-CPL), from the ring-expansive carbonylation of cyclopentene oxide (CPO). With the efficient catalysts, the ring-opening polymerization of β-CPL under ambient conditions produces a novel type of polyhydroxyalkanoates with controlled linear and/or cyclic topologies; the produced PHAs feature a high melting temperature (∼185 °C) and high molecular weight (Mn = 1.01 × 105 g mol–1, Đ = 1.36) via a regioselective fashion through an O-acyl cleavage. In particular, the quantitative formation of α,β-unsaturated carboxylic acid during pyrolysis represents a rare example that the recyclability can be accomplished via a transformation of α,β-unsaturated carboxylic acid to β-lactone monomers.

Antibacterial Composite Materials Based on the Combination of Polyhydroxyalkanoates With Selenium and Strontium Co-substituted Hydroxyapatite for Bone Regeneration.

Due to the threat posed by the rapid growth in the resistance of microbial species to antibiotics, there is an urgent need to develop novel materials for biomedical applications capable of providing antibacterial properties without the use of such drugs. Bone healing represents one of the applications with the highest risk of postoperative infections, with potential serious complications in case of bacterial contaminations. Therefore, tissue engineering approaches aiming at the regeneration of bone tissue should be based on the use of materials possessing antibacterial properties alongside with biological and functional characteristics. In this study, we investigated the combination of polyhydroxyalkanoates (PHAs) with a novel antimicrobial hydroxyapatite (HA) containing selenium and strontium. Strontium was chosen for its well-known osteoinductive properties, while selenium is an emerging element investigated for its multi-functional activity as an antimicrobial and anticancer agent. Successful incorporation of such ions in the HA structure was obtained. Antibacterial activity against Staphylococcus aureus 6538P and Escherichia coli 8739 was confirmed for co-substituted HA in the powder form. Polymer-matrix composites based on two types of PHAs, P(3HB) and P(3HO-co-3HD-co-3HDD), were prepared by the incorporation of the developed antibacterial HA. An in-depth characterization of the composite materials was conducted to evaluate the effect of the filler on the physicochemical, thermal, and mechanical properties of the films. In vitro antibacterial testing showed that the composite samples induce a high reduction of the number of S. aureus 6538P and E. coli 8739 bacterial cells cultured on the surface of the materials. The films are also capable of releasing active ions which inhibited the growth of both Gram-positive and Gram-negative bacteria.

Production of Biohydrogen and/or Poly-β-hydroxybutyrate by Rhodopseudomonas sp. Using Various Carbon Sources as Substrate.

The polyhydroxyalkanoates (PHA) are family of biopolyesters synthesized by numerous bacteria which are attracting a great attention due to their thermoplastic properties. Polyhydroxybutyrate (PHB) is the most common type of PHA which presents thermoplastic and biodegradable properties. It is synthesized under stressful conditions by heterotrophic bacteria and many photosynthetic microorganisms such as purple non-sulfur bacteria and cyanobacteria. Biological hydrogen (H2) production is being evaluated for use as a fuel since it is a promising substitute for carbonaceous fuels owing to its high conversion efficiency and high specific content. In the present work, the purple non-sulfur photosynthetic bacterium Rhodopseudomonas sp. for the simultaneous H2 photo-evolution and poly-β-hydroxybutyrate (PHB) production has been investigated. Three different types of carbon sources were tested in the presence of glutamate as a nitrogen source in a batch cultivation system, under continuous irradiance. The results indicated the fact that the type of carbon source in the culture broth affects in various ways the metabolic activity of the bacterial biomass, as evidenced by the production of PHB and/or H2 and biomass. The best carbon source for PHB accumulation and H2 production by Rhodopseudomonas sp. turned out to be the acetate, having the highest H2 production (2286 mL/L) and PHB accumulation (68.99 mg/L, 18.28% of cell dry weight).

Established and advanced approaches for recovery of microbial polyhydroxyalkanoate (PHA) biopolyesters from surrounding microbial biomass

Abstract Downstream processing for recovery of microbial polyhydroxyalkanoate (PHA) biopolyesters from biomass constitutes an integral part of the entire PHA production chain; beside the feedstocks used for cultivation of PHA-production strains, this process is currently considered the major cost factor for PHA production. Besides economic aspects, PHA recovery techniques need to be sustainable by avoiding excessive use of (often precarious!) solvents, other hazardous chemicals, non-recyclable compounds, and energy. Moreover, the applied PHA recovery method is decisive for the molecular mass and purity of the obtained product, and the achievable recovery yield. In addition to the applied method, also the PHA content in biomass is decisive for the feasibility of a selected technique. Further, not all investigated recovery techniques are applicable for all types of PHA (crystalline versus amorphous PHA) and all PHA-producing microorganisms (robust versus fragile cell structures). The present review shines a light on benefits and shortcomings of established solvent-based, chemical, enzymatic, and mechanical methods for PHA recovery. Focus is dedicated on innovative, novel recovery strategies, encompassing the use of “green” solvents, application of classical “PHA anti-solvents” under pressurized conditions, ionic liquids, supercritical solvents, hypotonic cell disintegration for release of PHA granules, switchable anionic surfactants, and even digestion of non-PHA biomass by animals. The different established and novel techniques are compared in terms of PHA recovery yield, product purity, impact on PHA molar mass, scalability to industrial plants, and demand for chemicals, energy, and time.

Evaluation of BP-M-CPF4 polyhydroxyalkanoate (PHA) synthase on the production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from plant oil using Cupriavidus necator transformants

Among the various types of polyhydroxyalkanoate (PHA), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)] has a high potential to serve as commercial bioplastic due to its striking resemblance to petroleum-based plastics. In this study, five different genotypes of Cupriavidusnecator transformants harbouring the phaCBP-M-CPF4 gene (including PHB¯4/pBBR1-CBP-M-CPF4) were developed to evaluate the efficiency of 3HHx monomer incorporation. The fraction of 3-hydroxyhexanoate (3HHx) monomer that was incorporated into the PHA synthesized by these C. necator transformants using palm oil as the sole carbon source, was examined. Overall, co-expression of enoyl-CoA hydratase gene (phaJ1) from Pseudomonas aeruginosa, along with PHA synthase (PhaC), increased the 3HHx composition in the PHA copolymer. The differences in the enzyme activities of β-ketothiolase (PhaACn) and NADPH-dependent acetoacetyl-CoA reductase (PhaBCn) of the C. necator mutant hosts used in this study, were observed to alter the 3HHx composition and molecular weight of the PHA copolymer produced. The 3HHx fractions in the P(3HB-co-3HHx) produced by these C. necator transformants ranged between 1 and 18 mol%, while the weight-average molecular weight ranged from 0.7 × 106 to 1.8 × 106 Da. PhaCBP-M-CPF4 displayed a typical initial lag-phase and a relatively low synthase activity in the in vitro enzyme assay, which is thought to be the reason for the higher molecular weights of PHA obtained in this study.

Thauera aminoaromatica MZ1T Identified as a Polyhydroxyalkanoate-Producing Bacterium within a Mixed Microbial Consortium.

Polyhydroxyalkanoates (PHAs) form a highly promising class of bioplastics for the transition from fossil fuel-based plastics to bio-renewable and biodegradable plastics. Mixed microbial consortia (MMC) are known to be able to produce PHAs from organic waste streams. Knowledge of key-microbes and their characteristics in PHA-producing consortia is necessary for further process optimization and direction towards synthesis of specific types of PHAs. In this study, a PHA-producing mixed microbial consortium (MMC) from an industrial pilot plant was characterized and further enriched on acetate in a laboratory-scale selector with a working volume of 5 L. 16S-rDNA microbiological population analysis of both the industrial pilot plant and the 5 L selector revealed that the most dominant species within the population is Thauera aminoaromatica MZ1T, a Gram-negative beta-proteobacterium belonging to the order of the Rhodocyclales. The relative abundance of this Thauera species increased from 24 to 40% after two months of enrichment in the selector-system, indicating a competitive advantage, possibly due to the storage of a reserve material such as PHA. First experiments with T. aminoaromatica MZ1T showed multiple intracellular granules when grown in pure culture on a growth medium with a C:N ratio of 10:1 and acetate as a carbon source. Nuclear magnetic resonance (NMR) analyses upon extraction of PHA from the pure culture confirmed polyhydroxybutyrate production by T. aminoaromatica MZ1T.