Polyhydroxybutyrate Research Articles

Surface modification of polyester films with polyfunctional amines: Effect on bacterial biofilm formation

The development of materials with antifouling properties is crucial in many areas, including medicine and food packaging. In this study, 2D-matrices made of polylactic acid (PLA), polyhydroxybutyrate (PHB), or polyhydroxybutyrate-co-valerate (PHB-HV) were surface functionalized through aminolysis with three polyfunctional amines, 1,6-hexamethylenediamine (HDA), tetraethylenepentamine (TEPA), and polyallylamine hydrochloride (PAH). The aminolysis procedure was thoroughly studied to ensure a high amount of amine groups while preserving the structural properties of the films. Interestingly, PHB and PHB-HV were found to be more sensitive to aminolysis than PLA, and the highest amino group density was achieved in surfaces etched with PAH. A decrease in the contact angle from ca. 85° to ca. 70° was revealed for polymers functionalized with HDA and TEPA and a drastic reduction in Staphylococcus epidermidis adhesion was observed for PHB-HV functionalized with the polymeric amine PAH. Polymer antimicrobial activity was found to be related to the degree of surface functionalization. The functionalized, cationic polymer surfaces were supposed to act upon contact with bacteria, without releasing any antimicrobial agent. The developed bioactive surfaces may have potential applications as flexible films for food packaging.

Structure and Performance of All-Green Electrospun PHB-Based Membrane Fibrous Biomaterials Modified with Hemin

This work addresses the challenges concerning the development of "all-green" high-performance biodegradable membrane materials based on poly-3-hydroxybutyrate (PHB) and a natural biocompatible functional additive, iron-containing porphyrin, Hemin (Hmi) via modification and surface functionalization. A new facile and versatile approach based on electrospinning (ES) is advanced when modification of the PHB membranes is performed by the addition of low concentrations of Hmi (from 1 to 5 wt.%). Structure and performance of the resultant {HB/Hmi membranes were studied by diverse physicochemical methods, including differential scanning calorimetry, X-ray analysis, scanning electron microscopy, etc. Modification of the PHB fibrous membranes with Hmi allows control over their quality, supramolecular structure, morphology, and surface wettability. As a result of this modification, air and liquid permeability of the modified electrospun materials markedly increases. The proposed approach provides preparation of high-performance all-green membranes with tailored structure and performance for diverse practical applications, including wound healing, comfort textiles, facial protective masks, tissue engineering, water and air purification, etc.

Biological and Oxidative Degradation of Ultrathin-Fibrous Nonwovens Based on Poly(lactic Acid)/Poly(3-Hydroxybutyrate) Blends.

Developing biodegradable materials based on polymer blends with a programmable self-destruction period in the environmental conditions of living systems is a promising direction in polymer chemistry. In this work, novel non-woven fibrous materials obtained by electrospinning based on the blends of poly(lactic acid) (PLA) and poly(3-hydroxybutyrate) (PHB) were developed. The kinetics of biodegradation was studied in the aquatic environment of the inoculum of soil microorganisms. Oxidative degradation was studied under the ozone gaseous medium. The changes in chemical composition and structure of the materials were studied by optical microscopy, DSC, TGA, and FTIR-spectroscopy. The disappearance of the structural bands of PHB in the IR-spectra of the blends and a significant decrease in the enthalpy of melting after 90 days of exposure in the inoculum indicated the biodegradation of PHB while PLA remained stable. It was shown that the rate of ozonation was higher for PLA and the blends with a high content of PLA. The lower density of the amorphous regions of the blends determined an increased rate of their oxidation by ozone compared to homopolymers. The optimal composition in terms of degradation kinetics is a fibrous material based on the blend of 30PLA/70PHB that can be used as an effective ecosorbent, for biopackaging, and as a highly porous covering material for agricultural purposes.

Non-sterilized conversion of whole lignocellulosic components into polyhydroxybutyrate by Halomonas sp. Y3 with a dual anti-microbial contamination system

Polyhydroxybutyrate (PHB) production from lignocellulosic biomass is challenging due to the need for whole components and energy-effective conversion. Herein, Halomonas sp. Y3, a ligninolytic bacterium with the capacity to produce PHB from lignin and cellulose- and hemicellulose-derived sugars, is employed to explore its feasibility. This strain shows high sugar tolerance up to 200 g/L of glucose and 120 g/L of xylose. A dual anti-microbial contamination system (DACS) containing alkali-halophilic system (AHS) and phosphite-urea system (PUS) is presented, successfully achieving a completely aseptic effect and resulting in a total of 8.2 g of PHB production from 100 g bamboo biomass. We further develop a stage-fed-batch fermentation to promote the complete utilization of xylose. Approximately 69.99 g of dry cell weight (DCW) and 46.45 g of PHB with 66.35 % are obtained from a total of 296.58 g of sugars and 5.70 g of lignin, showing a significant advancement for LCB bioconversion. We then delete the native phosphate transporters, rendering the strain unable to grow on phosphate-loaded media, effectively improving the strain biosafety without compromising its ability to produce PHB. Overall, our findings demonstrate the potential of Y3 as a classic bacterium strain for PHB production with potential uses in industry.

Construction of Stretchable and Large Deformation Green Triboelectric Nanogenerator and Its Application in Technical Action Monitoring of Racket Sports

The rapid development of E-Skin has attracted researchers’ attention to portable wearable devices. However, it is a great challenge to achieve high-output performance and high-environmental protection simultaneously. Polyhydroxybutyrate (PHB) has excellent electron-withdrawing ability and biodegradability, but its stretching ability is poor, which limits its application in portable wearable devices. In this paper, the biocompatible poly(lactide-co-caprolactone) (PLCL) was innovatively selected to modify the properties of the PHB electrospun membranes, and PHB/PLCL composite membranes with different contents of PLCL were constructed. Then, the expanded polytetrafluoroethylene (ePTFE) membranes and PHB/PLCL electrospun membranes were used as the friction layers to construct a flexible triboelectric nanogenerator (TENG). With the increase of PLCL contents, the elastic recovery rate of the composite membranes increased to 68% of tension at 40% strain. The toughness of the composite membrane was increased by 546%, which exhibits ultratoughness and stretchable properties. Based on the ultratoughness and high elasticity electrospun membranes with 50% PLCL, a stretchable and large deformation sensing device was made. The PHB/PLCL-ePTFE TENG could distinguish pingpong and badminton accurately, as well as their various postures such as forehand, backhand, smash poses, etc., which provided a new idea for large deformation monitoring of TENGs.

Assessing the Biodegradability of PHB-Based Materials with Different Surface Areas: A Comparative Study on Soil Exposure of Films and Electrospun Materials.

Due to the current environmental situation, biopolymers are replacing the usual synthetic polymers, and special attention is being paid to poly-3-hydroxybutyrate (PHB), which is a biodegradable polymer of natural origin. In this paper, the rate of biodegradation of films and fibers based on PHB was compared. The influence of exposure to soil on the structure and properties of materials was evaluated using methods of mechanical analysis, the DSC method and FTIR spectroscopy. The results showed rapid decomposition of the fibrous material and also showed how the surface of the material affects the rate of biodegradation and the mechanical properties of the material. It was found that maximum strength decreased by 91% in the fibrous material and by 49% in the film. Additionally, the DSC method showed that the crystallinity of the fiber after exposure to the soil decreased. It was established that the rate of degradation is influenced by different factors, including the surface area of the material and its susceptibility to soil microorganisms. The results obtained are of great importance for planning the structure of features in the manufacture of biopolymer consumer products in areas such as medicine, packaging, filters, protective layers and coatings, etc. Therefore, an understanding of the biodegradation mechanisms of PHB could lead to the development of effective medical devices, packaging materials and different objects with a short working lifespan.

Valorization of Waste from Argan Seeds for Polyhydroxybutyrate Production Using Bacterial Strains Isolated from Argan Soils.

The aim of this work was to study the valorization of argan seed pulp, a waste material obtained from argan oil extraction, for the biosynthesis of polyhydroxybutyrate (PHB). A new species that showed the metabolic capacity for the conversion of argan waste into the bio-based polymer was isolated from an argan crop located in Teroudant, a southwestern region of Morocco, where the arid soil is exploited for goat grazing. The PHB accumulation efficiency of this new species was compared to the previously identified species 1B belonging to the genus Sphingomonas, and results were reported as dry cell weight residual biomass and PHB final yield measured. Temperature, incubation time, pH, NaCl concentration, nitrogen sources, residue concentrations, and culture medium volumes were analyzed with the aim of obtaining a maximum accumulation of PHB. UV-visible spectrophotometry and FTIR analysis confirmed that PHB was present in the material extracted from the bacterial culture. The results of this wide investigation indicated that the new isolated species 2D1 had a higher efficiency in PHB production compared to the previously identified strain 1B, which was isolated from a contaminated argan soil in Teroudant. PHB final yield of the two bacterial species, i.e., the new isolated and 1B, cultivated under optimal culture conditions, in 500 mL MSM enriched with 3% argan waste, were 21.40% (5.91 ± 0.16 g/L) and 8.16% (1.92 ± 0.23 g/L), respectively. For the new isolated strain, the result of the UV-visible spectrum indicates the absorbance at 248 nm, while the FTIR spectrum showed peaks at 1726 cm-1 and 1270 cm-1: these characteristic peaks indicated the presence of PHB in the extract. The data from the species 1B UV-visible and FTIR spectra were previously reported and were used in this study for a correlation analysis. Furthermore, additional peaks, uncharacteristic of standard PHB, suggest the presence of impurities (e.g., cell debris, solvent residues, biomass residues) that persisted after extraction. Therefore, a further enhancement of the sample purification during extraction is recommended for more accuracy in the chemical characterization. If 470,000 tons of argan fruit waste can be produced annually, and 3% of waste is consumed in 500 mL culture by 2D1 to produce 5.91 g/L (21.40%) of the bio-based polymer PHB, it can be estimated that the amount of putative PHB that can be extracted annually from the total argan fruit waste is about 2300 tons.

Biodegradation Studies of Polyhydroxybutyrate and Polyhydroxybutyrate-co-Polyhydroxyvalerate Films in Soil.

Due to increased environmental pressures, significant research has focused on finding suitable biodegradable plastics to replace ubiquitous petrochemical-derived polymers. Polyhydroxyalkanoates (PHAs) are a class of polymers that can be synthesized by microorganisms and are biodegradable, making them suitable candidates. The present study looks at the degradation properties of two PHA polymers: polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-polyhydroxyvalerate (PHBV; 8 wt.% valerate), in two different soil conditions: soil fully saturated with water (100% relative humidity, RH) and soil with 40% RH. The degradation was evaluated by observing the changes in appearance, chemical signatures, mechanical properties, and molecular weight of samples. Both PHB and PHBV were degraded completely after two weeks in 100% RH soil conditions and showed significant reductions in mechanical properties after just three days. The samples in 40% RH soil, however, showed minimal changes in mechanical properties, melting temperatures/crystallinity, and molecular weight over six weeks. By observing the degradation behavior for different soil conditions, these results can pave the way for identifying situations where the current use of plastics can be replaced with biodegradable alternatives.

Taguchi-grey relational analysis in parameter optimisation of green biopolymer composites

ABSTRACT This study investigated the mechanical properties of blends and their nanocomposite blends (BNCs) of polyhydroxybutyrate (PHB) and polylactic acid (PLA) at different loadings (0.5, 1, and 2%) of cellulose nanofibrils (CNF) and nano titanium dioxide (n–TiO2), and parameter optimisation of the BNCs was conducted using a statistical technique such as Taguchi-grey relational and regressional analysis. The results showed that adding PLA to PHB increased the mechanical properties of the blends, and the mechanical properties increased more with the high loading of PLA. The reinforcement with n–TiO2 improved the mechanical properties of the blends; however, the presence of the CNF in the blends generally decreased the mechanical properties. The grey relational analysis showed that the best BNCs were 75% PHB /25% PLA with 0.5% CNF. However, Taguchi analysis found that 75% PHB / 25% PLA with 2% n–TiO2 has the best mechanical properties.

Intracellular carbon storage by microorganisms is an overlooked pathway of biomass growth

The concept of biomass growth is central to microbial carbon (C) cycling and ecosystem nutrient turnover. Microbial biomass is usually assumed to grow by cellular replication, despite microorganisms’ capacity to increase biomass by synthesizing storage compounds. Resource investment in storage allows microbes to decouple their metabolic activity from immediate resource supply, supporting more diverse microbial responses to environmental changes. Here we show that microbial C storage in the form of triacylglycerides (TAGs) and polyhydroxybutyrate (PHB) contributes significantly to the formation of new biomass, i.e. growth, under contrasting conditions of C availability and complementary nutrient supply in soil. Together these compounds can comprise a C pool 0.19 ± 0.03 to 0.46 ± 0.08 times as large as extractable soil microbial biomass and reveal up to 279 ± 72% more biomass growth than observed by a DNA-based method alone. Even under C limitation, storage represented an additional 16–96% incorporation of added C into microbial biomass. These findings encourage greater recognition of storage synthesis as a key pathway of biomass growth and an underlying mechanism for resistance and resilience of microbial communities facing environmental change.

Antibiotic resistance genes proliferation under anaerobic degradation of polylactic acid and polyhydroxy butyrate bioplastics

As the global concern over plastic pollution grows, efforts are underway to find environmentally friendly alternatives to traditional plastics. Bioplastics are being extensively researched and developed as a possible solution. This study compared the impact of two bioplastics, polylactic acid (PLA) and polyhydroxy butyrate (PHB), on the proliferation of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) during anaerobic digestion (AD). Both bioplastics (250–500 particles) could be degraded to a certain extent over 79 days, as indicated by higher methane production than the control without bioplastic particles. The PHB 500 reactor showed the highest methane yield along with the highest biodegradation efficiency (91 %) than other reactors amended with PHB and PLA particles. The highest ARG and MGE abundances were also observed in PLA 500, and the lowest ARG abundance was in PLA 250. Conversely, PHB reactors showed a relatively lower ARG abundance than the control. The correlation analysis suggested that most ARGs were positively correlated with PLA and negatively correlated with PHB (except for tetA, tetB, and tetX). Moreover, a correlation between MGEs and ARGs in PLA and PHB reactors was revealed by correlation analysis. These results show that AD responds differently to the different types/levels of bioplastics, which can ultimately influence the behavior of ARG proliferation. Thus, bioplastics may also pose a potential risk for spreading antibiotic resistance. These findings can be used as a basis for setting environmental standards for bioplastics and creating monitoring and control measures to prevent potential negative impacts on public health.

Isolation and Screening of Polyhydroxybutyrate (PHB) Producing Bacteria from Soils

Isolation and Screening of Polyhydroxybutyrate (PHB) Producing Bacteria from Soils

PHAs production by facultative anaerobic bacteria Bacillus cereus FM5 through submerged and solid-state fermentation under anoxic condition.

PHAs (polyhydroxyalkanoates) are the bio-polyester synthesized by different aerobic and anaerobic bacteria as energy storage granule. However, its synthesis by anaerobes or facultative anaerobes is an imperative part of their physiology via assimilating broad range of substrates than aerobes. Thus, three Gram positive facultative anaerobic PHAs producers viz., Enterococcus sp. FM3, Actinomyces sp. CM4 and Bacillus sp. FM5 were selected. Among them, Bacillus sp. FM5 showed higher cell biomass production in MSM (mineral salt medium) comprised of glucose & peptone as carbon & nitrogen source at pH 9, temperature 37°C, inoculum 10% and incubation period 72h. Under optimized condition, Bacillus sp. FM5 produced 0.89 and 1.5gl-1 of PHAs through submerged and solid-state fermentation in anoxic condition. In-silico analysis confirmed the facultative anaerobic PHAs producing bacteria as Bacillus cereus FM5. IR spectra of PHAs illustrated a strong absorption peak at 1718.50cm-1 representing carbonyl ester (C=O) functional group of PHB (polyhydroxybutyrate), belonging to the family PHAs. It is the first report demonstrating PHAs production by Bacillus cereus FM5 in anoxic condition through different bioprocess technology, which may pave the way in the arena of further biopolymer research.

Life cycle assessment and mechanical properties of nanocomposites based on cellulose nanocrystals

AbstractThe present work focuses on the preparation of cellulose nanocrystals (CNCs) reinforced composites based on polyhydroxybutyrate (PHB) and polypropylene (PP). The effects of the CNCs content as reinforcement and the addition of maleic anhydride (MA) as a coupling agent on the mechanical and morphological properties of the composites were studied. In addition, the biodegradability of the composites was investigated. Samples were prepared using PP and PHB as matrixes and CNCs as reinforcement. Test samples were prepared using the injection molding machine. The studied parameters were microscopic analysis and tensile strength measurement. Tensile strength increased with increasing CNC, but it decreased slightly when 5 wt% of CNC was added. Tensile strength decreased with the addition of MA. Environmental impacts from the production of 100 g of PP and PHB were evaluated using the life cycle assessment (LCA) approach. The results showed that the environmental impacts of composites containing PHB biopolymer are higher than PP ones.

Paraburkholderia tagetis sp. nov., a novel species isolated from roots of Tagetes patula enhances the growth and yield of Solanum lycopersicum L. (tomato).

A multifunctional, Gram-stain-negative, aerobic, motile by flagella, short-rod shaped bacteria, designated strain RG36T was isolated from roots of marigold plant (Tagetes patula) sampled at Dongguk University, Republic of Korea. A 16S rRNA sequences indicated that the closest phylogenetic neighbors were Paraburkholderia acidiphila 7Q-K02T (99.0%) and Paraburkholderia sacchari IPT101T (98.9%) of the family Burkholderiaceae. The draft genome size was 8.52 Mb (63.7% GC). The genome contained 7,381 coding sequences. Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values of strain RG36T with its most closely related species were only 83.1-88.7 and 27.6-36.7%, respectively. Strain RG36T contained Q-8 as the major respiratory quinone and its main fatty acids (>10%) were C16:0, C17:0 cyclo, C19:0 cyclo ω8c, and summed feature 8 (comprising C18:1 ω7c and/or C18:1 ω6c). Strain RG36T accumulates polyhydroxybutyrates (PHB) and exhibits multiple plant growth-promoting properties including production of indole-3-acetic acid (IAA), siderophores, protease, phosphate solubilization, and harboring gene clusters for its multifunctional properties. A pot experiment was conducted to evaluate the effect of PGPR on the growth of Solanum lycopersicum L. (Tomato). Result also confirmed the ability of strain RG36T to promote tomato plant growth, especially it increases the yield of tomatoes. Structural assessment of the bioplastic by Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR), and GC-MS spectroscopy, which confirmed the structure of the polymer as PHB. Our study revealed the potential of strain RG36T to promote the growth of tomato plant and fruit yield by stimulating the various phytohormones, which could be use as bio-fertilizers to reduce the use of chemical fertilizers and promotes sustainable agricultural production. The phenotypic, chemotaxonomic and phylogenetic data, and genome analysis showed that strain RG36T represents a novel species of the genus Paraburkholderia, for which the name Paraburkholderia tagetis sp. nov. is proposed. The type strain is RG36T (=KACC 22685T = TBRC 15696T).

Development of biopolymers from microbes and their environmental applications

Abstract Inventions begin with the invasion of humans and furnish a better livelihood. In some cases, it turns out to be imperative. The environmental issues of using synthetic polymers, including bio-incompatibility, toxicity, high cost, poor hydrophilicity, and pro-inflammatory degradation of byproducts, are increasing the need for and application of eco-friendly, alternative polymeric substances from medicine to biotechnology, which includes the industries of medicine, cosmetics, confectionery, wastewater treatment, etc., as tissue scaffolds, wound dressings, drug packaging material, dermal fillers, moisturising cream, carriers, sun protectants, antiperspirants, and deodorants; gelling agents; stabilisers, emulsifiers, photographic films, etc. Biopolymers are available in different compounds, produced by microbes, plants, and animals, where microbes, for example, Pseudomonas aeruginosa and Kamagataeibacter sucrofermetans, retain these compounds at an exorbitant level, helping them to sustain adverse conditions. Moreover, compared to plant and animal biopolymers, microbial biopolymers are preferred due to their ease of production, design, and processing at an industrial levels. In this regard, polyhydroxyalkanoates (PHA) and poly-3-hydroxybutyrate (PHB) have together attained assiduity for their biodegradable properties and possess similar features as petrochemical-based polymers, commonly synthetic polymers like polyethylene, polypropylene, etc. This attributes to its non-toxic nature, i.e., it behaves eco-friendly by degrading the components through a carbon-neutral energy cycle to carbon dioxide and water, which lessens the dependence on petroleum-based polymers. This chapter contemplates the methods to develop biopolymers from microbes and their environmental applications, focusing on the confiscation of heavy metals, organic dyes or oils, etc.

Increased Biomass and Polyhydroxybutyrate Production by Synechocystis sp. PCC 6803 Overexpressing RuBisCO Genes.

The overexpression of the RuBisCO (rbc) gene has recently become an achievable strategy for increasing cyanobacterial biomass and overcoming the biocompound production restriction. We successfully constructed two rbc-overexpressing Synechocystis sp. PCC 6803 strains (OX), including a strain overexpressing a large subunit of RuBisCO (OXrbcL) and another strain overexpressing all large, chaperone, and small subunits of RuBisCO (OXrbcLXS), resulting in higher and faster growth than wild type under sodium bicarbonate supplementation. This increased biomass of OX strains significantly contributed to the higher polyhydroxybutyrate (PHB) production induced by nutrient-deprived conditions, in particular nitrogen (N) and phosphorus (P). As a result of higher PHB contents in OX strains occurring at days 7 and 9 of nutrient deprivation, this enhancement was apparently made possible by cells preferentially maintaining their internal lipids while accumulating less glycogen. The OXrbcLXS strain, with the highest level of PHB at about 39 %w/dry cell weight (DCW) during 7 days of BG11-NP treatment, contained a lower glycogen level (31.9 %w/DCW) than wild type control (40 %w/DCW). In contrast, the wild type control strain exposed to N- and NP-stresses tended to retain lipid levels and store more glycogen than PHB. In this model, we, for the first time, implemented a RuBisCO-overexpressing cyanobacterial factory for overproducing PHB, destined for biofuel and biomaterial biotechnology.

Structure analysis and thermal stability of PHB recovered from sugar industry waste

ABSTRACT The current article emphasized on cultural conditions for Alcaligenes sp. NCIM 5085 to synthesize Polyhydroxybutyrate (PHB) from sugar industry waste during batch fermentation. Alcaligenes sp. NCIM 5085 was found to grow best in conditions that included 40 g l−1 of cane molasses, 1 g l−1 of ammonium sulphate, 10% inoculum with neutral pH and 48 h of incubation time. Sudan Black B staining was employed to verify the PHB synthesis initially, and further TEM analysis was performed to confirm it. The structural analysis of recovered PHB was carried out by using GC-MS, FTIR, 1H NMR and 13C NMR analysis. The absorption peak at 1724.56 cm−1 revealed the presence of C=O (carbonyl) group by FTIR, which is an indicator of PHB presence. Furthermore, results of NMR and GC-MS analysis confirmed the recovered polymer was PHB. The thermal properties of recovered polymer were analyzed by TGA, DTG and DSC and showed thermal stability of PHB. The observed glass transient temperature (Tg) −2.8°C was within the normal PHB range of Tg. However, melting temperature of recovered PHB was 161.7°C, where the degree of crystallinity was lower than standard PHB that widens the application possibilities.

A solvent-free process to design low density polyhydroxyalkanoate

The polyhydroxyalkanoates (PHAs) are biobased, biocompatible and biodegradable polyesters. To enhance their biodegradability and flexibility, we develop low density PHAs by an emulsion-templated method without organic solvent. Polycaprolactone (PCL) and castor oil are required to improve the flexibility and to generate the porosity, respectively. To overcome the well-known lack of compatibility between PHA and PCL, pluronic is introduced to improve the miscibility of these polyesters The densities of native PHA and PCL are 1.2 g.cm−3 and the densities decreased to 0.49 g.cm−3 in presence of castor oil (40 wt%). Different PHAs were studied: polyhydroxybutyrate (PHB), polyhydroxybutyrate-co-hydroxyvalerate (PHBHV) and polyhydroxybutyrate-co-hydroxyhexanoate (PHBHHx). The porous PHBHHx based material showed the best colonization by Pseudomonas, followed by the PHBHV and PHB. This colonization only occurred in the surface and no bacterial diffusion was observed inside the material. The biodegradability study in presence of lipase showed that after 8 days, the weight losses are, respectively, 20% and 75% when the densities decreased from 0.82 g.cm−3 to 0.49 g.cm−3. These results showed the importance of the porosity on the biodegradation of PHAs.

Acylated plastoquinone is a novel neutral lipid accumulated in cyanobacteria.

Although cyanobacteria do not possess bacterial triacylglycerol (TAG)-synthesizing enzymes, the accumulation of TAGs and/or lipid droplets has been repeatedly reported in a wide range of species. In most cases, the identification of TAG has been based on the detection of the spot showing the mobility similar to the TAG standard in thin-layer chromatography (TLC) of neutral lipids. In this study, we identified monoacyl plastoquinol (acyl PQH) as the predominant molecular species in the TAG-like spot from the unicellular Synechocystis sp. PCC 6803 (S.6803) as well as the filamentous Nostocales sp., Nostoc punctiforme PCC 73102, and Anabaena sp. PCC 7120. In S.6803, the accumulation level of acyl PQH but not TAG was affected by deletion or overexpression of slr2103, indicating that acyl PQH is the physiological product of Slr2103 having homology with the eukaryotic diacylglycerol acyltransferase-2 (DGAT2). Electron microscopy revealed that cyanobacterial strains used in this study do not accumulate lipid droplet structures such as those observed in oleaginous microorganisms. Instead, they accumulate polyhydroxybutyrate (PHB) granules and/or aggregates of alkane, free C16 and C18 saturated fatty acids, and low amounts of TAG in the cytoplasmic area, which can be detected by staining with a fluorescent dye specific to neutral lipids. Unlike these lipophilic materials, acyl PQH is exclusively localized in the membrane fraction. There must be DGAT2-like enzymatic activity esterifying de novo-synthesized C16 and C18 fatty acids to PQH2 in the thylakoid membranes.