Radiation induced graft copolymerization of acrylamide onto poly (3-hydroxybutyrate)

International Nuclear Information System (INIS)

Gonzalez Torres, Maykel; Rapado Paneque, Manuel; Paredes Zaldivar, Mayte; Altanes Valentin, Sonia; Barrera Gonzalez, Gisela

2008-01-01

The graft copolymer poly (3-hydroxybutyrate)-g- polyacrylamide [P (HB-g-AAm)] was synthesized by radiation induced graft copolymerization of acrylamide onto poly (3-hydroxybutyrate). The study was conducted by the simultaneous irradiation method. The structure of [P (HB-g-AAm)] was identified by Fourier Transform Infrared (FTIR) spectroscopy. Thermal behavior of the graft copolymer was also studied by Thermal Gravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC). From the results it was found that FTIR studies showed new signals (stretching -N-H) as strong evidence of grafting. The grafting degree was found to be 10 % and the thermodynamic parameter obtained from the DSC thermogram of plain PHB and the graft copolymer varied showing decrease in the material crystallinity and increase in the glass transition temperature. These results demonstrate that the radiation induced graft copolymerization reaction of acrylamide onto PHB was successively achieved. (Author)

3D FT-IR imaging spectroscopy of phase-separation in a poly(3-hydroxybutyrate)/poly(L-lactic acid) blend

Science.gov (United States)

Miriam Unger; Julia Sedlmair; Heinz W. Siesler; Carol Hirschmugl; Barbara Illman

2014-01-01

In the present study, 3D FT-IR spectroscopic imaging measurements were applied to study the phase separation of a poly(3-hydroxybutyrate) (PHB)/poly(L-lactic acid) (PLA) (50:50 wt.%) polymer blend film. While in 2D projection imaging the z-dependent information is overlapped, thereby complicating the analysis, FT-IR spectro-micro-tomography,...

Synthesis, Characterization and Biocompatibility of Biodegradable Elastomeric Poly(ether-ester urethane)s Based on Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) and Poly(ethylene glycol) via Melting Polymerization

DEFF Research Database (Denmark)

Li, Zibiao; Yang, Xiaodi; Wu, Linping

2009-01-01

Poly(ether-ester urethane)s (PUs) multiblock co-polymers were synthesized from telechelic hydroxylated poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) and poly(ethylene glycol) (PEG) via a melting polymerization (MP) process using 1,6-hexamethylene diisocyanate (HDI) as a non-toxic couplin...

Biodegradable block poly(ester-urethane)s based on poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymers.

Science.gov (United States)

Ou, Wenfeng; Qiu, Handi; Chen, Zhifei; Xu, Kaitian

2011-04-01

A series of block poly(ester-urethane)s (abbreviated as PU3/4HB) based on biodegradable poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3/4HB) segments were synthesized by a facile way of melting polymerization using 1,6-hexamethylene diisocyanate (HDI) as the coupling agent and stannous octanoate (Sn(Oct)(2)) as catalyst, with different 4HB contents and segment lengths. The chemical structure, molecular weight and distribution were systematically characterized by (1)H nuclear magnetic resonance spectrum (NMR), Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC). The thermal property was studied by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). The hydrophilicity was investigated by static contact angle of deionized water and CH(2)I(2). DSC curves revealed that the PU3/4HB polyurethanes have their T(g) from -25.6 °C to -4.3 °C, and crystallinity from 2.5% to 25.3%, being almost amorphous to semi-crystalline. The obtained PU3/4HBs are hydrophobic (water contact angle 77.4°-95.9°), and their surface free energy (SFE) were studied. The morphology of platelets adhered on the polyurethane film observed by scanning electron microscope (SEM) showed that platelets were activated on the PU3/4HB films which would lead to blood coagulation. The lactate dehydrogenase (LDH) assay revealed that the PU3/4HBs displayed higher platelet adhesion property than raw materials and biodegradable polymer polylactic acid (PLA) and would be potential hemostatic materials. Crystallinity degree, hydrophobicity, surface free energy and urethane linkage content play important roles in affecting the LDH activity and hence the platelet adhesion. CCK-8 assay showed that the PU3/4HB is non-toxic and well for cell growth and proliferation of mouse fibroblast L929. It showed that the hydrophobicity is an important factor for cell growth while 3HB content of the PU3/4HB is important for the cell proliferation. Through changing the

Evaluation of 3-hydroxybutyrate as an enzyme-protective agent against heating and oxidative damage and its potential role in stress response of poly(3-hydroxybutyrate) accumulating cells

Czech Academy of Sciences Publication Activity Database

Obruča, S.; Sedláček, P.; Mravec, F.; Samek, Ota; Márová, I.

2016-01-01

Roč. 100, č. 3 (2016), s. 1365-1376 ISSN 0175-7598 R&D Projects: GA MŠk(CZ) LO1212; GA ČR(CZ) GA15-20645S Institutional support: RVO:68081731 Keywords : poly(3-hydroxybutyrate) * 3-Hydroxybutyrate * PHB cycle * chemical chaperone * compatible solutes Subject RIV: BH - Optics, Masers, Lasers Impact factor: 3.420, year: 2016

Magnesium Affects Poly(3-hydroxybutyrate-co-4-hydroxybutyrate Content and Composition by Affecting Glucose Uptake in Delftia acidovorans

Directory of Open Access Journals (Sweden)

Lee, W. H.

2007-01-01

Full Text Available Precise control of polyhydroxyalkanoate (PHA composition is necessary in order to synthesize polymers with specific properties. Among the various types of PHA that have been identified, those that contain 4-hydroxybutyrate (4HB monomers are especially useful in the medical and pharmaceutical fields as absorbable biomaterial. In this study, we have investigated the effect of magnesium concentration on the biosynthesis of poly(3-hydroxybutyrate-co-4-hydroxybutyrate [P(3HB-co-4HB] by Delftia acidovorans DS-17. Our results show that, magnesium affects the copolymer content and composition by affecting glucose uptake from the culture medium. Higher concentrations of magnesium resulted in lower molar fractions of 3HB in the copolymer and reduced uptake of glucose. The results show for the first time that magnesium may be used to achieve fine control of biologically synthesized PHA copolymer composition.

Microbial production and characterization of poly-3-hydroxybutyrate by Neptunomonas antarctica

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Xiao-Jie Liu

2016-08-01

Full Text Available Considering the industrial interest of biodegradable polymer poly-3-hydroxybutyrate (PHB, the marine bacteria Neptunomonas antarctica was studied for its ability to accumulate PHB. The extracted polymer was confirmed to be PHB by nuclear magnetic resonance analysis. In shake flask cultures using natural seawater as medium components, PHB was produced up to 2.12 g/L with a yield of 0.18 g PHB/g fructose. In the presence of artificial seawater, the PHB titer and yield reached 2.13 g/L and 0.13 g PHB/g fructose, respectively. The accumulated polymer gradually decreased when fructose was exhausted, indicating that intracellular PHB was degraded by N. antarctica. The weight-average and number-average molecular weights of PHB produced within natural seawater were 2.4 × 105 g/mol and 1.7 × 105 g/mol, respectively. Our results highlight the potential of N. antarctica for PHB production with seawater as a nutrient source.

Novel extracellular PHB depolymerase from Streptomyces ascomycinicus: PHB copolymers degradation in acidic conditions.

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Javier García-Hidalgo

Full Text Available The ascomycin-producer strain Streptomyces ascomycinicus has been proven to be an extracellular poly(R-3-hydroxybutyrate (PHB degrader. The fkbU gene, encoding a PHB depolymerase (PhaZ Sa , has been cloned in E. coli and Rhodococcus sp. T104 strains for gene expression. Gram-positive host Rhodococcus sp. T104 was able to produce and secrete to the extracellular medium an active protein form. PhaZ Sa was purified by two hydrophobic interaction chromatographic steps, and afterwards was biochemically as well as structurally characterized. The enzyme was found to be a monomer with a molecular mass of 48.4 kDa, and displayed highest activity at 45°C and pH 6, thus being the first PHB depolymerase from a gram-positive bacterium presenting an acidic pH optimum. The PHB depolymerase activity of PhaZ Sa was increased in the presence of divalent cations due to non-essential activation, and also in the presence of methyl-β-cyclodextrin and PEG 3350. Protein structure was analyzed, revealing a globular shape with an alpha-beta hydrolase fold. The amino acids comprising the catalytic triad, Ser(131-Asp(209-His(269, were identified by multiple sequence alignment, chemical modification of amino acids and site-directed mutagenesis. These structural results supported the proposal of a three-dimensional model for this depolymerase. PhaZ Sa was able to degrade PHB, but also demonstrated its ability to degrade films made of PHB, PHBV copolymers and a blend of PHB and starch (7∶3 proportion wt/wt. The features shown by PhaZ Sa make it an interesting candidate for industrial applications involving PHB degradation.

Novel extracellular PHB depolymerase from Streptomyces ascomycinicus: PHB copolymers degradation in acidic conditions.

Science.gov (United States)

García-Hidalgo, Javier; Hormigo, Daniel; Arroyo, Miguel; de la Mata, Isabel

2013-01-01

The ascomycin-producer strain Streptomyces ascomycinicus has been proven to be an extracellular poly(R)-3-hydroxybutyrate (PHB) degrader. The fkbU gene, encoding a PHB depolymerase (PhaZ Sa ), has been cloned in E. coli and Rhodococcus sp. T104 strains for gene expression. Gram-positive host Rhodococcus sp. T104 was able to produce and secrete to the extracellular medium an active protein form. PhaZ Sa was purified by two hydrophobic interaction chromatographic steps, and afterwards was biochemically as well as structurally characterized. The enzyme was found to be a monomer with a molecular mass of 48.4 kDa, and displayed highest activity at 45°C and pH 6, thus being the first PHB depolymerase from a gram-positive bacterium presenting an acidic pH optimum. The PHB depolymerase activity of PhaZ Sa was increased in the presence of divalent cations due to non-essential activation, and also in the presence of methyl-β-cyclodextrin and PEG 3350. Protein structure was analyzed, revealing a globular shape with an alpha-beta hydrolase fold. The amino acids comprising the catalytic triad, Ser(131)-Asp(209)-His(269), were identified by multiple sequence alignment, chemical modification of amino acids and site-directed mutagenesis. These structural results supported the proposal of a three-dimensional model for this depolymerase. PhaZ Sa was able to degrade PHB, but also demonstrated its ability to degrade films made of PHB, PHBV copolymers and a blend of PHB and starch (7∶3 proportion wt/wt). The features shown by PhaZ Sa make it an interesting candidate for industrial applications involving PHB degradation.

Mechanical and Morphological Properties of Poly-3-hydroxybutyrate/Poly(butyleneadipate-co-terephthalate)/Layered Double Hydroxide Nanocomposites

OpenAIRE

Pak, Yen Leng; Bin Ahmad, Mansor; Shameli, Kamyar; Yunus, Wan Md Zin Wan; Ibrahim, Nor Azowa; Zainuddin, Norhazlin

2013-01-01

Nanocomposites of poly-3-hydroxybutyrate/poly(butyleneadipate-co-terephthalate)/layered double hydroxide (PHB/PBAT/LDH) were prepared from a binary blend of PHB/PBAT and stearate-Zn3Al LDH via a solution casting method using chloroform as solvent in this study. The pristine Zn3Al LDH was synthesized from nitrate salts solution at pH 7 by using coprecipitation technique and then was modified by stearate anions surfactant via ion exchange reaction. As a result, the basal spacing of the LDH was ...

[A comparative study of biodegradation kinetics of biopolymer systems based on poly(3-hydroxybutyrate)].

Science.gov (United States)

Boskhomdzhiev, A P; Banartsev, A P; Makhina, T K; Myshkina, V L; Ivanov, E A; Bagrov, D V; Filatova, E V; Iordanskiĭ, A L; Bonartseva, G A

2009-01-01

The aim of this study was to evaluate and to compare of long-term kinetics curves of biodegradation of poly(3-hydroxybutyrate) (PHB), its copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate), and PHB/polylactic acid blend. The total weight loss and the change of average viscosity molecular weight were used as an index of biodegradation degree. The rate of biodegradation was analyzed in vitro in presence oflipase and in vivo when the films were implanted in animal tissues. The morphology of PHB films surface was studied by atomic force microscopy technique. It was shown that biodegradation of PHB is occurred by means of as polymer hydrolysis, and as its enzymatic biodegradation. The obtained data can be used for development of medical devices on the base of PHB.

Poly(3-hydroxybutyrate-co-R-3-hydroxyhexanoate) nanoparticles with polyethylenimine coat as simple, safe, and versatile vehicles for cell targeting

DEFF Research Database (Denmark)

Wu, Linping; Wang, Danyang; Parhamifar, Ladan

2014-01-01

A simple and highly safe poly(3-hydroxybutyrate-co-R-3-hydroxyhexanoate) nanoparticulate delivery system that targets different cell types is developed. A sub-cytotoxic level of polyethylenimine coat mediates universal cell targeting. Internalized nanoparticles traffic along endolysosomal compart...... compartments, endoplasmic reticulum and the Golgi complex. Nanoparticles have no detrimental effects on cell morphology and respiration.......A simple and highly safe poly(3-hydroxybutyrate-co-R-3-hydroxyhexanoate) nanoparticulate delivery system that targets different cell types is developed. A sub-cytotoxic level of polyethylenimine coat mediates universal cell targeting. Internalized nanoparticles traffic along endolysosomal...

Characterization of a Novel Subgroup of Extracellular Medium-Chain-Length Polyhydroxyalkanoate Depolymerases from Actinobacteria

Science.gov (United States)

Gangoiti, Joana; Santos, Marta; Prieto, María Auxiliadora; de la Mata, Isabel; Llama, María J.

2012-01-01

Nineteen medium-chain-length (mcl) poly(3-hydroxyalkanoate) (PHA)-degrading microorganisms were isolated from natural sources. From them, seven Gram-positive and three Gram-negative bacteria were identified. The ability of these microorganisms to hydrolyze other biodegradable plastics, such as short-chain-length (scl) PHA, poly(ε-caprolactone) (PCL), poly(ethylene succinate) (PES), and poly(l-lactide) (PLA), has been studied. On the basis of the great ability to degrade different polyesters, Streptomyces roseolus SL3 was selected, and its extracellular depolymerase was biochemically characterized. The enzyme consisted of one polypeptide chain of 28 kDa with a pI value of 5.2. Its maximum activity was observed at pH 9.5 with chromogenic substrates. The purified enzyme hydrolyzed mcl PHA and PCL but not scl PHA, PES, and PLA. Moreover, the mcl PHA depolymerase can hydrolyze various substrates for esterases, such as tributyrin and p-nitrophenyl (pNP)-alkanoates, with its maximum activity being measured with pNP-octanoate. Interestingly, when poly(3-hydroxyoctanoate-co-3-hydroxyhexanoate [11%]) was used as the substrate, the main hydrolysis product was the monomer (R)-3-hydroxyoctanoate. In addition, the genes of several Actinobacteria strains, including S. roseolus SL3, were identified on the basis of the peptide de novo sequencing of the Streptomyces venezuelae SO1 mcl PHA depolymerase by tandem mass spectrometry. These enzymes did not show significant similarity to mcl PHA depolymerases characterized previously. Our results suggest that these distinct enzymes might represent a new subgroup of mcl PHA depolymerases. PMID:22865072

Biodegradation evaluation of bacterial cellulose, vegetable cellulose and poly (3-hydroxybutyrate in soil

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Suellen Brasil Schröpfer

2015-04-01

Full Text Available In recent years, the inappropriate disposal of polymeric materials has increased due to industrial development and increase of population consumption. This problem may be minimized by using biodegradable polymers, such as bacterial cellulose and poly(hydroxybutyrate, from renewable resources. This work was aimed at monitoring and evaluating degradation of bacterial cellulose, vegetable cellulose and poly(3-hydroxybutyrate using Thermogravimetric Analysis and Scanning Electron Microscopy. Controlled mass polymer samples were buried in pots containing soil. Samples were removed in 30 day intervals up to 180 days. The results show that the mass of the polymer increased in the first month when in contact with the soil but then it was degraded as evidenced by mass loss and changes on the sample surface.

Study on the Effect of Levulinic Acid on Whey-Based Biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate by Hydrogenophaga pseudoflava

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Martin Koller

2017-04-01

Full Text Available Background and Objective: Production of polyhydroxyalkanoate copolyesters consisting of 3-hydroxybutyrate and 3-hydroxyvalerate units was for the first time studied using the production strain Hydrogenophaga pseudoflava based on sustainable raw materials. This strategy provides for increased cost efficiency in PHA production and in enhanced material quality.Material and Methods: As a particularity, production of these poly(3-hydroxybutyrate-co-3- hydroxyvalerate copolyesters was based on a novel substrate/co-substrate combination: whey permeate from dairy industry, on the one hand, acted as substrate for biomass and 3HB biosynthesis; on the other hand, levulinic acid, accessible from various renewable resources, was used as 3HV-related precursor compound. The experiments were carried out on shaking flask scale using defined nutrient media.Results and Conclusion: Applied during nutritionally balanced growth of H. pseudoflava, levulinicacid displays drastic growth inhibition at rather low concentrations of 0.2 g l-1 (growth inhibition constant Ki = 0.032, which suggests the careful supply of this compound in the first phase of cultivation. Under nitrogen-free cultivation conditions, inhibition of the strain´s metabolism by levulinic acid was less pronounced. Here, poly(3-hydroxybutyrate-co- 3-hydroxyvalerate concentrations up to 4.2 g l-1 and volumetric poly(3-hydroxybutyrate-co-3- hydroxyvalerate productivities up to 0.06 g l-1 h -1 were achieved in dependence on the precursor supply. Investigating poly(3-hydroxybutyrate-co-3-hydroxyvalerate composition in setups supplied with differently composed whey/levulinic acid mixtures revealed 3- hydroxyvalerate fractions in the polymer between 0 and 0.6 mol mol-1 . This study successfully demonstrates the feasibility of combined utilization of different waste- and by-products from food industry and agriculture for generation of value-added 2nd generation biopolymers. Conflict of interest: The authors

Methanol-induced chain termination in poly(3-hydroxybutyrate) biopolymers: molecular weight control

Science.gov (United States)

A systematic study was performed to demonstrate the impact of methanol (MeOH) on poly(3-hydroxybutyrate) (PHB) synthesis and molecular weight (MW) control. Glycerine (init. conc. = 1.0%; w/v), was used as the primary carbon source in batch-culture fermentations with varying concentrations (0 to 0.85...

In vitro three-dimensional coculturing poly3-hydroxybutyrate-co-3-hydroxyhexanoate with mouse-induced pluripotent stem cells for myocardial patch application.

Science.gov (United States)

Shijun, Xu; Junsheng, Mu; Jianqun, Zhang; Ping, Bo

2016-03-01

Identifying a suitable polymeric biomaterial for myocardial patch repair following myocardial infarction, cerebral infarction, and cartilage injury is essential. This study aimed to investigate the effect of the novel polymer material, poly3-hydroxybutyrate-co-3-hydroxyhexanoate, on the adhesion, proliferation, and differentiation of mouse-induced pluripotent stem cells in vitro. Mouse-induced pluripotent stem cells were isolated, expanded, and cultured on either two-dimensional or three-dimensional poly3-hydroxybutyrate-co-3-hydroxyhexanoate films (membranes were perforated to imitate three-dimensional space). Following attachment onto the films, mouse-induced pluripotent stem cell morphology was visualized using scanning electron microscopy. Cell vitality was detected using the Cell Counting Kit-8 assay and cell proliferation was observed using fluorescent 4',6-diamidino-2-phenylindole (DAPI) staining. Mouse-induced pluripotent stem cells were induced into cardiomyocytes by differentiation medium containing vitamin C. A control group in the absence of an inducer was included. Mouse-induced pluripotent stem cell survival and differentiation were observed using immunofluorescence and flow cytometry, respectively. Mouse-induced pluripotent stem cells growth, proliferation, and differentiation were observed on both two-dimensional and three-dimensional poly3-hydroxybutyrate-co-3-hydroxyhexanoate films. Vitamin C markedly improved the efficiency of mouse-induced pluripotent stem cells differentiation into cardiomyocytes on poly3-hydroxybutyrate-co-3-hydroxyhexanoate films. Three-dimensional culture was better at promoting mouse-induced pluripotent stem cell proliferation and differentiation compared with two-dimensional culture. © The Author(s) 2016.

Effects of the addiction of nucleating agents on crystallization of poly(3-hydroxybutyrate), PHB;Efeito da adicao de agentes de nucleacao na cristalizacao do poli[(R)-3-hidroxibutirato], PHB

Energy Technology Data Exchange (ETDEWEB)

Sarzi, Ariane C.; Felisberti, Maria I., E-mail: ari_sarzi@hotmail.co [Universidade Estadual de Campinas (UNICAMP), SP (Brazil)

2009-07-01

Poly(3-hydroxybutyrate) is a naturally occurring polymer synthesized by bacteria. Because of its biodegradability, it has attracted much attention from medical researchers. However, the inherent brittleness impedes use of bacterial PHB as an engineering plastic. By using a nucleating agent, it is expected that the nucleation density will increase and the size of spherulites will be reduced, resulting in higher crystallization temperatures if compared with pure PHB. The crystallization behavior of poly(3-hydroxybutyrate) (PHB) induced by poly(ethylene terephthalate) (PET ) and other commercials additives as nucleating agents was investigated by differential scanning calorimetry, polarized optical microscopy and X-ray diffraction. (author)

Poly(3-hydroxybutyrate-co-R-3-hydroxyhexanoate) nanoparticles with polyethylenimine coat as simple, safe, and versatile vehicles for cell targeting: population characteristics, cell uptake, and intracellular trafficking.

Science.gov (United States)

Wu, Lin-Ping; Wang, Danyang; Parhamifar, Ladan; Hall, Arnaldur; Chen, Guo-Qiang; Moghimi, Seyed M

2014-06-01

A simple and highly safe poly(3-hydroxybutyrate-co-R-3-hydroxyhexanoate) nanoparticulate delivery system that targets different cell types is developed. A sub-cytotoxic level of polyethylenimine coat mediates universal cell targeting. Internalized nanoparticles traffic along endolysosomal compartments, endoplasmic reticulum and the Golgi complex. Nanoparticles have no detrimental effects on cell morphology and respiration. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Microbial Degradation Behavior in Seawater of Polyester Blends Containing Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate (PHBHHx

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Hitoshi Sashiwa

2018-01-01

Full Text Available The microbial degradation behavior of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate (PHBHHx and its compound with several polyesters such as poly(butylene adipate-co-telephtharate (PBAT, poly(butylene succinate (PBS, and polylactic acid (PLA in seawater was tested by a biological oxygen demand (BOD method. PHBHHx showed excellent biodegradation in seawater in this study. In addition, the biodegradation rate of several blends was much influenced by the weight ratio of PHBHHx in their blends and decreased in accordance with the decrement of PHBHHX ratio. The surface morphology of the sheet was important factor for controlling the biodegradation rate of PHBHHx-containing blends in seawater.

STUDY ON BIODEGRADABILITY OF POLY (3-HYDROXYBUTYRATE-co-3-HYDROXYVALERATE)/ORGANOPHILIC MONTMORILLONITE NANOCOMPOSITES

Institute of Scientific and Technical Information of China (English)

WANG Shufang; SONG Cunjiang; CHEN Guangxin; LIU Jing; YANG Chao; ZHANG Xihui; GUO Tianying; ZHANG Banghua

2004-01-01

Poly (3-hydroxybutyrate-co-3-hydroxyvalerate)/Organophilic montmorillonite (PHBV/OMMT) nanocomposites were prepared and the biodegradability of the PHBV/OMMT nanocomposites was studied by a cultivation degrading method in soil suspension. The relationship between structure and biodegradability of PHBV/OMMT nanocomposites was investigated. The results showed that the biodegradability of PHBV/OMMT nanocomposites decreased with increasing amount of OMMT and it was related to the number of PHBV degrading microorganisms in degradation environment, the anti-microbial property of OMMT and the degree of crystallinity of the nanocomposites.

PhaP phasins play a principal role in poly-β-hydroxybutyrate accumulation in free-living Bradyrhizobium japonicum

OpenAIRE

Yoshida, Ken-ichi; Takemoto, Yuki; Sotsuka, Takayuki; Tanaka, Kosei; Takenaka, Shinji

2013-01-01

Background Bradyrhizobium japonicum USDA110, a soybean symbiont, is capable of accumulating a large amount of poly-β-hydroxybutyrate (PHB) as an intracellular carbon storage polymer during free-living growth. Within the genome of USDA110, there are a number of genes annotated as paralogs of proteins involved in PHB metabolism, including its biosynthesis, degradation, and stabilization of its granules. They include two phbA paralogs encoding 3-ketoacyl-CoA thiolase, two phbB paralogs encoding ...

POLY(3-HYDROXYBUTYRATE) PRODUCTION BY Cupriavidus necator SUPPLEMENTED WITH MINIEMULSIFIED SOYBEAN OIL

OpenAIRE

Schmidt, M.; Ienczak, J. L.; Quines, L. K.; Zanfonato, K.; Schmidell, W.; Aragão, G. M. F.

2016-01-01

Abstract Studies have shown that the supplementation of vegetable oils, in poly(3-hydroxybutyrate) production, provides an increase in the process productivity, besides inducing lipase activity in the medium. The supplementation with miniemulsified oils could potentialize these results. In this work, the influence of supplementation of the medium with soybean oil, without treatment and miniemulsified, on polymerr production was evaluated. The best moment to supplement the medium and its influ...

Co-Expression of ORFCma with PHB Depolymerase (PhaZCma ) in Escherichia coli Induces Efficient Whole-Cell Biodegradation of Polyesters.

Science.gov (United States)

Lee, Ming-Chieh; Liu, En-Jung; Yang, Cheng-Han; Hsiao, Li-Jung; Wu, Tzong-Ming; Li, Si-Yu

2018-04-01

Whole-cell degradation of polyesters not only avoids the tedious process of enzyme separation, but also allows the degraded product to be reused as a carbon source. In this study, Escherichia coli BL21(DE3) harboring phaZ Cma , a gene encoding poly(3-hydroxybutyrate) (PHB) depolymerase from Caldimonas manganoxidans, is constructed. The extra-cellular fraction of E. coli/pPHAZ exhibits a fast PHB degradation rate where it only took 35 h to completely degrade PHB films, while C. manganoxidans takes 81 h to do the same. The co-expression of ORF Cma (a putative periplasmic substrate binding protein that is within the same operon of phaZ Cma ) further improves the PHB degradation. While 28 h is needed for E. coli/pPHAZ to cause an 80% weight loss in PHB films, E. coli/pORFPHAZ needs only 21 h. Furthermore, it is able to degrade at-least four different polyesters, PHB, poly(lactic acid) (PLA), polycaprolactone (PCL), and poly(butylene succinate-co-adipate) (PBSA). Testing of the time course of 3-hydroxybutyrate concentration and the turbidity of the degradation solutions over time shows that PhaZ Cma has both exo- and endo-enzymatic activity. The whole-cell E. coli/pORFPHAZ can be used for recycling various polyesters while ORF Cma can potentially be a universal element for enhancing the secretion of recombinant protein. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Poly(3-Hydroxybutyrate) Synthesis Genes in Azotobacter sp. Strain FA8

OpenAIRE

Pettinari, M. Julia; Vázquez, Gustavo J.; Silberschmidt, Daniel; Rehm, Bernd; Steinbüchel, Alexander; Méndez, Beatriz S.

2001-01-01

Genes responsible for the synthesis of poly(3-hydroxybutyrate) (PHB) in Azotobacter sp. FA8 were cloned and analyzed. A PHB polymerase gene (phbC) was found downstream from genes coding for β-ketothiolase (phbA) and acetoacetyl-coenzyme A reductase (phbB). A PHB synthase mutant was obtained by gene inactivation and used for genetic studies. The phbC gene from this strain was introduced into Ralstonia eutropha PHB-4 (phbC-negative mutant), and the recombinant accumulated PHB when either glucos...

Production and characterization of bio plastics from potato starch, poly-hydroxybutyrate and poly-hydroxybutyrate-co-valerate

International Nuclear Information System (INIS)

Mendes, Fernanda M.; Curvelo, Antonio A.S.

2009-01-01

This work describes the study of thermoplastic starch (TPS) blends obtained from potato starch (plasticised with glycerol) with biodegradable polymers poly-hydroxybutyrate (PHB) and poly-hydroxybutyrate-co-valerate (PHB V). For this purpose it were developed several formulations with TPS/PHB, TPS/PHB V and TPS/PHB/PHB V prepared by physical mixing and water and glycerol as plasticizers. The amount of glycerol was 30% based on starch (dry basis). The starting materials (starch, PHB and PHB V) were characterized by scanning electron microscopy and X-ray diffraction. Then, it was determined the optimal processing conditions for the samples, performed by using an intensive mixer. The materials were hot pressed to produce the standardized samples employed in the characterizations: mechanical testing (tensile strength), dynamic-mechanical thermal analysis (DMTA), scanning electron microscopy and X-ray diffractometry. (author)

Crystal structures, thermal behaviors, and C-H...O=C hydrogen bondings of poly(3-hydroxyvalerate) and poly(3-hydroxybutyrate) studied by infrared spectroscopy and X-ray diffraction

Czech Academy of Sciences Publication Activity Database

Sato, H.; Ando, Y.; Dybal, Jiří; Iwata, T.; Noda, I.; Ozaki, Y.

2008-01-01

Roč. 41, č. 12 (2008), s. 4305-4312 ISSN 0024-9297 R&D Projects: GA ČR GA203/05/0425 Grant - others:Ministry of Education, Culture, Sports, Science and Technology(JP) MEXT Institutional research plan: CEZ:AV0Z40500505 Keywords : poly(3-hydroxyvalerate) * poly(3-hydroxybutyrate) * infrared spectroscopy Subject RIV: CD - Macromolecular Chemistry Impact factor: 4.407, year: 2008

Thermal depolymerization mechanisms of poly (3-hydroxybutyrate-co-3-hydroxyvalerate)

Institute of Scientific and Technical Information of China (English)

Hengxue Xiang; Xiaoshuang Wen; Xiaohui Miu; Yan Li; Zhe Zhou; Meifang Zhu

2016-01-01

Thermal degradation processes and decomposition mechanisms of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) were investigated by using thermal gravity analysis(TGA), Gel permeation chromatography (GPC), elemental analyzer, pyrolysis-gas chromatography-mass spectrometry (PyGC-MS) and 1H nuclear magnetic resonance (1H NMR). The degradation activation energy was calculated via the dependence of residual mass on isothermal temperature. 1H NMR and PyGC-MS were used to investigate the chemical structure and component proportion of volatile gases and degradation residues which were produced by thermal decomposition, and to infer the process of macromolecular chain scission. Besides, the influence of the factors, such as outfield atmosphere, residual metal ions, on the degradation behaviors of PHBV was also studied. Finally, the PHBV thermal decomposition mechanisms were speculated on the basis of the degradation behaviors of molecular and chemical structure.

Metabolism of poly-β-hydroxybutyric acid in bacteroids of Rhizobium lupini in connection with nitrogen fixation and photosynthesis

International Nuclear Information System (INIS)

Romanov, V.I.; Fedulova, N.G.; Tchermenskaya, I.E.; Shramko, V.I.; Molchanov, M.I.; Kretovich, W.L.

1980-01-01

The darkening of lupin plants grown in a sand culture on a nitrogen-free medium at a stage of initial flowering led to a sharply decreased nitrogen fixation intensity which eventually ceased. Decreased intensity of nitrogen fixation in bacteroids was accompanied by an accumulation of poly-β-hydroxybutyric acid (PHB): in the course of 10-20 h (depending upon temperature) its content increased by 2.5-3.0 times. If, following darkening, the plants were once again exposed to light, an abrupt increase of nitrogen fixation intensity was observed and a simultaneous decrease of PHB content. It has been shown that lupin's exposure to light in 14 CO 2 atmosphere lasting 19 h resulted in the latter's incorporation into PHB, bacteroids and into the entire nodule; these processes developed almost in parallel. During the early period of vegetation growth prior to flowering, the PHB content of bacteroids decreased from 13 14 to 3.4% of dry weight, whereas the intensity of nitrogen fixation was raised. Concurrently increase of the activity of some enzymes connected with the PHB metabolism (aceto-acetyl-CoA-reductase, acetyl-CoA acetyl transferase PHB-depolymerase, (CoA-transferase, of 3-ketoacids) occured. The plants' subsequent ageing and reduction of nitrogen fixation intensity led to a noticeable increase of PHB content and a decrease of the above mentioned enzymes' activity. The specific activity of β-hydroxybutyric dehydrogenase involved with PHB catabolism was high and was maintained at a constant level throughout the entire vegetative period. (orig.)

Biodegradable conductive composites of poly(3-hydroxybutyrate and polyaniline nanofibers: Preparation, characterization and radiolytic effects

Directory of Open Access Journals (Sweden)

2011-01-01

Full Text Available Poly(3-hydroxybutyrate is a biodegradable polyester produced by microorganisms under nutrient limitation conditions. We obtained a biodegradable poly(3-hydroxybutyrate composite having 8 to 55% of chemically in situ polymerized hydrochloric acid-doped polyaniline nanofibers (70-100 nm in diameter. Fourier transform infrared spectroscopy and X-rays diffractometry data did not show evidence of significant interaction between the two components of the nanocomposite, and polyaniline semiconductivity was preserved in all studied compositions. Gamma-irradiation at 25 kGy absorbed dose on the semiconductive composite presenting 28% of doped polyaniline increased its conductivity from 4.6*10-2 to 1.1 S/m, while slightly decreasing its biodegradability. PANI-HCl biodegradation is negligible when compared to PHB biodegradability in an 80 day timeframe. Thus, this unprecedented all-polymer nanocomposite presents, at the same time, semiconductivity and biodegradability and was proven to maintain these properties after gamma irradiation. This new material has many potential applications in biological science, engineering, and medicine.

Raman spectroscopy of poly (3-hydroxybutyrate) modified with poly (vinyl acetate) by radiation- induced copolymerization

International Nuclear Information System (INIS)

Gonzalez, Maykel; Galego Fernandez, Norma; Ortiz del Toro, Pedro; Rapado, Manuel; Paredes

2007-01-01

Poly (3-hydroxybutyrate) (PHB) is an important material used in the field of medicine. However in common conditions, PHB has some deficiencies. It is very brittle and slightly hydrophobic polymer. This somewhat limit its applications. Radiation chemistry can be used to improve its chemical properties. In the present study, the substrate, modified by radiation-induced graft copolymerization with vinyl acetate (VAc), was characterized using FTIR and Raman spectroscopy. FTIR spectroscopy did not reveal any significant bands but Raman spectroscopy revealed the formation of a new band that characterize the material

Efficient utilization of crude glycerol as fermentation substrate in the synthesis of poly(3-hydroxybutyrate) biopolymers

Science.gov (United States)

One refined and 2 crude glycerol samples were utilized to produce poly(3-hydroxybutyrate) (PHB) by Pseudomonas oleovorans NRRL B-14682. Fermentation conditions were determined to efficiently utilize glycerol while maintaining PHB yields. A batch culture protocol including 1% glycerol and an aerati...

Mechanical and Morphological Properties of Poly-3-hydroxybutyrate/Poly(butyleneadipate-co-terephthalate/Layered Double Hydroxide Nanocomposites

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Yen Leng Pak

2013-01-01

Full Text Available Nanocomposites of poly-3-hydroxybutyrate/poly(butyleneadipate-co-terephthalate/layered double hydroxide (PHB/PBAT/LDH were prepared from a binary blend of PHB/PBAT and stearate-Zn3Al LDH via a solution casting method using chloroform as solvent in this study. The pristine Zn3Al LDH was synthesized from nitrate salts solution at pH 7 by using coprecipitation technique and then was modified by stearate anions surfactant via ion exchange reaction. As a result, the basal spacing of the LDH was increased from 8.77 to 24.94 Å after the modification. Intercalated nanocomposites were formed due to the presence of diffraction peak in XRD diffractograms. The infrared spectrum of stearate-Zn3Al LDH exhibited the existence of stearate anions in the synthesized Zn3Al LDH. Mechanical properties with 2 wt% stearate-Zn3Al LDH loading nanocomposites showed 56 wt% improvements in elongation at break compared to those of the blend.

Synthesis of Diblock copolymer poly-3-hydroxybutyrate -block-poly-3-hydroxyhexanoate [PHB-b-PHHx] by a β-oxidation weakened Pseudomonas putida KT2442.

Science.gov (United States)

Tripathi, Lakshmi; Wu, Lin-Ping; Chen, Jinchun; Chen, Guo-Qiang

2012-04-05

Block polyhydroxyalkanoates (PHA) were reported to be resistant against polymer aging that negatively affects polymer properties. Recently, more and more attempts have been directed to make PHA block copolymers. Diblock copolymers PHB-b-PHHx consisting of poly-3-hydroxybutyrate (PHB) block covalently bonded with poly-3-hydroxyhexanoate (PHHx) block were for the first time produced successfully by a recombinant Pseudomonas putida KT2442 with its β-oxidation cycle deleted to its maximum. The chloroform extracted polymers were characterized by nuclear magnetic resonance (NMR), thermo- and mechanical analysis. NMR confirmed the existence of diblock copolymers consisting of 58 mol% PHB as the short chain length block with 42 mol% PHHx as the medium chain length block. The block copolymers had two glass transition temperatures (Tg) at 2.7°C and -16.4°C, one melting temperature (Tm) at 172.1°C and one cool crystallization temperature (Tc) at 69.1°C as revealed by differential scanning calorimetry (DSC), respectively. This is the first microbial short-chain-length (scl) and medium-chain-length (mcl) PHA block copolymer reported. It is possible to produce PHA block copolymers of various kinds using the recombinant Pseudomonas putida KT2442 with its β-oxidation cycle deleted to its maximum. In comparison to a random copolymer poly-3-hydroxybutyrate-co-3-hydroxyhexanoate (P(HB-co-HHx)) and a blend sample of PHB and PHHx, the PHB-b-PHHx showed improved structural related mechanical properties.

Comparative Life Cycle Studies on Poly(3-hydroxybutyrate)-Based Composites as Potential Replacement for Conventional Petrochemical Plastics

NARCIS (Netherlands)

Pietrini, M.; Roes, A.L.; Patel, M.K.; Chiellini, E.

2007-01-01

A cradle-to-grave environmental life cycle assessment (LCA) of a few poly(3-hydroxybutyrate) (PHB) based composites has been performed and was compared to commodity petrochemical polymers. The end products studied are a cathode ray tube (CRT) monitor housing (conventionally produced from high-impact

Photoacoustic Monitoring of Internal Plastification in Poly(3-hydroxybutyrate-co-3-hydroxyvalerate Copolymers: Measurements of Thermal Parameters

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Sanchez Ruben R.

1999-01-01

Full Text Available Basic data on thermophysical properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate copolymers poly(3HB-co-3HV were investigated with the aim of understanding the role of 3-hydroxyvalerate monomeric units (3HV incorporated during random copolymerization. The results show strong evidence that internal plastification is produced by the introduction of 3HV units in the copolymer. It was observed that copolymer thermal conductivity increased approximately linearly with the 3HV content. On the other hand, thermal diffusivity was very sensitive to the change in the copolymer composition showing a sudden rise that attained a saturation plateau. Amplitude-frequency plots indicate that a thermoelastic bending mechanism is operating. In this paper a new photoacoustic arrangement for the measurement of thermal effusivity is presented.

High cell density strategy for poly(3-hydroxybutyrate production by Cupriavidus necator

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J. L. Ienczak

2011-12-01

Full Text Available Poly(3-hydroxybutyrate (P(3HB is a carbon and intracellular storage source for different microorganisms and its production can achieve high productivities by means of high cell density cultures. The aim of this study was to propose a high cell density strategy for P(3HB production by Cupriavidus necator. The exponential growth phase demands an accurate control of the oxygen transfer system in the bioreactor, due to maximum specific growth rate (µXr, and, consequently, a maximum specific oxygen uptake rate (QO2, in addition to significant residual biomass (Xr growth in high cell density cultures. In this context, this work investigated the strategy for obtaining high cell density, with the inclusion of a linear growth phase for P(3HB production by C. necator in a fed-batch culture. The linear growth phase was included between the exponential growth phase and the P(3HB production phase as a strategy to reduce the specific growth rate (µXr and specific oxygen uptake rate (QO2, with constant residual biomass growth rate (d(V.Xr/dt = k = constant and linear increase of biomass. Three strategies of culture were performed. The results showed that a high residual biomass concentration (30 gXr.L-1 can be reached by the inclusion of the linear growth strategy and specific growth rates (µXr between 0.08 and 0.05 h-1, at the beginning of the production phase, are necessary to attain a high P(3HB productivity.

Hollow fibers made from a poly(3-hydroxybutyrate/poly-ε-caprolactone blend

Directory of Open Access Journals (Sweden)

2011-07-01

Full Text Available Since poly(3-hydroxybutyrate (PHB is inherently brittle and possesses poor elastic properties, hollow fibers produced by melt spinning from pure PHB, as described in our earlier study [Macromolecular Materials and Engineering, 2010, 295/6, 585–594], do not meet the required needs regarding the mechanical performance. Besides hardly available PHB copolymers, also blend systems are known to enhance material properties and have thus been considered to be eligible to fabricate flexible or rather pliable hollow fibers based on PHB. Blends of PHB and poly-!-caprolactone (PCL are promising for the application in tissue engineering due to the inherent biocompatibility and biodegradability. A wide range of PHB/PCL compositions have been prepared by melt extrusion. Thermal and mechanical properties of the obtained specimens were analyzed in order to identify miscibility and degree of dispersion as well as to determine the influence on the overall mechanical performance. Even though these constituents are known to be immiscible, PHB/PCL 70/30 was proven to be an adequate composition. This blend showed a highly increased elongation and was found to be easily processable by melt spinning compared to pure PHB. From this blend well defined dimensionally stable bendable hollow fibers were fabricated.

Poly(3-hydroxybutyrate) fuels the tricarboxylic acid cycle and de novo lipid biosynthesis during Bacillus anthracis sporulation.

Science.gov (United States)

Sadykov, Marat R; Ahn, Jong-Sam; Widhelm, Todd J; Eckrich, Valerie M; Endres, Jennifer L; Driks, Adam; Rutkowski, Gregory E; Wingerd, Kevin L; Bayles, Kenneth W

2017-06-01

Numerous bacteria accumulate poly(3-hydroxybutyrate) (PHB) as an intracellular reservoir of carbon and energy in response to imbalanced nutritional conditions. In Bacillus spp., where PHB biosynthesis precedes the formation of the dormant cell type called the spore (sporulation), the direct link between PHB accumulation and efficiency of sporulation was observed in multiple studies. Although the idea of PHB as an intracellular carbon and energy source fueling sporulation was proposed several decades ago, the mechanisms underlying PHB contribution to sporulation have not been defined. Here, we demonstrate that PHB deficiency impairs Bacillus anthracis sporulation through diminishing the energy status of the cells and by reducing carbon flux into the tricarboxylic acid (TCA) cycle and de novo lipid biosynthesis. Consequently, this metabolic imbalance decreased biosynthesis of the critical components required for spore integrity and resistance, such as dipicolinic acid (DPA) and the spore's inner membrane. Supplementation of the PHB deficient mutant with exogenous fatty acids overcame these sporulation defects, highlighting the importance of the TCA cycle and lipid biosynthesis during sporulation. Combined, the results of this work reveal the molecular mechanisms of PHB contribution to B. anthracis sporulation and provide valuable insight into the metabolic requirements for this developmental process in Bacillus species. © 2017 John Wiley & Sons Ltd.

Chloroform induces outstanding crystallization of poly(hydroxybutyrate) (PHB) vesicles within bacteria.

Science.gov (United States)

Rebois, Rolando; Onidas, Delphine; Marcott, Curtis; Noda, Isao; Dazzi, Alexandre

2017-03-01

Poly[(R)-3-hydroxyalkanoate]s or PHAs are aliphatic polyesters produced by numerous microorganisms. They are accumulated as energy and carbon reserve in the form of small intracellular vesicles. Poly[(R)-3-hydroxybutyrate] (PHB) is the most ubiquitous and simplest PHA. An atomic force microscope coupled with a tunable infrared laser (AFM-IR) was used to record highly spatially resolved infrared spectra of commercial purified PHB and native PHB within bacteria. For the first time, the crystallinity degree of native PHB within vesicle has been directly evaluated in situ without alteration due to the measure or extraction and purification steps of the polymer: native PHB is in crystalline state at 15% whereas crystallinity degree reaches 57% in commercial PHB. Chloroform addition on native PHB induces crystallization of the polymer within bacteria up to 60%. This possibility of probing and changing the physical state of polymer in situ could open alternative ways of production for PHB and others biopolymers. Graphical abstract An atomic force microscope coupled with a tunable infrared laser (AFM-IR) has been used to record local infrared spectra of biopolymer PHB within bacteria. Deconvolution of those spectra has allowed to determine in situ the crystallinity degree of native PHB.

The Role of Short-Chain Conjugated Poly-(R-3-Hydroxybutyrate (cPHB in Protein Folding

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Rosetta N. Reusch

2013-05-01

Full Text Available Poly-(R-3-hydroxybutyrate (PHB, a linear polymer of R-3-hydroxybutyrate (R-3HB, is a fundamental constituent of biological cells. Certain prokaryotes accumulate PHB of very high molecular weight (10,000 to >1,000,000 residues, which is segregated within granular deposits in the cytoplasm; however, all prokaryotes and all eukaryotes synthesize PHB of medium-chain length (~100–200 residues which resides within lipid bilayers or lipid vesicles, and PHB of short-chain length (<12 residues which is conjugated to proteins (cPHB, primarily proteins in membranes and organelles. The physical properties of cPHB indicate it plays important roles in the targeting and folding of cPHB-proteins. Here we review the occurrence, physical properties and molecular characteristics of cPHB, and discuss its influence on the folding and structure of outer membrane protein A (OmpA of Escherichia coli.

Characterization of poly(3-hydroxybutyrate) mats obtained by electro spinning method

International Nuclear Information System (INIS)

Goncalves, Joel; Branciforti, Marcia C.; Bretas, Rosario E.S.; Agnelli, Jose A.M.

2009-01-01

Nanofibers of poly-3-hydroxybutyrate (PHB) were produced by electro spinning of chloroform/dimethylformamide (90 / 10% v/v) solutions. The influence of the solution concentration on the processing, morphology and properties of the nanofibers was evaluated. The nanofibers were characterized by wide-angle X-ray diffraction. The nanofibers morphology was evaluated by scanning electron microscopy (SEM). The thermal properties of the nanofibers were analyzed by differential scanning calorimetry (DSC). The results show that the obtained PHB nanofibers exhibit small average diameter (<500 nm) and narrow diameter distribution. A decrease in the fibers average diameters of PHB fibers with the decrease of the solution concentration was observed. The nanofibers were observed to be less crystalline than pure polymer. (author)

Phase diagrams in blends of poly(3-hydroxybutyric acid with various aliphatic polyesters

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2011-07-01

Full Text Available Phase behavior with immiscibility, miscibility, crystalline morphology, and kinetic analysis in blends of poly(3-hydroxybutyric acid (PHB with aliphatic polyesters such as poly(butylene adipate (PBA, poly(ethylene adipate (PEA, poly(trimethylene adipate (PTA, or poly(ethylene succinate (PESu, respectively, were explored mainly using differential scanning calorimeter (DSC and polarized-light optical microscopy (POM. Immiscibility phase behavior with reversible upper-critical-solution-temperature (UCST is common in the PHB/polyester blends. The polyester/polyester blend of PHB/PTA is partially miscible with no UCST in melt and amorphous glassy states within a composition range of PTA less than 50 wt%. The miscible crystalline/crystalline blend exhibits ring-banded spherulites at Tc = 50~100°C, with inter-ring spacing dependent on Tc. All immiscible or partially miscible PHB/polyester blends, by contrast, exhibit disrupted ringbanded spherulites or discrete spherical phase domains upon cooling from UCST to crystallization. The blends of PHB with all other aliphatic polyesters, such as PESu, PEA, PBA, etc. are only partially miscible or immiscible with an upper critical solution temperature (UCST at 180~221°C depending on blend composition. UCST with reversibility was verified.

Biotechnological strategies to improve production of microbial poly-(3-hydroxybutyrate): a review of recent research work

OpenAIRE

Peña, C; Castillo, T; García, A; Millán, M; Segura, D

2014-01-01

Poly-(3-hydroxybutyrate) [P(3HB)] is a polyester synthesized as a carbon and energy reserve material by a wide number of bacteria. This polymer is characterized by its thermo-plastic properties similar to plastics derived from petrochemical industry, such as polyethylene and polypropylene. Furthermore, P(3HB) is an inert, biocompatible and biodegradable material which has been proposed for several uses in medical and biomedical areas. Currently, only few bacterial species such as Cupriavidus ...

Genome-engineered Sinorhizobium meliloti for the production of poly(lactic-co-3-hydroxybutyric) acid copolymer.

Science.gov (United States)

Tran, Tam T; Charles, Trevor C

2016-02-01

Economically competitive commercial production of biodegradable bioplastics with desirable properties is an important goal. In this study, we demonstrate the use of chromosome engineering of an alternative bacterial host, Sinorhizobium meliloti, for production of the copolymer, poly(lactate-co-3-hydroxybutyrate). Codon-optimized genes for 2 previously engineered enzymes, Clostridium propionicum propionate CoA transferase (Pct532Cp) and Pseudomonas sp. strain MBEL 6-19 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1400Ps6-19), were introduced into S. meliloti Rm1021 by chromosome integration, replacing the native phbC gene. On the basis of phenotypic analysis and detection of polymer product by gas chromatography analysis, synthesis and accumulation of the copolymer was confirmed. The chromosome integrant strain, with the introduced genes under the control of the native phbC promoter, is able to produce over 15% cell dry mass of poly(lactate-co-3-hydroxybutyrate), containing 30 mol% lactate, from growth on mannitol. We were also able to purify the polymer from the culture and confirm the structure by NMR and GC-MS. To our knowledge, this is the first demonstration of production of this copolymer in the Alphaproteobacteria. Further optimization of this system may eventually yield strains that are able to produce economically viable commercial product.

Thermal stability of poly(3-hydroxybutyrate)/vegetable fiber composites

Science.gov (United States)

Cipriano, Pâmela Bento; de Sá, Mayelli Dantas; Andrade, André L. Simões; de Carvalho, Laura Hecker; Canedo, Eduardo Luis

2015-05-01

The present work deals with the thermal stability during and after processing of composites of poly(3-hydroxybutyrate) (PHB) - a fully biodegradable semi-crystalline thermoplastic obtained from renewable resources through low-impact biotechnological process, biocompatible and non-toxic - and vegetable fiber from the fruit (coconut) of babassu palm tree. PHB/babassu composites with 0, 5, 10 and 20% w/w load were prepared in a laboratory internal mixer. Two fractions of the mesocarp of babassu with different particle sizes were compounded with PHB and test specimens molded by compression. The effect of loading level and processing conditions on torque, temperature and mechanical energy dissipation were studied using a new engineering model. It was found that PHB degrades during processing at temperatures slightly above the melting point. To minimize thermal degradation stabilizer and chain extender additives were incorporated, with mixed results. These findings were confirmed by the dependence of the melt flow rate on the processing temperature.

Targeted poly(3-hydroxybutyrate-co-3-hydroxyvalerate) bioplastic production from carbon dioxide.

Science.gov (United States)

Ghysels, Stef; Mozumder, Md Salatul Islam; De Wever, Heleen; Volcke, Eveline I P; Garcia-Gonzalez, Linsey

2018-02-01

A microbial production process was developed to convert CO 2 and valeric acid into tailored poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) bioplastics. The aim was to understand microbial PHBV production in mixotrophic conditions and to control the monomer distribution in the polymer. Continuous sparging of CO 2 with pulse and pH-stat feeding of valeric acid were evaluated to produce PHBV copolyesters with predefined properties. The desired random monomer distribution was obtained by limiting the valeric acid concentration (below 1 gL -1 ). 1 H-NMR, 13 C-NMR and chromatographic analysis of the PHBV copolymer confirmed both the monomer distribution and the 3-hydroxyvalerate (3HV) fraction in the produced PHBV. A physical-based model was developed for mixotrophic PHBV production, which was calibrated and validated with independent experimental datasets. To produce PHBV with a predefined 3HV fraction, an operating diagram was constructed. This tool was able to predict the 3HV fraction with a very good accuracy (2% deviation). Copyright © 2017 Elsevier Ltd. All rights reserved.

Dexamethasone-loaded poly(3-hydroxybutyrate-co-3-hydroxyvalerate) microparticles for controlled release

International Nuclear Information System (INIS)

Riekes, Manoela Klueppel; Paula, Josiane Padilha de; Farago, Paulo Vitor; Zawadzki, Sonia Faria

2009-01-01

Dexamethasone (DEX) has been widely used for the treatment of ulcerative colitis. The aim of the present study was to obtain DEX-loaded poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) microparticles prepared by simple emulsion/solvent evaporation method. The drug loading and the encapsulation efficiency were determined by a previously validated UV method at 233 nm. Morphological, spectroscopical and dissolution analyses were also performed. The microparticles (formulation F no. 0, F no. 1 and F no. 2) were successfully obtained as off-white powders. A drug loading of 92.27 mg.g -1 and 218.54 mg.g -1 and an encapsulation efficiency of 93.96 % and 87.43 % were respectively observed for F no. 1 and F no. 2. Particles showed spherical and rough aspect by SEM. X-ray diffraction analysis demonstrated that the encapsulation reduced the drug crystallinity. FTIR spectra showed that no chemical bonding occurred between PHBV and DEX. Drug-loaded microparticles revealed controlled release profiles compared to pure DEX. (author)

Water and vapor permeability at different temperatures of poly (3-Hydroxybutyrate dense membranes

Directory of Open Access Journals (Sweden)

Luiz H. Poley

2005-03-01

Full Text Available Polyhydroxyalkanoates (PHAs are polymers produced from renewable resources with biodegradability and biocompatibility, being therefore attractive for medical and pharmaceutical purposes. Poly (3-hydroxybutyrate (PHB is the most important polymer of this family by considering the biotechnology process of its synthesis. In the present study, dense films of PHB were prepared by casting from chloroform solutions (1% m/m. Permeability studies with water, methanol, ethanol and n-propanol were performed using the gravimetric method at different temperatures (from 50 ºC to 65 ºC. Results provide new data on permeability coefficients of PHB membranes.

Preparation of poly(3-hydroxybutyrate)/carboxymethyl cellulose acetate butyrate blends using gel formation

International Nuclear Information System (INIS)

Gomes, A.L.; Rodrigues, G.V.; Goncalves, M.C.

2009-01-01

This study investigates poly(3-hydroxybutyrate) (PHB) gel formation with a binary combination of solvents and its use on the preparation of PHB and carboxymethyl cellulose acetate butyrate (CMCAB) blends. The gel preparation method was compared to a precipitation method followed by hot pressing. The results from DSC and X-ray diffractions showed that both methodologies produced blends with very similar thermal properties and crystallization behavior. Scanning electron microscopy indicated better homogeneity in gel formation blends. Apart from this, the gel formation methodology provided new ways to prepare immiscible blends with the advantage of using friendlier solvents. (author)

Dual production of poly(3-hydroxybutyrate) and glutamate using variable biotin concentrations in Corynebacterium glutamicum.

Science.gov (United States)

Jo, Sung-Jin; Leong, Chean Ring; Matsumoto, Ken'ichiro; Taguchi, Seiichi

2009-04-01

We previously synthesized poly(3-hydroxybutyrate) [P(3HB)] in recombinant Corynebacterium glutamicum, a prominent producer of amino acids. In this study, a two-step cultivation was established for the dual production of glutamate and P(3HB) due to the differences in the optimal concentration of biotin. Glutamate was extracellularly produced first under the biotin-limited condition of 0.3 microg/L. Production was then shifted to P(3HB) by addition of biotin to a total concentration of 9 microg/L. The final products obtained were 18 g/L glutamate and 36 wt% of P(3HB).

Thermal, mechanical and morphological properties of poly (hydroxybutyrate and polypropylene blends after processing

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Wagner Mauricio Pachekoski

2009-06-01

Full Text Available The ever increasing accumulation of plastic waste in the environment has motivated research on polymers that degrade rapidly after being discarded as possible substitutes for conventional inert plastics. Biodegradable polymers can be an alternative, since they have non-toxic residual products and low environmental permanence. Poly (hydroxybutyrate is a biodegradable polymer with a strong potential for industrial purposes, but its thermal instability and fragility limit its applications. Thus, an alternative to improve the processability and properties of poly (hydroxybutyrate is to mix it with another polymer, not necessarily a biodegradable one. In this work, different mixtures of poly(hydroxybutyrate or PHB and polypropylene or PP were extruded and injected. After processing, the blends were studied and their miscibility, mechanical properties and degradability in different soils were analyzed. The main results indicated that the PHB/PP blends had better mechanical properties than pure PHB, as well as improved immiscibility and higher degradation in alkaline soil. The poly-hydroxybutyrate/polypropylene blends showed a tendency for lower crystallinity and stiffness of the polymer matrix, proportional to the amount of polypropylene in the blends, rendering them less stiff and fragile. The degradation tests showed that both pure PHB and blends with 90% PHB and 10% PP were degraded, with loss of their mechanical properties and weight.

Poly-(R)-3-hydroxybutyrates (PHB) are Atherogenic Components of Lipoprotein Lp(a).

Science.gov (United States)

Reusch, Rosetta N

2015-12-01

The hypothesis is that poly-(R)-3-hydroxybutyrates (PHB), linear polymers of the ketone body, R-3-hydroxybutyrate (R-3HB), are atherogenic components of lipoprotein Lp(a). PHB are universal constituents of biological cells and are thus components of all foods. Medium chain-length PHB (PHB (PHB are highly insoluble in water, but soluble in lipids in which they exhibit a high intrinsic viscosity. They have a higher density than other cellular lipids and they are very adhesive, i.e. they engage in multiple noncovalent interactions with other molecules and salts via hydrogen, hydrophobic and coordinate bonds, thus producing insoluble deposits. Following digestive processes, PHB enter the circulation in chylomicrons and very low density lipoproteins (VLDL). The majority of the PHB (>70%) are absorbed by albumin, which transports them to the liver for disposal. When the amount of PHB in the diet exceed the capacity of albumin to safely remove them from the circulation, the excess PHB remain in the lipid core of LDL particles that become constituents of lipoprotein Lp(a), and contribute to the formation of arterial deposits. In summary, the presence of PHB – water-insoluble, dense, viscous, adhesive polymers – in the lipid cores of the LDL moieties of Lp(a) particles supports the hypothesis that PHB are atherogenic components of Lp(a).

Synthesis, characterizations and biocompatibility of novel biodegradable star block copolymers based on poly[(R)-3-hydroxybutyrate] and poly(epsilon-caprolactone)

DEFF Research Database (Denmark)

Wu, Linping; Wang, Liang; Wang, Xiaojuan

2010-01-01

Star block copolymers based on poly[(R)-3-hydroxybutyrate] (PHB) and poly(epsilon-caprolactone) (PCL), termed SPHBCL, were successfully synthesized with structural variation on arm numbers and lengths via coupling reactions and ring opening polymerizations. Arm numbers 3, 4 and 6 of SPHBCL were...... weights of the SPHBCL due to the discrepancy of star copolymer structures. The melting temperature of SPHBCL decreased with increasing degree of branching. Thermal decomposition temperature was revealed to be lower than that of linear block copolymer LPHBCL counterparts based on PHB and PCL. Films made...... from various SPHBCL copolymers had different porous or networking surface morphology, and all possessed improved biocompatibility in terms of less blood clotting and more osteoblast cell growth compared with their corresponding homopolymers PHB and PCL. Among them, it was found, however, that the 4-arm...

Role of β-hydroxybutyrate, its polymer poly-β-hydroxybutyrate and inorganic polyphosphate in mammalian health and disease

Directory of Open Access Journals (Sweden)

Elena N. Dedkova

2014-07-01

Full Text Available We provide a comprehensive review of the role of β-hydroxybutyrate (β-OHB, its linear polymer poly-β-hydroxybutyrate (PHB and inorganic polyphosphate (polyP in mammalian health and disease. β-OHB is a metabolic intermediate that constitutes 70% of ketone bodies produced during ketosis. Although ketosis has been generally considered as an unfavorable pathological state (e.g. diabetic ketoacidosis in type-1 diabetes mellitus, it has been suggested that induction of mild hyperketonemia may have certain therapeutic benefits. β-OHB is synthesized in the liver from acetyl-CoA by β-hydroxybutyrate dehydrogenase and can be used as alternative energy source. Elevated levels of PHB are associated with pathological states. In humans, short-chain, complexed PHB (cPHB is found in a wide variety of tissues and in atherosclerotic plaques. Plasma cPHB concentrations correlate strongly with atherogenic lipid profiles, and PHB tissue levels are elevated in type-1 diabetic animals. However, little is known about mechanisms of PHB action especially in the heart. In contrast to β-OHB, PHB is a water-insoluble, amphiphilic polymer that has high intrinsic viscosity and salt-solvating properties. cPHB can form non-specific ion channels in planar lipid bilayers and liposomes. PHB can form complexes with polyP and Ca2+ which increases membrane permeability. The biological roles played by polyP, a ubiquitous phosphate polymer with ATP-like bonds, have been most extensively studied in prokaryotes, however polyP has recently been linked to a variety of functions in mammalian cells, including blood coagulation, regulation of enzyme activity in cancer cells, cell proliferation, apoptosis and mitochondrial ion transport and energy metabolism. Recent evidence suggests that polyP is a potent activator of the mitochondrial permeability transition pore in cardiomyocytes and may represent a hitherto unrecognized key structural and functional component of the mitochondrial

Enhanced poly(3-hydroxybutyrate) production in transgenic tobacco BY-2 cells using engineered acetoacetyl-CoA reductase.

Science.gov (United States)

Yokoo, Toshinori; Matsumoto, Ken'ichiro; Ooba, Takashi; Morimoto, Kenjiro; Taguchi, Seiichi

2015-01-01

Highly active mutant of NADPH-dependent acetoacetyl-CoA reductase (PhaB) was expressed in Nicotiana tabacum cv. Bright Yellow-2 cultured cells to produce poly(3-hydroxybutyrate) [P(3HB)]. The mutated PhaB increased P(3HB) content by three-fold over the control, indicating that the mutant was a versatile tool for P(3HB) production. Additionally, the PhaB-catalyzed reaction was suggested to be a rate-limiting step of P(3HB) biosynthesis in tobacco BY-2 cells.

Functional Properties of Plasticized Bio-Based Poly(Lactic Acid)_Poly(Hydroxybutyrate) (PLA_PHB) Films for Active Food Packaging

OpenAIRE

Burgos, Nuria; Armentano, Ilaria; Fortunati, Elena; Dominici, Franco; Luzi, Francesca; Fiori, Stefano; Cristofaro, Francesco; Visai, Livia; Jiménez, Alfonso; Kenny, José María

2017-01-01

Fully bio-based and biodegradable active films based on poly(lactic acid) (PLA) blended with poly(3-hydroxybutyrate) (PHB) and incorporating lactic acid oligomers (OLA) as plasticizers and carvacrol as active agent were extruded and fully characterized in their functional properties for antimicrobial active packaging. PLA_PHB films showed good barrier to water vapor, while the resistance to oxygen diffusion decreased with the addition of OLA and carvacrol. Their overall migration in aqueous f...

Nanofiber-bonded cloth materials based on poly-3-hydroxybutyrate with antibacterial properties for medical purposes

Science.gov (United States)

Tyubaeva, P. M.; Olkhov, A. A.; Karpova, S. G.; Iordansky, A. L.; Popov, A. A.

2017-12-01

Different transdermal systems based on solid polymer matrices or gels containing functional substances with antiseptic (antibacterial) properties have application to the therapy of many infectious diseases and cancer. Today the most promising type of matrices with antiseptic characteristics are the nano- and microfiber nonwoven materials. Fibers on the biopolymer (poly(3-hydroxybutyrate)) basis were obtained using the electrospinning method. In the present work, the effects of iron (III) complex with tetraphenylporphyrin and its influence on bactericidal and antibacterial properties of the ultrathin PHB fibers were investigated.

Microcellular poly(hydroxybutyrate-co-hydroxyvalerate)-hyperbranched polymer-nanoclay nanocomposites

Science.gov (United States)

Alireza Javadi; Yottha Srithep; Srikanth Pilla; Craig C. Clemons; Shaoqin Gong; Lih-Sheng Turng

2012-01-01

The effects of incorporating hyperbranched polymers (HBPs) and different nanoclays [Cloisite® 30B and halloysite nanotubes (HNT)] on the mechanical, morphological, and thermal properties of solid and microcellular poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) were investigated. According to the X-ray diffraction (...

Effect of Graphite Nanosheets on Properties of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate

Directory of Open Access Journals (Sweden)

Larissa Stieven Montagna

2017-01-01

Full Text Available The influence of different contents, 0.25, 0.50, and 1.00 wt%, of graphite nanosheets (GNS on the properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV nanocomposites obtained by solution casting method has been studied. GNS were prepared by three steps: intercalation (chemical exfoliation, expansion (thermal treatment, and the GNS obtainment (physical treatment by ultrasonic exfoliation. X-ray diffraction (XRD, Raman spectroscopy, and field emission gun-scanning electron microscopy (FE-SEM showed that the physical, chemical, and thermal treatments preserved the graphite sheets structure. XRD and Raman results also showed that GNS were dispersed in the PHBV matrix. The degree of crystallinity (Xc of the nanocomposites did not change when the graphite nanosheets were added. However, the GNS acted as nucleation agent for crystallization; that is, in the second heating the samples containing GNS showed two melting peaks. The addition the GNS did not change the thermal stability of the PHBV.

Preparation and characterization of ibuprofen-loaded microspheres consisting of poly(3-hydroxybutyrate) and methoxy poly (ethylene glycol)-b-poly (D,L-lactide) blends or poly(3-hydroxybutyrate) and gelatin composites for controlled drug release

Energy Technology Data Exchange (ETDEWEB)

Bidone, Juliana; Melo, Ana Paula P. [Laboratorio de Farmacotecnica, Departamento de Ciencias Farmaceuticas, Universidade Federal de Santa Catarina, Florianopolis (Brazil); Bazzo, Giovana C. [Grupo de Estudos em Materiais Polimericos (POLIMAT), Departamento de Quimica, Universidade Federal de Santa Catarina, Florianopolis (Brazil); Carmignan, Francoise [Laboratorio de Farmacotecnica, Departamento de Ciencias Farmaceuticas, Universidade Federal de Santa Catarina, Florianopolis (Brazil); Soldi, Marli S.; Pires, Alfredo T.N. [Grupo de Estudos em Materiais Polimericos (POLIMAT), Departamento de Quimica, Universidade Federal de Santa Catarina, Florianopolis (Brazil); Lemos-Senna, Elenara [Laboratorio de Farmacotecnica, Departamento de Ciencias Farmaceuticas, Universidade Federal de Santa Catarina, Florianopolis (Brazil)], E-mail: lemos@ccs.ufsc.br

2009-03-01

Poly-(3-hydroxybutyrate) (P(3HB)) is a biodegradable and biocompatible polymer that has been used to obtain polymer-based drug carriers. However, due to the high crystallinity degree of this polymer, drug release from P(3HB) microspheres frequently occurs at excessive rates. In this study, two strategies for prolonging ibuprofen release from P(3HB)-based microspheres were tested: blending with poly(D,L-lactide)-b-polyethylene glycol (mPEG-PLA); and obtaining composite particles with gelatin (GEL). SEM micrographs showed particles that were spherical and had a rough surface. A slight decrease of the crystallinity degree of P(3HB) was observed only in the DSC thermogram obtained from unloaded-microspheres prepared from 1:1 P(3HB):mPEG-PLA blend. For IBF-loaded microspheres, a reduction of around 10 deg. C in the melting temperature of P(3HB) was observed, indicating that the crystalline structure of the polymer was affected in the presence of the drug. DSC studies also yielded evidence of the presence of a molecular dispersion coexisting with a crystalline dispersion in the drug in the matrix. Similar results were obtained from X-ray diffractograms. In spite of 1:1 mPEG-PLA:P(3HB) blends having contributed to the reduction of the burst effect, a more controlled drug release was provided by the use of the 3:1 P(3HB):mPEGPLA blend. This result indicated that particle hydration played an important role in the drug release. On the other hand, the preparation of P(3HB):GEL composite microspheres did not allow control of the IBF release.

Preparation and characterization of ibuprofen-loaded microspheres consisting of poly(3-hydroxybutyrate) and methoxy poly (ethylene glycol)-b-poly (D,L-lactide) blends or poly(3-hydroxybutyrate) and gelatin composites for controlled drug release

International Nuclear Information System (INIS)

Bidone, Juliana; Melo, Ana Paula P.; Bazzo, Giovana C.; Carmignan, Francoise; Soldi, Marli S.; Pires, Alfredo T.N.; Lemos-Senna, Elenara

2009-01-01

Poly-(3-hydroxybutyrate) (P(3HB)) is a biodegradable and biocompatible polymer that has been used to obtain polymer-based drug carriers. However, due to the high crystallinity degree of this polymer, drug release from P(3HB) microspheres frequently occurs at excessive rates. In this study, two strategies for prolonging ibuprofen release from P(3HB)-based microspheres were tested: blending with poly(D,L-lactide)-b-polyethylene glycol (mPEG-PLA); and obtaining composite particles with gelatin (GEL). SEM micrographs showed particles that were spherical and had a rough surface. A slight decrease of the crystallinity degree of P(3HB) was observed only in the DSC thermogram obtained from unloaded-microspheres prepared from 1:1 P(3HB):mPEG-PLA blend. For IBF-loaded microspheres, a reduction of around 10 deg. C in the melting temperature of P(3HB) was observed, indicating that the crystalline structure of the polymer was affected in the presence of the drug. DSC studies also yielded evidence of the presence of a molecular dispersion coexisting with a crystalline dispersion in the drug in the matrix. Similar results were obtained from X-ray diffractograms. In spite of 1:1 mPEG-PLA:P(3HB) blends having contributed to the reduction of the burst effect, a more controlled drug release was provided by the use of the 3:1 P(3HB):mPEGPLA blend. This result indicated that particle hydration played an important role in the drug release. On the other hand, the preparation of P(3HB):GEL composite microspheres did not allow control of the IBF release

Mixed Cultures Fermentation for the Production of Poly-ß-hydroxybutyrate

Directory of Open Access Journals (Sweden)

Thomas Shalin

2014-10-01

Full Text Available Poly-ß-hydroxybutyrate (PHB is a biodegradable intracellular microbial product produced by many bacteria and it is comparable to some of the petrochemical derived thermoplastics such as polypropylene. One of the main barriers for the commercial exploitation is the high cost of the substrate for the production of biopolymer. The utilization of mixed microbial cultures facilitates the use of complex substrates thereby reducing the cost of PHB production. In the present study, mixed culture systems were evaluated for PHB production. Bacillus firmus NII 0830 was used for the production of PHB since it accumulates a large amount of PHB and a second organism Lactobacillus delbrueckii NII 0925 was used to provide lactic acid. FTIR and 1H NMR analyses revealed that the PHB extracted from pure culture and mixed culture showed exact match to that of standard PHB. Biodegradation studies of the PHB blends showed 87% degradation. It was also found that a consortium of organisms degraded the films faster than a single organism.

Composite scaffolds for cartilage tissue engineering based on natural polymers of bacterial origin, thermoplastic poly(3-hydroxybutyrate) and micro-fibrillated bacterial cellulose

Czech Academy of Sciences Publication Activity Database

Akaraonye, E.; Filip, J.; Šafaříková, Miroslava; Salih, V.; Keshavarz, T.; Knowles, J.C.; Roy, I.

2016-01-01

Roč. 65, č. 7 (2016), s. 780-791 ISSN 0959-8103 Institutional support: RVO:60077344 Keywords : polyhydroxyalkanoates * poly(3-hydroxybutyrate) * bacterial cellulose * micro-fibrillated cellulose * tissue engineering scaffold * composite materials Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.070, year: 2016

Melting and thermal history of poly(hydroxybutyrate-co-hydroxyvalerate) using step-scan DSC

International Nuclear Information System (INIS)

Gunaratne, L.M.W.K.; Shanks, R.A.

2005-01-01

Melting behaviour and crystal morphology of poly(3-hydroxybutyrate) (PHB) and its copolymer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with various hydroxyvalerate (HV) contents [5 wt.% (PHB5HV), 8 wt.% (PHB8HV) and 12 wt.% (PHB12HV)] have been investigated by conventional DSC, step-scan differential scanning calorimetry (SDSC) and hot-stage polarised optical microscopy (HSPOM). Crystallisation behaviour of PHB and its copolymers were investigated by SDSC. Thermal properties were investigated after different crystallisation treatments, fast, medium and slow cooling. Multiple melting peak behaviour was observed for all polymers. SDSC data revealed that PHB and its copolymers undergo melting-recrystallisation-remelting during heating, as evidenced by exothermic peaks in the IsoK baseline (non-reversing signal). An increase in degree of crystallinity due to significant melt-recrystallisation was observed for slow-cooled copolymers. PHB5HV showed different crystal morphologies for various crystallisation conditions. SDSC proved a convenient and precise method for measurement of the apparent thermodynamic specific heat (reversing signal) HSPOM results showed that the crystallisation rates and sizes of spherulites were significantly reduced as crystallisation rate increased

Effects of solvents on the radiation grafting reaction of vinyl compounds on poly (3-hydroxybutyrate)

International Nuclear Information System (INIS)

Torres, Maykel González; Talavera, José Rogelio Rodríguez; Muñoz, Susana Vargas; Pérez, Manuel González; Castro, Ma. Pilar. Carreón.; Cortes, Jorge Cerna

2015-01-01

Vinyl Acetate was grafted onto poly (3-hydroxybutyrate) by the simultaneous gamma irradiation method using different types of solvents and in bulk (solvent free), at 10 kGy and 1.62 kGy/h dose and dose rate respectively. Subsequent complete hydrolysis allowed the conversion of grafted chains from poly (vinyl acetate) to poly (vinyl alcohol). The aim of this study is to determine the effect of solvent through the estimation of the dependence of the degree of grafting with the choice of solvent, the calculation of the degree of crystallinity, and to study the biodegradation of the products. The results showed a greater degree of grafting in bulk, while the more suitable solvent was hexane. Characterization of the grafted copolymer indicated that crystallinity percentage decreased by an increase in grafting, while the biodegradability was promoted by the increment in poly (vinyl alcohol) grafted. - Highlights: • PHB was indirectly grafted with PVA, by complete hydrolysis of grafted PVAc. • The effect of solvents on the grafting, crystallinity and biodegradation was studied. • The characterizations of the products were obtained by SEM, TGA, and DSC

Glycolysis of poly(3-hydroxybutyrate) via enzyme catalysis

International Nuclear Information System (INIS)

Paula, Everton Luiz de; Campos, Tiago Ferreira; Mano, Valdir

2014-01-01

Poly(3-hydroxybutyrate), PHB, is a polymer with broad potential applications because of its biodegradability and biocompatibility. However, its high crystallinity is a limiting factor for many applications. To overcome this drawback, one strategy currently employed involves the reduction of the molecular weight of PHB with the concomitant formation of end-functionalized chains, such as those obtained via glycolysis. The glycolysis of PHB can be catalyzed by acid, base, or organometallic compounds. However, to our knowledge, there are no reports regarding PHB glycolysis catalyzed enzymatically. Among the major types of enzymes used in biocatalysis, the lipases stand out because they have the ability to catalyze reactions in both aqueous and organic media. Thus, in this study, we performed the enzymatic glycolysis of PHB using the lipase Amano PS (Pseudomonas cepacia) with ethane-1,2-diol (ethylene glycol) as the functionalizing agent. The results indicated that the glycolysis was successful and afforded hydroxyl-terminated oligomeric PHB polyols. Nuclear magnetic resonance spectra of the products showed characteristic signals for the terminal hydroxyl groups of the polyols, while thermogravimetric and differential scanning calorimetry analyses confirmed an increase in the thermal stability and a decrease in the crystallinity of the polyols compared with the starting PHB polymer, which were both attributed to the reduction in the molecular weight due to glycolysis. (author)

Draft Genome Sequence of Bacillus aryabhattai Strain PHB10, a Poly(3-Hydroxybutyrate)-Accumulating Bacterium Isolated from Domestic Sewerage.

Science.gov (United States)

Balakrishna Pillai, Aneesh; Jaya Kumar, Arjun; Thulasi, Kavitha; Reghunathan, Dinesh; Prasannakumar, Manoj; Kumarapillai, Harikrishnan

2017-10-12

Bacillus aryabhattai PHB10 is a poly(3-hydroxybutyrate) (PHB)-accumulating bacterium isolated from domestic sewerage. Here, we report the 4.19-Mb draft genome sequence, with 4,050 protein-coding genes and a G+C content of 37.5%. This sequence will be helpful in the study of the high-level PHB accumulation mechanism of the strain. Copyright © 2017 Balakrishna Pillai et al.

Chemical synthesis of perfectly isotactic and high melting bacterial poly(3-hydroxybutyrate) from bio-sourced racemic cyclic diolide.

Science.gov (United States)

Tang, Xiaoyan; Chen, Eugene Y-X

2018-06-11

Bacterial poly(3-hydroxybutyrate) (P3HB) is a perfectly isotactic, crystalline material possessing properties suitable for substituting petroleum plastics, but high costs and low volumes of its production are impractical for commodity applications. The chemical synthesis of P3HB via ring-opening polymerization (ROP) of racemic β-butyrolactone has attracted intensive efforts since the 1960s, but not yet produced P3HB with high isotacticity and molecular weight. Here, we report a route utilizing racemic cyclic diolide (rac-DL) derived from bio-sourced succinate. With stereoselective racemic catalysts, the ROP of rac-DL under ambient conditions produces rapidly P3HB with perfect isotacticity ([mm] > 99%), high melting temperature (T m  = 171 °C), and high molecular weight (M n  = 1.54 × 10 5  g mol -1 , Đ = 1.01). With enantiomeric catalysts, kinetic resolution polymerizations of rac-DL automatically stops at 50% conversion and yields enantiopure (R,R)-DL and (S,S)-DL with >99% e.e. and the corresponding poly[(S)-3HB] and poly[(R)-3HB] with high T m  = 175 °C.

Analysis of structure of hyperfine poly(3-hydroxybutyrate) fibers (PHB) for controlled drug delivery

Science.gov (United States)

Olkhov, A. A.; Kosenko, R. Yu; Markin, V. S.; Zykova, A. K.; Pantyukhov, P. V.; Karpova, S. G.; Iordanskii, A. L.

2017-12-01

Hyperfine fibers based on biodegradable poly (3-hydroxybutyrate) with encapsulated drug substance (dipyridamol) were obtained by using electrospinning method. Addition of dipyridamol has a significant effect on geometrical shape and structure of microfibers as well as total porosity of fibrous material. Observation of fibers using scanning electron microscopy (SEM) method showed that without or at lower dipyridamol content (<3%) fibers consisted of interleaved ellipsoid and cylindrical fragments. At higher dipyridamol content (3-5%) anomalous ellipsoid structures did not practically form, and fiber’s shape became cylindrical. The totality of morphological and structural characteristics determined the rate of dipyridamol diffusive transports. The simplified model of drug desorption from fibrous matrix was presented. In current work it was showed that the rate-limiting stage of transport was the diffusion of dipyridamol in the bulk of cylindrical fibers.

Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant Escherichia coli

DEFF Research Database (Denmark)

Zhou, Xiao-Yun; Yuan, Xiao-Xi; Shi, Zhen-Yu

2012-01-01

inactivated to enhance the carbon flux to poly(4HB) biosynthesis. Four PHA binding proteins (PhaP or phasins) including PhaP1, PhaP2, PhaP3 and PhaP4 from R. eutropha were heterologously expressed in the recombinant E. coli, respectively, leading to different levels of improvement in poly(4HB) production......-hydroxybutyrate or 1,4-butanediol (1,4-BD) are provided as precursor which are much more expensive than glucose. At present, high production cost is a big obstacle for large scale production of poly(4HB). RESULTS: Recombinant Escherichia coli strain was constructed to achieve hyperproduction of poly(4....... Among them PhaP1 exhibited the highest capability for enhanced polymer synthesis. The recombinant E. coli produced 5.5 g L(-1) cell dry weight containing 35.4% poly(4HB) using glucose as a sole carbon source in a 48 h shake flask growth. In a 6-L fermentor study, 11.5 g L(-1) cell dry weight containing...

Influence of gamma radiation on thermal properties and water vapor transmission of poly(3-hydroxybutyrate) (PHB) in blends

Energy Technology Data Exchange (ETDEWEB)

Forster, Pedro L.; Martins, Natalia A.; Parra, Duclerc F.; Egute, Nayara S.; Lugao, Ademar B., E-mail: dfparra@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

2009-07-01

Biodegradable polymers are a newly emerging field. A vast number of biodegradable polymers have been synthesized recently and some microorganisms and enzymes capable of degrading them have been identified. Polyesters such as poly(3-hydroxybutyrate) (PHB) or other polyhydroxyalkanoates (PHAs) have attracted commercial and academic interest as new biodegradable materials. In this work, we investigated the effect of gamma radiation on the thermal properties and biodegradation behavior of PHB in blend with poly(ethyleneglycol)(PEG). The samples were irradiated at gamma radiation of 5 and 10 kGy. The thermal behaviour was investigated by utilization of differential scanning calorimetry (DSC) changes in thermal stability, glass transition and melting point were reported. (author)

Influence of gamma radiation on thermal properties and water vapor transmission of poly(3-hydroxybutyrate) (PHB) in blends

International Nuclear Information System (INIS)

Forster, Pedro L.; Martins, Natalia A.; Parra, Duclerc F.; Egute, Nayara S.; Lugao, Ademar B.

2009-01-01

Biodegradable polymers are a newly emerging field. A vast number of biodegradable polymers have been synthesized recently and some microorganisms and enzymes capable of degrading them have been identified. Polyesters such as poly(3-hydroxybutyrate) (PHB) or other polyhydroxyalkanoates (PHAs) have attracted commercial and academic interest as new biodegradable materials. In this work, we investigated the effect of gamma radiation on the thermal properties and biodegradation behavior of PHB in blend with poly(ethyleneglycol)(PEG). The samples were irradiated at gamma radiation of 5 and 10 kGy. The thermal behaviour was investigated by utilization of differential scanning calorimetry (DSC) changes in thermal stability, glass transition and melting point were reported. (author)

Biodegradability of poly(3-hydroxybutyrate) film grafted with vinyl acetate: Effect of grafting and saponification

Science.gov (United States)

Wada, Yuki; Seko, Noriaki; Nagasawa, Naotsugu; Tamada, Masao; Kasuya, Ken-ichi; Mitomo, Hiroshi

2007-06-01

Radiation-induced graft polymerization of vinyl acetate (VAc) onto poly(3-hydroxybutyrate) (PHB) film was carried out. At a degree of grafting higher than 5%, the grafted films (PHB-g-VAc) completely lost the enzymatic degradability that is characteristic of PHB due to the grafted VAc covering the surface of the PHB film. However, the biodegradability of the PHB-g-VAc films was recovered when the films were saponified in alkali solution under optimum conditions. Graft chains of the PHB-g-VAc film reacted selectively to become biodegradable polyvinyl alcohol (PVA). The biodegradability of the saponified PHB-g-VAc film increased rapidly with time.

Biodegradability of poly(3-hydroxybutyrate) film grafted with vinyl acetate: Effect of grafting and saponification

International Nuclear Information System (INIS)

Wada, Yuki; Seko, Noriaki; Nagasawa, Naotsugu; Tamada, Masao; Kasuya, Ken-ichi; Mitomo, Hiroshi

2007-01-01

Radiation-induced graft polymerization of vinyl acetate (VAc) onto poly(3-hydroxybutyrate) (PHB) film was carried out. At a degree of grafting higher than 5%, the grafted films (PHB-g-VAc) completely lost the enzymatic degradability that is characteristic of PHB due to the grafted VAc covering the surface of the PHB film. However, the biodegradability of the PHB-g-VAc films was recovered when the films were saponified in alkali solution under optimum conditions. Graft chains of the PHB-g-VAc film reacted selectively to become biodegradable polyvinyl alcohol (PVA). The biodegradability of the saponified PHB-g-VAc film increased rapidly with time

Surface wettability and energy effects on the biological performance of poly-3-hydroxybutyrate films treated with RF plasma

Energy Technology Data Exchange (ETDEWEB)

Syromotina, D.S. [Department of Experimental Physics, National Research Tomsk Polytechnic University, 634050 Tomsk (Russian Federation); Surmenev, R.A., E-mail: rsurmenev@gmail.com [Department of Experimental Physics, National Research Tomsk Polytechnic University, 634050 Tomsk (Russian Federation); Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, 70569 Stuttgart (Germany); Surmeneva, M.A. [Department of Experimental Physics, National Research Tomsk Polytechnic University, 634050 Tomsk (Russian Federation); Boyandin, A.N.; Nikolaeva, E.D. [Institute of Biophysics of Siberian Branch of Russian Academy of Sciences, 50/50 Akademgorodok, Krasnoyarsk 660036 (Russian Federation); School of Fundamental Biology and Biotechnology, Siberian Federal University, 79 Svobodny pr., 660041 Krasnoyarsk (Russian Federation); Prymak, O.; Epple, M. [Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, 45117 Essen (Germany); Ulbricht, M. [Technical Chemistry II and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, 45141 Essen (Germany); Oehr, C. [Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, 70569 Stuttgart (Germany); Volova, T.G. [Institute of Biophysics of Siberian Branch of Russian Academy of Sciences, 50/50 Akademgorodok, Krasnoyarsk 660036 (Russian Federation); School of Fundamental Biology and Biotechnology, Siberian Federal University, 79 Svobodny pr., 660041 Krasnoyarsk (Russian Federation)

2016-05-01

The surface properties of poly-3-hydroxybutyrate (P3HB) membranes were modified using oxygen and an ammonia radio-frequency (RF, 13.56 MHz) plasma. The plasma treatment procedures used in the study only affected the surface properties, including surface topography, without inducing any significant changes in the crystalline structure of the polymer, with the exception being a power level of 250 W. The wettability of the modified P3HB surfaces was significantly increased after the plasma treatment, irrespective of the treatment procedure used. It was revealed that both surface chemistry and surface roughness changes caused by the plasma treatment affected surface wettability. A treatment-induced surface aging effect was observed and resulted in an increase in the water contact angle and a decrease in the surface free energy. However, the difference in the water contact angle between the polymers that had been treated for 4 weeks and the untreated polymer surfaces was still significant. A dependence between cell adhesion and proliferation and the polar component of the surface energy was revealed. The increase in the polar component after the ammonia plasma modification significantly increased cell adhesion and proliferation on biodegradable polymer surfaces compared to the untreated P3HB and the P3HB modified using an oxygen plasma. - Highlights: • Plasma treatment affected the topography of poly(3-hydroxybutyrate) (P3HB). • Plasma treatment resulted in improvement of the surface wettability. • No alteration of the bulk properties of the polymers was observed. • The ammonia plasma treatment at 150 W improved the cell adhesion and proliferation.

Analysis of the structure of poly-3-hydroxybutyrate ultrathin fibers modified with iron (III) complex with tetraphenylporphyrin

Science.gov (United States)

Olkhov, A. A.; Karpova, S. G.; Lobanov, A. V.; Tyubaeva, P. M.; Artemov, N. S.; Iordansky, A. L.

2017-12-01

In the treatment of many infectious diseases and cancer, transdermal systems based on solid polymer matrices or gels containing functional substances with antiseptic (antibacterial) properties are often used. One of the most promising types of matrices with antiseptic properties are the ones of nano- and microfiber-bonded cloth obtained by electrospinning based on biopolymer poly(3-hydroxybutyrate). The present work investigates the effects of iron (III) complex with tetraphenylporphyrin and the influence on the geometry, crystalline order and molecular dynamics in the intercrystalline (amorphous phase) of ultrathin PHB fibers.

Properties of PHA bi-, ter-, and quarter-polymers containing 4-hydroxybutyrate monomer units.

Science.gov (United States)

Zhila, Natalia; Shishatskaya, Ekaterina

2018-05-01

The present study investigates physicochemical, mechanical, and biological properties of polyhydroxyalkanoate (PHA) copolymers containing 4-hydroxybutyrate (4HB) synthesized in Cupriavidus eutrophus B10646 culture. In poly(3-hydroxybutyrate/4-hydroxybutyrate) [P(3HB/4HB)] bipolymers, 4HB varied between 10.4 and 75.0 mol%; in poly(3-hydroxybutyrate/3-hydroxyvalerate/4-hydroxybutyrate) terpolymers, 4HB constituted 28.7-55.6 mol%; and in poly(3-hydroxybutyrate/3-hydroxyvalerate/4-hydroxybutyrate/3-hydroxyhexanoate) quaterpolymers, 4HB varied between 9.3 and 13.3 mol%. The degree of crystallinity of P(3HB/4HB) copolymers decreased consistently with an increase in 4HB content, reaching 38%. The incorporation of 3-hydroxyvalerate and 3-hydroxyhexanoate into copolymers enhanced that effect. The effect of 4HB monomer units on temperature properties of copolymers was exhibited as lowering of the melting temperature and crystallization temperature, which improved the processing-related properties of the copolymers. All copolymers containing 4HB showed enhanced elongation at break compared to poly(3-hydroxybutyrate). Polymer films prepared from PHAs with different chemical composition had similar microstructure and porosity and had no toxic effect on mouse fibroblast NIH 3 T3 cells, proving their high biocompatibility. Copyright © 2018 Elsevier B.V. All rights reserved.

A fed-batch strategy to produce high poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) terpolymer yield with enhanced mechanical properties in bioreactor.

Science.gov (United States)

Aziz, Nursolehah Abd; Huong, Kai-Hee; Sipaut, Coswald Stephen; Amirul, A A

2017-11-01

This study reports an efficient fed-batch strategy to improve poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) [P(3HB-co-3HV-co-4HB)] terpolymer production by Cupriavidus sp. USMAA2-4 with enhanced mechanical properties in bioreactor. The cultivations have been performed by combining oleic acid with γ-butyrolactone at different concentration ratios with 1-pentanol at a fixed concentration. The batch and fed-batch fermentations have resulted in P(3HB-co-3HV-co-4HB) with compositions of 9-35 mol% 3HV and 4-24 mol% 4HB monomers. The DO-stat fed-batch fermentation strategies have significantly improved the production with a maximum 4.4-fold increment of cell dry weight (CDW). Besides, appropriate feeding of the substrates has resulted in an increment of terpolymer productivity from 0.086-0.347 g/L/h, with a significantly shortened cultivation time. The bacterial growth and terpolymer formation have been found to be affected by the concentration of carbon sources supplied. Characterization of P(3HB-co-3HV-co-4HB) has demonstrated that incorporation of 3HV and 4HB monomer has significantly improved the physical and thermodynamic properties of the polymers, by reducing the polymer's crystallinity. The tensile strength, Young's modulus of the terpolymer has been discovered to increase with the increase of M w . The fed-batch fermentation strategies employed in this study have resulted in terpolymers with a range of flexible materials having improved tensile strength and Young's modulus as compared to the terpolymer produced from batch fermentation. Possession of lower melting temperature indicates an enhanced thermal stability which broadens the polymer processing window.

Uniaxial drawing and mechanical properties of poly[(R)-3-hydroxybutyrate]/poly(L-lactic acid) blends.

Science.gov (United States)

Park, Jun Wuk; Doi, Yoshiharu; Iwata, Tadahisa

2004-01-01

Blends of poly(L-lactic acid) (PLLA) with two kinds of poly[(R)-3-hydroxybutyrate] (PHB) having different molecular weights, commercial-grade bacterial PHB (bacterial-PHB) and ultrahigh molecular weight PHB (UHMW-PHB), were prepared by the solvent-casting method and uniaxially drawn at two drawing temperatures, around PHB's T(g) (2 degrees C) for PHB-rich blends and around PLLA's T(g) (60 degrees C) for PLLA-rich blends. Differential scanning calorimetry analysis showed that this system was immiscible over the entire composition range. Mechanical properties of all of the samples were improved in proportion to the draw ratio. Although PLLA domains in bacterial-PHB-rich blends remained almost unstretched during cold drawing, a good interfacial adhesion between two polymers and the reinforcing role of PLLA components led to enhanced mechanical properties proportionally to the PLLA content at the same draw ratio. On the contrary, in the case of UHMW-PHB-rich blends, the minor component PLLA was found to be also oriented by cold drawing in ice water due to an increase in the interfacial entanglements caused by the very long chain length of the matrix polymer. As a result, their mechanical properties were considerably improved with increasing PLLA content compared with the bacterial-PHB system. Scanning electron microscopy observations on the surface and cross-section revealed that a layered structure with uniformly oriented microporous in the interior was obtained by selectively removal of PLLA component after simple alkaline treatment.

[From waste to treasure: turning activated sludge into bioplastic poly-3-hydroxybutyrate].

Science.gov (United States)

Chen, Jia'ni

2017-12-25

Large quantity of activated sludge is generated from wastewater treatment but without yet an appropriate deposition. High temperature can lyse the activate sludge so that nitrogen and phosphorus containing nutrients are released. Halomonas CJN was found to grow on the heat lysed activated sludge and glucose for production of bioplastic poly-3-hydroxybutyrate (PHB) in the absence of yeast extract, nitrogen and phosphorus sources as well as trace elements. This reduces the PHB production cost significantly. Furthermore, acetic acid formed from anaerobic fermentation of heat lysed activated sludge can be used to replace glucose for cell growth but not much for PHB production. After construction of an additional PHB synthesis pathway in Halomonas CJN, we can produce PHB entirely from heat lysed activated sludge, reducing production cost of PHB roughly from ¥ 30 000 Yuan/ton to ¥ 20 000 Yuan/ton, thus turning waste activated sludge to valuable raw material resource.

Two-Dimensional Raman Correlation Spectroscopy Study of Poly[(R)-3-hydroxybutyrate- co-(R)-3-hydroxyhexanoate] Copolymers.

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Noda, Isao; Roy, Anjan; Carriere, James; Sobieski, Brian J; Chase, D Bruce; Rabolt, John F

2017-07-01

Two-dimensional correlation analysis was applied to the time-dependent evolution of Raman spectra during the isothermal crystallization of bioplastic, poly[(R)-3-hydroxybutyrate- co-(R)-3-hydroxyhexanoate] or PHBHx copolymer. Simultaneous Raman measurement of both carbonyl stretching and low-frequency crystalline lattice mode regions made it possible to carry out the highly informative hetero-mode correlation analysis. The crystallization process of PHBHx involves: (1) the early nucleation stage; (2) the primary growth of well-ordered crystals of PHBHx; and (3) the secondary crystal growth phase. The latter stage probably occurs in the inter-lamellar region, with an accompanying reduction of the amorphous component, which occurs most dominantly during the primary crystal growth. The development of a fully formed lamellar structure comprising the 2 1 helices occurs after the primary growth of crystals. In the later stage, secondary inter lamellar space crystallization occurs after the full formation of packed helices comprising the lamellae.

The increase of apatite layer formation by the poly(3-hydroxybutyrate) surface modification of hydroxyapatite and β-tricalcium phosphate.

Science.gov (United States)

Szubert, M; Adamska, K; Szybowicz, M; Jesionowski, T; Buchwald, T; Voelkel, A

2014-01-01

The aim of this study was the surface modification of hydroxyapatite and β-tricalcium phosphate by poly(3-hydroxybutyrate) grafting and characterization of modificates. The bioactivity examination was carried out by the determination to grow an apatite layer on modified materials during incubation in simulated body fluid at 37°C. The additional issue taken up in this paper was to investigate the influence of fluid replacement. The process of the surface modification of biomaterials was evaluated by means of infrared and Raman spectroscopy. Formation of the apatite layer was assessed by means of scanning electron microscopy and confirmed by energy dispersive, Raman and Fourier transformed infrared spectroscopy. During exposure in simulated body fluid, the variation of the zeta potential, pH measurement and relative weight was monitored. Examination of scanning electron microscopy micrographs suggests that modification of hydroxyapatite and β-tricalcium phosphate by poly(3-hydroxybutyrate) significantly increases apatite layer formation. Raman spectroscopy evaluation revealed that the formation of the apatite layer was more significant in the case of hydroxyapatite modificate, when compared to the β-tricalcium phosphate modificate. Both modificates were characterized by stable pH, close to the natural pH of human body fluids. Furthermore, we have shown that a weekly changed, simulated body fluid solution increases apatite layer formation. © 2013.

Assessment of poly(ε-caprolactone)/poly(3-hydroxybutyrate-co-3-hydroxyvalerate) blends processed by solvent casting and electrospinning

International Nuclear Information System (INIS)

Del Gaudio, Costantino; Ercolani, Enrico; Nanni, Francesca; Bianco, Alessandra

2011-01-01

Research highlights: → PHBV, PCL and blends were processed in form of solvent cast films and e-spun mats. → A clear phase separation was observed for cast films when blended in equal amount. → E-spun blends were comprised of uniform and defect-free randomly arranged fibers. → DSC and XRD analyses demonstrated the immiscibility of PHBV and PCL. → Rearrangement of e-spun fibers and neckings, after axial test, were observed. - Abstract: Poly(ε-caprolactone) (PCL) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) were blended in different ratio, e.g. 30/70, 50/50 and 70/30 (w/w), by means of solvent casting or electrospinning. Microstructure, thermal and mechanical properties of cast films and non-woven mats were investigated by means of scanning electron microscopy (SEM), X-ray diffraction analysis (XRD), differential scanning calorimetry (DSC), and uniaxial tensile test. The microstructure of PHBV/PCL solvent cast films (thickness 65-100 μm) was strictly dependent on the composition of the blend, a clean phase separation was observed for the 50/50 (w/w) sample. All electrospun PHBV/PCL blends (thickness 350-800 μm) were characterised by uniform and homogenous fibers, the average size was about 3 μm. Both techniques led to polymeric blends comprised of separate crystalline domains associated to an amorphous interdisperse phase. It has also been demonstrated that electrospun PHBV/PCL blends showed a lower segregation degree among the crystalline domains. Solvent cast blends were characterised by superior mechanical properties in terms of tensile modulus and tensile strength compared to electrospun ones. Fractured electrospun blends showed an overall fiber rearrangement in the direction of the applied load, eventually highlighting multiple necking regions along the fibers.

Novel Poly[(R-3-Hydroxybutyrate]-Producing Bacterium Isolated from a Bolivian Hypersaline Lake

Directory of Open Access Journals (Sweden)

María Soledad Marqués-Calvo

2013-01-01

Full Text Available Poly[(R-3-hydroxybutyrate] (PHB constitutes a biopolymer synthesized from renewable resources by various microorganisms. This work focuses on finding a new PHB-producing bacterium capable of growing in conventional media used for industrial biopolymer production, its taxonomical identification, and characterization of its biopolymer. Thus, a bacterial isolation process was carried out from environmental samples of water and mud. Among the isolates, strain S29 was selected and used in a fed-batch fermentation to generate a biopolymer. This biopolymer was recovered and identified as PHB homopolymer. Surprisingly, it featured several fractions of different molecular masses, and thermal properties unusual for PHB. Hence, the microorganism S29, genetically identified as a new strain of Bacillus megaterium, proved to be interesting not only due to its growth and PHB accumulation kinetics under the investigated cultivation conditions, but also due to the thermal properties of the produced PHB.

Isolation and identification of poly beta hydroxybutyric acid accumulating bacteria of Staphylococcal sp. and characterization of biodegradable polyester.

Science.gov (United States)

Roy, Bappaditya; Banerjee, Rajat; Chatterjee, Sumana

2009-04-01

Staphylococcus sp. strain BP/SU1, capable of degrading the biopolymer and utilize it as a source of carbon and energy, was isolated from activated sludge using METABOLIX (MBX D411G). It was found that this strain was capable of accumulating poly(3-hydroxybutyric acid) P(3-HB), as granule poly (3-hydroxybutyric acid), p(3-HB), inclusion bodies when grown under suitable nutrient conditions. These strains could sustain cell growth up to a dry mass of 9.24 g/l with a doubling time of 8 to 10 hr and could accumulate P(3-HB) as granular inclusion bodies to a cell dry weight of more than 12%. P(3-HB) accumulated by this organism was isolated and characterized through NMR, FT-IR spectroscopy, UV Spectroscopy, Mass spectroscopy and Differential Scanning Calorimetry. P(3-HB) granules so isolated showed physical and chemical properties that should be possessed by a superior quality thermoplastic biopolymer.

Poly-β-hydroxybutyrate Metabolism Is Unrelated to the Sporulation and Parasporal Crystal Protein Formation in Bacillus thuringiensis.

Science.gov (United States)

Wang, Xun; Li, Zhou; Li, Xin; Qian, Hongliang; Cai, Xia; Li, Xinfeng; He, Jin

2016-01-01

Poly-3-hydroxybutyrate (PHB) is a natural polymer synthesized by many bacteria as a carbon-energy storage material. It was accumulated maximally prior to the spore formation but was degraded during the process of sporulation in Bacillus thuringiensis. Intriguingly, B. thuringiensis also accumulates large amounts of insecticidal crystal proteins (ICPs) during sporulation, which requires considerable input of carbon and energy sources. How PHB accumulation affects sporulation and ICP formation remains unclear to date. Intuitively, one would imagine that accumulated PHB provides the energy required for ICP formation. Yet our current data indicate that this is not the case. First, growth curves of the deletion mutants of phaC (encoding the PHB synthase) and phaZ (encoding the PHB depolymerase) were found to be similar to the parent strain BMB171; no difference in growth rate could be observed. In addition we further constructed the cry1Ac10 ICP gene overexpression strains of BMB171 (BMB171-cry), as well as its phaC and phaZ deletion mutants ΔphaC-cry and ΔphaZ-cry to compare their spore and ICP production rates. Again, not much change of ICP production was observed among these strains either. In fact, PHB was still degraded in most ΔphaZ-cry cells as observed by transmission electron microscopy. Together these results indicated that there is no direct association between the PHB accumulation and the sporulation and ICP formation in B. thuringiensis. Some other enzymes for PHB degradation or other energy source may be responsible for the sporulation and/or ICP formation in B. thuringiensis.

Thermoplastic starch and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) bionanocomposites before and after prolonged storage

Energy Technology Data Exchange (ETDEWEB)

Magalhaes, N.F. [Universidade Federal do Rio de Janeiro (UFRJ), Macae, RJ (Brazil); Dahmouche, K.; Andrade, C.T. [Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ (Brazil)

2012-07-01

Full text: Glycerol-plasticized cornstarch and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) were melt-mixed at a constant 70:30 (wt/wt) ratio. One-step extrusion processing were performed in the presence of a commercial organically-modified clay. The effect of increasing clay contents on the morphologies and physical properties of the blends was investigated after processing, and compared to the neat blend, by scanning electron microscopy (SEM), X-ray diffraction (XRD) and dynamic mechanical analysis (DMA). The results indicated that increasing clay contents promoted improvements in the compatibility between the components and allowed to better understand the role of clay in compatibilization mechanism. After aging for 12 months, the blends were characterized by SEM, XRD and Small-Angle X-ray Scattering (SAXS). As revealed by all techniques, the reduced size of the PHBV dispersed phase and the decreased crystallinity of both phases promoted by clay particles were maintained, although some prejudicial effect of aging was noticed. Nevertheless, SAXS results unambiguously proved the presence of both exfoliated lamellae and a small number of clay tactoids in the aged bionanocomposites. (author)

Conductive PEDOT:PSS coated polylactide (PLA) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) electrospun membranes: Fabrication and characterization

Energy Technology Data Exchange (ETDEWEB)

Chang, Hui Chung [Faculty of Bioscience and Medical Engineering, Universiti Teknologi Malaysia, 81300, Johor (Malaysia); Sun, Tao [Miniaturized Medical Devices Program, Institute of Microelectronics, Agency for Science, Technology and Research - A*STAR (Singapore); Sultana, Naznin, E-mail: naznin@biomedical.utm.my [Faculty of Bioscience and Medical Engineering, Universiti Teknologi Malaysia, 81300, Johor (Malaysia); Advanced Membrane Technology Research Center, Universiti Teknologi Malaysia, 81300, Johor (Malaysia); Lim, Mim Mim [Faculty of Bioscience and Medical Engineering, Universiti Teknologi Malaysia, 81300, Johor (Malaysia); Khan, Tareef Hayat [KALAM, Faculty of Alam Bina, Universiti Teknologi Malaysia, 81300, Johor (Malaysia); Ismail, Ahmad Fauzi [Advanced Membrane Technology Research Center, Universiti Teknologi Malaysia, 81300, Johor (Malaysia)

2016-04-01

In the current study, electrospinning technique was used to fabricate composite membranes by blending of a synthetic polymer, polylactic acid (PLA) and a natural polymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), PHBV. Conductive membranes were prepared by dipping PLA/PHBV electrospun membranes into poly(3,4-ethylenedioxythiophene)–poly(styrenesulfonate) (PEDOT:PSS) solution, which is a biocompatible polymer. The coated and uncoated membranes were evaluated using several techniques. The electrical conductivity of the coated membranes was measured using a digital multimeter. In vitro cell cytotoxicity and cell viability were measured by culturing human skin fibroblast (HSF) cells onto the membranes using MTT assays. It was observed that electrospinning of 20% (w/v) PLA/PHBV with a weight ratio of 50:50 produced the most uniform fibers with no beads. It was observed that the wettability and surface roughness of the PEDOT:PSS coated PLA/PHBV membranes were greatly increased than uncoated membrane. The results of cell viability using MTT assay, cell attachment and cell proliferation showed that the conductive PEDOT:PSS coated PLA/PHBV membrane were more favorable for tissue engineering application than their uncoated counterparts. - Highlights: • Coating with PEDOT:PSS increased the wettability of PLA/PHBV membrane. • PEDOT:PSS rendered the PLA/PHBV membrane conductive. • PEDOT:PSS coated PLA/PHBV had significantly higher cell attachment.

Conductive PEDOT:PSS coated polylactide (PLA) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) electrospun membranes: Fabrication and characterization

International Nuclear Information System (INIS)

Chang, Hui Chung; Sun, Tao; Sultana, Naznin; Lim, Mim Mim; Khan, Tareef Hayat; Ismail, Ahmad Fauzi

2016-01-01

In the current study, electrospinning technique was used to fabricate composite membranes by blending of a synthetic polymer, polylactic acid (PLA) and a natural polymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), PHBV. Conductive membranes were prepared by dipping PLA/PHBV electrospun membranes into poly(3,4-ethylenedioxythiophene)–poly(styrenesulfonate) (PEDOT:PSS) solution, which is a biocompatible polymer. The coated and uncoated membranes were evaluated using several techniques. The electrical conductivity of the coated membranes was measured using a digital multimeter. In vitro cell cytotoxicity and cell viability were measured by culturing human skin fibroblast (HSF) cells onto the membranes using MTT assays. It was observed that electrospinning of 20% (w/v) PLA/PHBV with a weight ratio of 50:50 produced the most uniform fibers with no beads. It was observed that the wettability and surface roughness of the PEDOT:PSS coated PLA/PHBV membranes were greatly increased than uncoated membrane. The results of cell viability using MTT assay, cell attachment and cell proliferation showed that the conductive PEDOT:PSS coated PLA/PHBV membrane were more favorable for tissue engineering application than their uncoated counterparts. - Highlights: • Coating with PEDOT:PSS increased the wettability of PLA/PHBV membrane. • PEDOT:PSS rendered the PLA/PHBV membrane conductive. • PEDOT:PSS coated PLA/PHBV had significantly higher cell attachment.

Thermal properties of poly(3-hydroxybutyrate)/vegetable fiber composites

Science.gov (United States)

Vitorino, Maria B. C.; Reul, Lízzia T. A.; Carvalho, Laura H.; Canedo, Eduardo L.

2015-05-01

The present work studies the thermal properties of composites of poly(3-hydroxybutyrate) (PHB) - a fully biodegradable semi-crystalline thermo-plastic obtained from renewable resources through low-impact biotechno-logical process, biocompatible and non-toxic - and vegetable fiber from the fruit (coconut) of babassu palm tree. PHB is a highly crystalline resin and this characteristic leads to suboptimal properties in some cases. Consequently, thermal properties, in particular those associated with the crystallization of the matrix, are important to judge the suitability of the compounds for specific applications. PHB/babassu composites with 0-50% load were prepared in an internal mixer. Two different types of babassu fibers with two different particle size ranges were compounded with PHB and test specimens molded by compression. Melting and crystallization behavior were studied by differential scanning calorimetry (DSC) at heating/cooling rates between 2 and 30°C/min. Several parameters, including melting point, crystallization temperature, crystallinity, and rate of crystallization, were estimated as functions of load and heating/cooling rates. Results indicate that fibers do not affect the melting process, but facilitate crystallization from the melt. Crystallization temperatures are 30 to 40°C higher for the compounds compared with the neat resin. However, the amount of fiber added has little effect on crystallinity and the degree of crystallinity is hardly affected by the load. Fiber type and initial particle size do not have a significant effect on thermal properties.

Poly(aspartic acid) (PAA) hydrolases and PAA biodegradation: current knowledge and impact on applications.

Science.gov (United States)

Hiraishi, Tomohiro

2016-02-01

Thermally synthesized poly(aspartic acid) (tPAA) is a bio-based, biocompatible, biodegradable, and water-soluble polymer that has a high proportion of β-Asp units and equivalent moles of D- and L-Asp units. Poly(aspartic acid) (PAA) hydrolase-1 and hydrolase-2 are tPAA biodegradation enzymes purified from Gram-negative bacteria. PAA hydrolase-1 selectively cleaves amide bonds between β-Asp units via an endo-type process, whereas PAA hydrolase-2 catalyzes the exo-type hydrolysis of the products of tPAA hydrolysis by PAA hydrolase-1. The novel reactivity of PAA hydrolase-1 makes it a good candidate for a biocatalyst in β-peptide synthesis. This mini-review gives an overview of PAA hydrolases with emphasis on their biochemical and functional properties, in particular, PAA hydrolase-1. Functionally related enzymes, such as poly(R-3-hydroxybutyrate) depolymerases and β-aminopeptidases, are compared to PAA hydrolases. This mini-review also provides findings that offer an insight into the catalytic mechanisms of PAA hydrolase-1 from Pedobacter sp. KP-2.

Characterization of biodegradable poly-3-hydroxybutyrate films and pellets loaded with the fungicide tebuconazole.

Science.gov (United States)

Volova, Tatiana; Zhila, Natalia; Vinogradova, Olga; Shumilova, Anna; Prudnikova, Svetlana; Shishatskaya, Ekaterina

2016-03-01

Biodegradable polymer poly(3-hydroxybutyrate) (P3HB) has been used as a matrix to construct slow-release formulations of the fungicide tebuconazole (TEB). P3HB/TEB systems constructed as films and pellets have been studied using differential scanning calorimetry, X-ray structure analysis, and Fourier transform infrared spectroscopy. TEB release from the experimental formulations has been studied in aqueous and soil laboratory systems. In the soil with known composition of microbial community, polymer was degraded, and TEB release after 35 days reached 60 and 36 % from films and pellets, respectively. That was 1.23 and 1.8 times more than the amount released to the water after 60 days in a sterile aqueous system. Incubation of P3HB/TEB films and pellets in the soil stimulated development of P3HB-degrading microorganisms of the genera Pseudomonas, Stenotrophomonas, Variovorax, and Streptomyces. Experiments with phytopathogenic fungi F. moniliforme and F. solani showed that the experimental P3HB/TEB formulations had antifungal activity comparable with that of free TEB.

Glycolysis of poly(3-hydroxybutyrate) via enzyme catalysis; Glicolise do poli(3-hidroxibutirato) por via enzimatica

Energy Technology Data Exchange (ETDEWEB)

Paula, Everton Luiz de, E-mail: everton2804@gmail.com [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Departamento de Quimica; Campos, Tiago Ferreira; Mano, Valdir [Universidade Federal de Sao Joao del-Rei (UFSJ), MG (Brazil). Departamento de Ciencias Naturais

2014-05-15

Poly(3-hydroxybutyrate), PHB, is a polymer with broad potential applications because of its biodegradability and biocompatibility. However, its high crystallinity is a limiting factor for many applications. To overcome this drawback, one strategy currently employed involves the reduction of the molecular weight of PHB with the concomitant formation of end-functionalized chains, such as those obtained via glycolysis. The glycolysis of PHB can be catalyzed by acid, base, or organometallic compounds. However, to our knowledge, there are no reports regarding PHB glycolysis catalyzed enzymatically. Among the major types of enzymes used in biocatalysis, the lipases stand out because they have the ability to catalyze reactions in both aqueous and organic media. Thus, in this study, we performed the enzymatic glycolysis of PHB using the lipase Amano PS (Pseudomonas cepacia) with ethane-1,2-diol (ethylene glycol) as the functionalizing agent. The results indicated that the glycolysis was successful and afforded hydroxyl-terminated oligomeric PHB polyols. Nuclear magnetic resonance spectra of the products showed characteristic signals for the terminal hydroxyl groups of the polyols, while thermogravimetric and differential scanning calorimetry analyses confirmed an increase in the thermal stability and a decrease in the crystallinity of the polyols compared with the starting PHB polymer, which were both attributed to the reduction in the molecular weight due to glycolysis. (author)

Production and characterization of poly-3-hydroxybutyrate from Bacillus cereus PS 10.

Science.gov (United States)

Sharma, Priyanka; Bajaj, Bijender Kumar

2015-11-01

Usage of renewable raw materials for production of fully degradable bioplastics (bacterial poly-3-hydroxybutyrate, PHB) has gained immense research impetus considering recalcitrant nature of petroleum based plastics, dwindling fossil fuel feed stocks, and associated green house gas emissions. However, high production cost of PHB is the major bottleneck for its wide range industrial applications. In current study, Bacillus cereus PS 10, a recent isolate, efficiently utilized molasses, an abundantly available by-product from sugar industries as sole carbon source for growth and PHB production. Most influential bioprocess variables i.e. molasses, pH and NH4Cl were identified based on Plackett-Burman-designed experiments. Design of experiment approach (response surface methodology) was further employed for optimization of these bioprocess variables, and an enhanced PHB yield (57.5%) was obtained. PHB produced by Bacillus cereus PS 10 was investigated using various physico-chemical approaches viz. thermogravimetric analysis, proton and carbon NMR ((1)H and (13)C) spectroscopy, melting point, elemental analysis and polarimetry for its detail characterization, and assessment for industrial application potential. Copyright © 2015 Elsevier B.V. All rights reserved.

Blends of poly(hydroxybutyrate and poly (epsilon-caprolactone obtained from melting mixture

Directory of Open Access Journals (Sweden)

Maria Cecília M. Antunes

2005-06-01

Full Text Available Poly(3-hydroxybutyrate (PHB is a thermoplastic polyester with a great potential owing to its biodegradability, bioreabsorbation and biological synthesis from a renewable source. Despite these characteristics, the applications of this polymer are very restricted due to its poor mechanical properties and thermal instability at temperatures above its melting point (around 175°C. Among the possibilities of improvement of these materials, the development of blends is a relatively fast and inexpensive option. Poly(epsilon-caprolactone (PCL is a semi-crystalline polymer that may be used as a biomaterial. It presents good mechanical properties, a low melting point (around 55 °C, and could be a good option to develop PHB blends, maintaining the biodegradability and bioreabsorption properties. The objective of the present work was to obtain blends of PHB and PCL by melting mixture in an internal mixer. The compositions varied from 0 to 30 wt% of PCL. DSC, DMA, and SEM were used to characterize the blends. The blends were found to be imiscible with no indication of interaction either the amorphous or crystalline state. The morphology shows PHB as the matrix and PCL as the dispersed phase.

Poly-3-hydroxybutyrate (PHB) production from alkylphenols, mono and poly-aromatic hydrocarbons using Bacillus sp. CYR1: A new strategy for wealth from waste.

Science.gov (United States)

Venkateswar Reddy, M; Mawatari, Yasuteru; Yajima, Yuka; Seki, Chigusa; Hoshino, Tamotsu; Chang, Young-Cheol

2015-09-01

In the present study five different types of alkylphenols, each of the two different types of mono and poly-aromatic hydrocarbons were selected for degradation, and conversion into poly-3-hydroxybutyrate (PHB) using the Bacillus sp. CYR1. Strain CYR1 showed growth with various toxic organic compounds. Degradation pattern of all the organic compounds at 100 mg/l concentration with or without addition of tween-80 were analyzed using high pressure liquid chromatography (HPLC). Strain CYR1 showed good removal of compounds in the presence of tween-80 within 3 days, but it took 6 days without addition of tween-80. Strain CYR1 showed highest PHB production with phenol (51 ± 5%), naphthalene (42 ± 4%), 4-chlorophenol (32 ± 3%) and 4-nonylphenol (29 ± 3%). The functional groups, structure, and thermal properties of the produced PHB were analyzed. These results denoted that the strain Bacillus sp. CYR1 can be used for conversion of different toxic compounds persistent in wastewaters into useable biological polyesters. Copyright © 2015 Elsevier Ltd. All rights reserved.

Melting temperature evolution of non-reorganized crystals. Poly(3-hydroxybutyrate)

International Nuclear Information System (INIS)

Righetti, Maria Cristina; Di Lorenzo, Maria Laura

2011-01-01

In the present study the correlation between the melting behaviour of poly(3-hydroxybutyrate) (PHB) original, non-reorganized crystals and the crystallinity increase during isothermal crystallization is presented and discussed. Since the reorganization processes modify the melting curve of original crystals, it is necessary to prevent and hinder all the processes that influence and increase the lamellar thickness. PHB exhibits melting/recrystallization on heating, the occurring of lamellar thickening in the solid state being excluded. The first step of the study was the identification of the scanning rate which inhibits PHB recrystallization at sufficiently high T c . For the extrapolated onset and peak temperatures of the main melting endotherm, which is connected to fusion of dominant lamellae, a double dependence on the crystallization time was found. The crystallization time at which T onset and T peak change their trends was found to correspond to the spherulite impingement time, so that the two different dependencies were put in relation with primary and secondary crystallizations respectively. The increase of both T onset and T peak at high crystallization times after spherulite impingement was considered an effect due to crystal superheating and an indication of a stabilization process of the crystalline phase. Such stabilization, which produces an increase of the melting temperature, is probably connected with the volume filling that occurs after spherulite impingement.

Miscibility and Crystallization Behavior of Poly (3- Hydroxybutyrate) and Poly (Ethylene Glycol) Blends Studied by Positron Annihilation Spectroscopy

International Nuclear Information System (INIS)

Abdel-Hady, E.E.; Hammam, A.M.

2010-01-01

Poly (3-hydroxybutyrate) (PHB) is linear stereo regular aliphatic polyester synthesized by some bacteria as a store of carbon and energy. Because of its high biocompatibility and the ability to be fully biodegraded, PHB is of special interest in medicine. To improve the physiochemical properties of PHB, Polyethylene glycol (PEG) was used for modifications of PHB. By using the chloroform as co-solvent a series of (PHB/PEG) blend with different ratio ranging from 100:0.0 (wt %) to 50:50 (wt %) was prepared by solution casting-technique. Positron Annihilation Lifetime (PAL) technique has been applied to study the effect of addition PEG on the structure of PHB. The positron annihilation lifetime measurements were performed with a conventional fast-fast coincidence system. The lifetime parameter, ι3 which represents the ortho-positronium atom (o-Ps) lifetime and I 3 which reflects the (o-Ps) intensity, give indication of the free-volume size and concentration respectively. Positron annihilation lifetime measurements showed that, ι3 increases by increasing PEG ratio until the concentration (80:20 wt %) then start to decrease by increasing PEG ratio. The obtained results are in agreement with the results of X-ray diffraction.

Formation and stability of .beta.-structure in biodegradable ultra-high-molecular-weight poly(3-hydroxybutyrate) by infrared, Raman, and quantum chemical calculation studies

Czech Academy of Sciences Publication Activity Database

Murakami, R.; Sato, H.; Dybal, Jiří; Iwata, T.; Ozaki, Y.

2007-01-01

Roč. 48, č. 9 (2007), s. 2672-2680 ISSN 0032-3861 R&D Projects: GA ČR GA203/05/0425 Grant - others:Ministry of Education, Culture, Sports, Science and Technology(JP) 18750107 Institutional research plan: CEZ:AV0Z40500505 Keywords : ultra-high-molecular-weight poly(hydroxybutyrate) * .beta.-form * Raman spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.065, year: 2007

Environmentally Friendly Compatibilizers from Soybean Oil for Ternary Blends of Poly(lactic acid-PLA, Poly(ε-caprolactone-PCL and Poly(3-hydroxybutyrate-PHB

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María Jesús Garcia-Campo

2017-11-01

Full Text Available Ternary blends of poly(lactic acid (PLA, poly(3-hydroxybutyrate (PHB and poly(ε-caprolactone (PCL with a constant weight percentage of 60%, 10% and 30% respectively were compatibilized with soybean oil derivatives epoxidized soybean oil (ESO, maleinized soybean oil (MSO and acrylated epoxidized soybean oil (AESO. The potential compatibilization effects of the soybean oil-derivatives was characterized in terms of mechanical, thermal and thermomechanical properties. The effects on morphology were studied by field emission scanning electron microscopy (FESEM. All three soybean oil-based compatibilizers led to a noticeable increase in toughness with a remarkable improvement in elongation at break. On the other hand, both the tensile modulus and strength decreased, but in a lower extent to a typical plasticization effect. Although phase separation occurred, all three soybean oil derivatives led somewhat to compatibilization through reaction between terminal hydroxyl groups in all three biopolyesters (PLA, PHB and PCL and the readily reactive groups in the soybean oil derivatives, that is, epoxy, maleic anhydride and acrylic/epoxy functionalities. In particular, the addition of 5 parts per hundred parts of the blend (phr of ESO gave the maximum elongation at break while the same amount of MSO and AESO gave the maximum toughness, measured through Charpy’s impact tests. In general, the herein-developed materials widen the potential of ternary PLA formulations by a cost effective blending method with PHB and PCL and compatibilization with vegetable oil-based additives.

Environmentally Friendly Compatibilizers from Soybean Oil for Ternary Blends of Poly(lactic acid)-PLA, Poly(ε-caprolactone)-PCL and Poly(3-hydroxybutyrate)-PHB.

Science.gov (United States)

Garcia-Campo, María Jesús; Quiles-Carrillo, Luis; Masia, Jaime; Reig-Pérez, Miguel Jorge; Montanes, Nestor; Balart, Rafael

2017-11-22

Ternary blends of poly(lactic acid) (PLA), poly(3-hydroxybutyrate) (PHB) and poly(ε-caprolactone) (PCL) with a constant weight percentage of 60%, 10% and 30% respectively were compatibilized with soybean oil derivatives epoxidized soybean oil (ESO), maleinized soybean oil (MSO) and acrylated epoxidized soybean oil (AESO). The potential compatibilization effects of the soybean oil-derivatives was characterized in terms of mechanical, thermal and thermomechanical properties. The effects on morphology were studied by field emission scanning electron microscopy (FESEM). All three soybean oil-based compatibilizers led to a noticeable increase in toughness with a remarkable improvement in elongation at break. On the other hand, both the tensile modulus and strength decreased, but in a lower extent to a typical plasticization effect. Although phase separation occurred, all three soybean oil derivatives led somewhat to compatibilization through reaction between terminal hydroxyl groups in all three biopolyesters (PLA, PHB and PCL) and the readily reactive groups in the soybean oil derivatives, that is, epoxy, maleic anhydride and acrylic/epoxy functionalities. In particular, the addition of 5 parts per hundred parts of the blend (phr) of ESO gave the maximum elongation at break while the same amount of MSO and AESO gave the maximum toughness, measured through Charpy's impact tests. In general, the herein-developed materials widen the potential of ternary PLA formulations by a cost effective blending method with PHB and PCL and compatibilization with vegetable oil-based additives.

Storage and degradation of poly-ß-hydroxybutyrate in activated sludge under aerobic conditions

DEFF Research Database (Denmark)

Dircks, Klaus; Henze, Mogens; van Loosdrecht, M.C.M.

2001-01-01

This research analyses the accumulation and degradation of poly-b-hydroxybutyrate (PHB) in experiments with pulse addition of acetate to samples of activated sludge from pilot-plant and full-scale wastewater treatment plants. The experiments are divided into two periods: a feast period defined as...

Biotechnological strategies to improve production of microbial poly-(3-hydroxybutyrate): a review of recent research work.

Science.gov (United States)

Peña, C; Castillo, T; García, A; Millán, M; Segura, D

2014-07-01

Poly-(3-hydroxybutyrate) [P(3HB)] is a polyester synthesized as a carbon and energy reserve material by a wide number of bacteria. This polymer is characterized by its thermo-plastic properties similar to plastics derived from petrochemical industry, such as polyethylene and polypropylene. Furthermore, P(3HB) is an inert, biocompatible and biodegradable material which has been proposed for several uses in medical and biomedical areas. Currently, only few bacterial species such as Cupriavidus necator, Azohydromonas lata and recombinant Escherichia coli have been successfully used for P(3HB) production at industrial level. Nevertheless, in recent years, several fermentation strategies using other microbial models such as Azotobacter vinelandii, A. chroococcum, as well as some methane-utilizing species, have been developed in order to improve the P(3HB) production and also its mean molecular weight. © 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Synthesis of Diblock copolymer poly-3-hydroxybutyrate -block-poly-3-hydroxyhexanoate [PHB-b-PHHx] by a β-oxidation weakened Pseudomonas putida KT2442

DEFF Research Database (Denmark)

Tripathi, Lakshmi; Wu, Lin-Ping; Chen, Jinchun

2012-01-01

), thermo- and mechanical analysis. NMR confirmed the existence of diblock copolymers consisting of 58 mol% PHB as the short chain length block with 42 mol% PHHx as the medium chain length block. The block copolymers had two glass transition temperatures (Tg) at 2.7°C and -16.4°C, one melting temperature...... (Tm) at 172.1°C and one cool crystallization temperature (Tc) at 69.1°C as revealed by differential scanning calorimetry (DSC), respectively. This is the first microbial short-chain-length (scl) and medium-chain-length (mcl) PHA block copolymer reported. CONCLUSIONS: It is possible to produce PHA......BACKGROUND: Block polyhydroxyalkanoates (PHA) were reported to be resistant against polymer aging that negatively affects polymer properties. Recently, more and more attempts have been directed to make PHA block copolymers. Diblock copolymers PHB-b-PHHx consisting of poly-3-hydroxybutyrate (PHB...

Intracellular delivery of poly(I:C) induces apoptosis of fibroblast-like synoviocytes via an unknown dsRNA sensor

Energy Technology Data Exchange (ETDEWEB)

Karpus, Olga N.; Hsiao, Cheng-Chih; Kort, Hanneke de; Tak, Paul P.; Hamann, Jörg, E-mail: j.hamann@amc.uva.nl

2016-08-26

Fibroblast-like synoviocytes (FLS) express functional membranous and cytoplasmic sensors for double-stranded (ds)RNA. Notably, FLS undergo apoptosis upon transfection with the synthetic dsRNA analog poly(I:C). We here studied the mechanism of intracellular poly(I:C) recognition and subsequent cell death in FLS. FLS responded similarly to poly(I:C) or 3pRNA transfection; however, only intracellular delivery of poly(I:C) induced significant cell death, accompanied by upregulation of pro-apoptotic proteins Puma and Noxa, caspase 3 cleavage, and nuclear segregation. Knockdown of the DExD/H-box helicase MDA5 did not affect the response to intracellular poly(I:C); in contrast, knockdown of RIG-I abrogated the response to 3pRNA. Knockdown of the downstream adaptor proteins IPS, STING, and TRIF or inhibition of TBK1 did not affect the response to intracellular poly(I:C), while knockdown of IFNAR blocked intracellular poly(I:C)-mediated signaling and cell death. We conclude that a so far unknown intracellular sensor recognizes linear dsRNA and induces apoptosis in FLS. - Highlights: • Intracellular poly(I:C) and 3pRNA evoke immune responses in FLS. • Only intracellular delivery of poly(I:C) induces FLS apoptosis. • FLS do not require MDA5 for their response to intracellular poly(I:C). • FLS respond to intracellular poly(I:C) independent of IPS and STING. • An unknown intracellular sensor recognizes linear dsRNA in FLS.

Polymer blends made of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) and epoxidized natural rubber: Thermal and mechanical response

Science.gov (United States)

Salim, Yoga Sugama; Han, Chan Chin; Kammer, Hans-Werner; Kumar, Sudesh; Neon, Gan Seng

2015-08-01

The ever-increasing demand of biodegradable over conventional polymers places microbial polyhydroxyalkanoates (PHA) as an ideal choice of research material for specific applications. In this study, polymer blends made of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx) and epoxidized natural rubber (ENR) were prepared using solution casting technique. The influence of ENR on thermal, morphological and mechanical properties of P(3HB-co-3HHx) was investigated. There are two glass transition (Tg) temperatures observed using differential scanning calorimeter. This indicates that P(3HB-co-3HHx) and ENR are immiscible at macroscopic level. Although the Tg of P(3HB-co-3HHx) is seen to shift toward ENR in the least manner, infrared analysis suggests that the crystal structure of P(3HB-co-3HHx) retains its conformational structure. In terms of morphology, ENR exists as droplets in P(3HB-co-3HHx)-rich phase, e.g. at ENR weight fraction (wENR) of 0.3. In dynamic mechanical analysis, all blend compositions exhibit solid-like behavior, with storage moduli larger than loss moduli, across the frequency sweep at room temperature.

The increase of apatite layer formation by the poly(3-hydroxybutyrate) surface modification of hydroxyapatite and β-tricalcium phosphate

Energy Technology Data Exchange (ETDEWEB)

Szubert, M., E-mail: mm.szubert@gmail.com [Faculty of Chemical Technology, Poznan University of Technology, Poznan (Poland); Adamska, K. [Faculty of Chemical Technology, Poznan University of Technology, Poznan (Poland); Szybowicz, M. [Faculty of Technical Physics, Poznan University of Technology, Poznan (Poland); Jesionowski, T. [Faculty of Chemical Technology, Poznan University of Technology, Poznan (Poland); Buchwald, T. [Faculty of Technical Physics, Poznan University of Technology, Poznan (Poland); Voelkel, A. [Faculty of Chemical Technology, Poznan University of Technology, Poznan (Poland)

2014-01-01

The aim of this study was the surface modification of hydroxyapatite and β-tricalcium phosphate by poly(3-hydroxybutyrate) grafting and characterization of modificates. The bioactivity examination was carried out by the determination to grow an apatite layer on modified materials during incubation in simulated body fluid at 37 °C. The additional issue taken up in this paper was to investigate the influence of fluid replacement. The process of the surface modification of biomaterials was evaluated by means of infrared and Raman spectroscopy. Formation of the apatite layer was assessed by means of scanning electron microscopy and confirmed by energy dispersive, Raman and Fourier transformed infrared spectroscopy. During exposure in simulated body fluid, the variation of the zeta potential, pH measurement and relative weight was monitored. Examination of scanning electron microscopy micrographs suggests that modification of hydroxyapatite and β-tricalcium phosphate by poly(3-hydroxybutyrate) significantly increases apatite layer formation. Raman spectroscopy evaluation revealed that the formation of the apatite layer was more significant in the case of hydroxyapatite modificate, when compared to the β-tricalcium phosphate modificate. Both modificates were characterized by stable pH, close to the natural pH of human body fluids. Furthermore, we have shown that a weekly changed, simulated body fluid solution increases apatite layer formation. - Highlights: • Surface modification of HA and β-TCP was performed by PHB grafting. • The growth of apatite layer on materials was examined in simulated body fluid (SBF). • The bioactivity of obtained materials was proved. • The replacement of SBF solution plays an important role in the process of apatite formation.

The increase of apatite layer formation by the poly(3-hydroxybutyrate) surface modification of hydroxyapatite and β-tricalcium phosphate

International Nuclear Information System (INIS)

Szubert, M.; Adamska, K.; Szybowicz, M.; Jesionowski, T.; Buchwald, T.; Voelkel, A.

2014-01-01

The aim of this study was the surface modification of hydroxyapatite and β-tricalcium phosphate by poly(3-hydroxybutyrate) grafting and characterization of modificates. The bioactivity examination was carried out by the determination to grow an apatite layer on modified materials during incubation in simulated body fluid at 37 °C. The additional issue taken up in this paper was to investigate the influence of fluid replacement. The process of the surface modification of biomaterials was evaluated by means of infrared and Raman spectroscopy. Formation of the apatite layer was assessed by means of scanning electron microscopy and confirmed by energy dispersive, Raman and Fourier transformed infrared spectroscopy. During exposure in simulated body fluid, the variation of the zeta potential, pH measurement and relative weight was monitored. Examination of scanning electron microscopy micrographs suggests that modification of hydroxyapatite and β-tricalcium phosphate by poly(3-hydroxybutyrate) significantly increases apatite layer formation. Raman spectroscopy evaluation revealed that the formation of the apatite layer was more significant in the case of hydroxyapatite modificate, when compared to the β-tricalcium phosphate modificate. Both modificates were characterized by stable pH, close to the natural pH of human body fluids. Furthermore, we have shown that a weekly changed, simulated body fluid solution increases apatite layer formation. - Highlights: • Surface modification of HA and β-TCP was performed by PHB grafting. • The growth of apatite layer on materials was examined in simulated body fluid (SBF). • The bioactivity of obtained materials was proved. • The replacement of SBF solution plays an important role in the process of apatite formation

Maintenance-energy-dependent dynamics of growth and poly(3-hydroxybutyrate) [P(3HB)] production by Azohydromonas lata MTCC 2311 using simple and renewable carbon substrates

OpenAIRE

Zafar,M.; Kumar,S.; Kumar,S.; Dhiman,A. K.; Park,H.-S.

2014-01-01

The dynamics of microbial growth and poly(3-hydroxybutyrate) [P(3HB)] production in growth/ non-growth phases of Azhohydromonas lata MTCC 2311 were studied using a maintenance-energy-dependent mathematical model. The values of calculated model kinetic parameters were: m s1 = 0.0005 h-1, k = 0.0965, µmax = 0.25 h-1 for glucose; m s1 = 0.003 h-1, k = 0.1229, µmax = 0.27 h-1 for fructose; and m s1 = 0.0076 h-1, k = 0.0694, µmax = 0.25 h-1 for sucrose. The experimental data of biomass growth, sub...

Morphology and dielectric properties of poly hydroxybutyrate(PHB)/polymethyl methacrylat(PMMA) blends with some antimicrobial applications

International Nuclear Information System (INIS)

Abou aiad, T.H.M.

2005-01-01

Poly(3-hydroxy)butyrate (PHB) is microbial polyester presenting the advantages of biodegradability and biocompatibility but suffers from several problems as it is brittle and has low abilities for some chemical modifications. To overcome such problems a reactive blending was done with a glassy acrylic polymer, poly(methyl methacrylate) (PMMA) with different compositions. The dielectric response of poly [(7)-3-hydroxybutyrate] (PHB)/(PMMA) was investigated as a function of frequency (100 Hz to 100 KHz) and at different temperatures from 20 to 160 degree C above the glass transition temperature of PHB. The results revealed the existence of a single relaxation process indicating that there is miscibility between amorphous fractions of PHB and PMMA. Subsequently a continuous decrease in relaxation strength occurred, indicating considerable changes in molecular mobility. The results provide further evidence that molecular mobility in the amorphous regions decreases significantly and this would be the reason for the embrittlement detected on ageing at room temperature. Also the morphological study of such blends don't show macro phase separation of PMMA on the surface scan which may be attributed to the compatibility in melt between PHB and PMMA particle in the PHB matrix. The biological activity of such investigated systems was tested against a representative number of pathogenic organisms by determining the minimum inhibitory concentration (MIC). The activity was found to increase by increasing the amount of PMMA in the blend

Tailoring of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) copolymers for bone tissue applications

Science.gov (United States)

Liu, Hui

Considerable scientific and technological progress has been made in formulating biomaterials based on natural and synthetic polymers for ultimate use in bone tissue scaffolding. One class of polymers that has drawn attention in recent years for bone tissue engineering application is the biodegradable polymer poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). PHBV, a microbial polymer, has gained special importance because of favorable mechanical characteristics, biological properties, highly adaptable structure, non-toxic degradation products, and minimal inflammatory response when used as scaffold. However, the lack of natural cell recognition sites on PHBV has resulted in delayed cell attachment, proliferation, differentiation, and tissue regeneration on the polymer. The primary objective of this research is to modify PHBV so as to improve the biocompatibility of the matrix. An approach was developed to prepare porous and bioactive molecule coated scaffold so as to improve the biocompatibility of PHBV matrix. We investigated the role of porosity, collagen coating, and ozone treatment of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) scaffold on cell proliferation. Based on biochemical assay, we established that maximum cell proliferation occurs on collagen coated and ozone treated porous PHBV film followed by collagen coated porous PHBV film than on porous PHBV film and the least on nonporous PHBV film. Confocal microscopy in combination with biochemical assay was used to generate 3D map of viable cell proliferation on the bulk PHBV matrix. The cells were cultivated on modified PHBV film in mineralization media containing beta-glycerol phosphate for predetermined time interval and later calcium deposits were stained with alizarin red-S assay. Atomic absorption (AA) technique was used to measure the Ca2+ content of the medium and it was found that the longer the cells were incubated in organic phosphate medium, the greater amount of Ca2+ from the medium

Cupriavidus malaysiensis sp. nov., a novel poly(3-hydroxybutyrate-co-4-hydroxybutyrate) accumulating bacterium isolated from the Malaysian environment.

Science.gov (United States)

Ramachandran, Hema; Shafie, Nur Asilla Hani; Sudesh, Kumar; Azizan, Mohamad Noor; Majid, Mohamad Isa Abdul; Amirul, Al-Ashraf Abdullah

2018-03-01

Bacterial classification on the basis of a polyphasic approach was conducted on three poly(3 hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] accumulating bacterial strains that were isolated from samples collected from Malaysian environments; Kulim Lake, Sg. Pinang river and Sg. Manik paddy field. The Gram-negative, rod-shaped, motile, non-sporulating and non-fermenting bacteria were shown to belong to the genus Cupriavidus of the Betaproteobacteria on the basis of their 16S rRNA gene sequence analyses. The sequence similarity value with their near phylogenetic neighbour, Cupriavidus pauculus LMG3413 T , was 98.5%. However, the DNA-DNA hybridization values (8-58%) and ribotyping analysis both enabled these strains to be differentiated from related Cupriavidus species with validly published names. The RiboPrint patterns of the three strains also revealed that the strains were genetically related even though they displayed a clonal diversity. The major cellular fatty acids detected in these strains included C15:0 ISO 2OH/C16:1 ω7c, hexadecanoic (16:0) and cis-11-octadecenoic (C18:1 ω7c). Their G+C contents ranged from 68.0  to 68.6 mol%, and their major isoprenoid quinone was Ubiquinone Q-8. Of these three strains, only strain USMAHM13 (= DSM 25816 = KCTC 32390) was discovered to exhibit yellow pigmentation that is characteristic of the carotenoid family. Their assembled genomes also showed that the three strains were not identical in terms of their genome sizes that were 7.82, 7.95 and 8.70 Mb for strains USMAHM13, USMAA1020 and USMAA2-4, respectively, which are slightly larger than that of Cupriavidus necator H16 (7.42 Mb). The average nucleotide identity (ANI) results indicated that the strains were genetically related and the genome pairs belong to the same species. On the basis of the results obtained in this study, the three strains are considered to represent a novel species for which the name Cupriavidus malaysiensis sp. nov. is proposed. The

Biosynthesis and characterization of diblock copolymer of p(3-hydroxypropionate)-block-p(4-hydroxybutyrate) from recombinant Escherichia coli

DEFF Research Database (Denmark)

Tripathi, Lakshmi; Wu, Linping; Meng, Dechuan

2013-01-01

Poly(4-hydroxybutyrate) (P4HB) is a highly elastic polymer, whereas poly(3-hydroxypropionate) (P3HP) is a polymer with enormous tensile strength. This study aimed to biosynthesize a block copolymer consisting of soft P4HB block with a strong P3HP block to gain unique and excellent material proper...

Processing and characterization of biocomposites Poly (3-hydroxybutyrate) / hydroxyapatite for temporary support in bone regeneration

International Nuclear Information System (INIS)

Cardoso, Cyntia Esteves; Ferreira, Willian Hermogenes; Andrade, Cristina Tristao

2011-01-01

Composite materials like biopolymer / hydroxyapatite (HA) has been attracting the interest of researchers because of its applications in medicine. These materials have been used as temporary supports during the regeneration of bone tissue. In this work, poly (3-hydroxybutyrate) (PHB) was chosen as matrix because it has good processability and biocompatibility. The HA was obtained by precipitation from calcium hydroxide and phosphoric acid, varying the speed of dripping, the synthesis temperature and time of calcination. The techniques of infrared absorption spectroscopy and X-ray diffraction (XRD) were used to characterize the samples synthesized HA. The biocomposites were prepared in an internal mixer, with compositions PHB / HA varied. After processing, the biocomposites were ground, characterized by XRD analysis and the crystallinity index. The aim of this study was to examine the variables during the synthesis of HA and its thermal treatment in order to obtain a powder of HA using a simple technique and accessible, creating good properties for the biocomposites prepared. (author)

Poly(3-hydroxybutyrate production in an integrated electromicrobial setup: Investigation under stress-inducing conditions.

Directory of Open Access Journals (Sweden)

Israa Salem Al Rowaihi

Full Text Available Poly(3-hydroxybutyrate (PHB, a biodegradable polymer, can be produced by different microorganisms. The PHB belongs to the family of polyhydroxyalkanoate (PHA that mostly accumulates as a granule in the cytoplasm of microorganisms to store carbon and energy. In this study, we established an integrated one-pot electromicrobial setup in which carbon dioxide is reduced to formate electrochemically, followed by sequential microbial conversion into PHB, using the two model strains, Methylobacterium extorquens AM1 and Cupriavidus necator H16. This setup allows to investigate the influence of different stress conditions, such as coexisting electrolysis, relatively high salinity, nutrient limitation, and starvation, on the production of PHB. The overall PHB production efficiency was analyzed in reasonably short reaction cycles typically as short as 8 h. As a result, the PHB formation was detected with C. necator H16 as a biocatalyst only when the electrolysis was operated in the same solution. The specificity of the source of PHB production is discussed, such as salinity, electricity, concurrent hydrogen production, and the possible involvement of reactive oxygen species (ROS.

Poly(3-hydroxybutyrate) production in an integrated electromicrobial setup: Investigation under stress-inducing conditions

Science.gov (United States)

Al Rowaihi, Israa Salem; Paillier, Alexis; Rasul, Shahid; Karan, Ram; Grötzinger, Stefan Wolfgang; Eppinger, Jörg

2018-01-01

Poly(3-hydroxybutyrate) (PHB), a biodegradable polymer, can be produced by different microorganisms. The PHB belongs to the family of polyhydroxyalkanoate (PHA) that mostly accumulates as a granule in the cytoplasm of microorganisms to store carbon and energy. In this study, we established an integrated one-pot electromicrobial setup in which carbon dioxide is reduced to formate electrochemically, followed by sequential microbial conversion into PHB, using the two model strains, Methylobacterium extorquens AM1 and Cupriavidus necator H16. This setup allows to investigate the influence of different stress conditions, such as coexisting electrolysis, relatively high salinity, nutrient limitation, and starvation, on the production of PHB. The overall PHB production efficiency was analyzed in reasonably short reaction cycles typically as short as 8 h. As a result, the PHB formation was detected with C. necator H16 as a biocatalyst only when the electrolysis was operated in the same solution. The specificity of the source of PHB production is discussed, such as salinity, electricity, concurrent hydrogen production, and the possible involvement of reactive oxygen species (ROS). PMID:29698424

Poly(3-hydroxybutyrate) production in an integrated electromicrobial setup: Investigation under stress-inducing conditions

KAUST Repository

Al Rowaihi, Israa Salem

2018-04-26

Poly(3-hydroxybutyrate) (PHB), a biodegradable polymer, can be produced by different microorganisms. The PHB belongs to the family of polyhydroxyalkanoate (PHA) that mostly accumulates as a granule in the cytoplasm of microorganisms to store carbon and energy. In this study, we established an integrated one-pot electromicrobial setup in which carbon dioxide is reduced to formate electrochemically, followed by sequential microbial conversion into PHB, using the two model strains, Methylobacterium extorquens AM1 and Cupriavidus necator H16. This setup allows to investigate the influence of different stress conditions, such as coexisting electrolysis, relatively high salinity, nutrient limitation, and starvation, on the production of PHB. The overall PHB production efficiency was analyzed in reasonably short reaction cycles typically as short as 8 h. As a result, the PHB formation was detected with C. necator H16 as a biocatalyst only when the electrolysis was operated in the same solution. The specificity of the source of PHB production is discussed, such as salinity, electricity, concurrent hydrogen production, and the possible involvement of reactive oxygen species (ROS).

Biosynthesis of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) by bacterial community from propylene oxide saponification wastewater residual sludge.

Science.gov (United States)

Wang, Yiwei; Zhu, Ying; Gu, Pengfei; Li, Yumei; Fan, Xiangyu; Song, Dongxue; Ji, Yan; Li, Qiang

2017-05-01

The saponification wastewater from the process of propylene oxide (PO) production is contaminated with high chemical oxygen demand (COD) and chlorine contents. Although the activated sludge process could treat the PO saponification wastewater effectively, the residual sludge was difficult to be disposed properly. In this research, microbes in PO saponification wastewater residual sludge were acclimated to produce poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) from volatile fatty acids. Through Miseq Illumina highthroughput sequencing, the bacterial community discrepancy between the original and the acclimated sludge samples were analyzed. The proportions of Bacillus, Acinetobacter, Brevundimonas and Pseudomonas, the potential PHBV-producers in the residual sludge, were all obviously increased. In the batch fermentation, the production of PHBV could achieve 4.262g/L at 300min, with the content increased from 0.04% to 23.67% of mixed liquor suspended solid (MLSS) in the acclimated sludge, and the COD of the PO saponification wastewater was also decreased in the fermentation. This work would provide an effective solution for the utilization of PO saponification wastewater residual sludge. Copyright © 2017 Elsevier B.V. All rights reserved.

Blends of ethylene-co-vinyl acetate and poly(3-hydroxybutyrate with adhesion property

Directory of Open Access Journals (Sweden)

A. de Lucas-Freile

2018-07-01

Full Text Available The structure and properties of ethylene-co-vinyl acetate (EVA and poly(3-hydroxybutyrate (PHB blends depended on their PHB content, i.e. PHB phase dominated the structure for amounts of PHB higher than 50 wt%, whereas EVA phase is dominant for PHB content lower than 50 wt%. EVA/PHB (70:30 blend showed unexpected different structure because of higher miscibility and the creation of new interfacial interactions between C=O and CH3 groups of PHB and CH3 and C=O groups of EVA, these interactions led changing of the phase structure of ethylene and vinyl acetate domains in EVA. As a consequence, improved thermal, viscoelastic and morphological properties were obtained. EVA+PHB blends containing 60 wt% or more PHB did not show tack and, interestingly, the addition of 20–30 wt% PHB enhanced the tack and displaced the maximum tack of pure EVA to lower temperature. The tack of EVA/PHB (70:30 blend was the highest among all blends because of its particular structure, fibrillation was also shown. Finally, the adhesion of EVA+PHB blends containing 20–30 wt% PHB to polypropylene (PP substrate was higher than the one of pure EVA because of the interactions between the ethylene domains in EVA phase of the blend and PP substrate surface.

Positron Lifetime as a Nano probe for Free Volume in Microbial Poly(3- Hydroxybutyrate/ Polymethylmethacrylate) Blends

International Nuclear Information System (INIS)

Abdel-Hady, E.E.; Abdel-El-Latif, R.M.; Mohamed, S.S.

2009-01-01

Poly(β-hydroxybutyrate) PHB is a bio technologically produced polyesters, highly crystalline, totally biodegradable with low versatility in mechanical properties. To overcome this problem it is performed a reactive blending producer with a glassy acrylic polymer, Poly(methylmethacrylate) (PMMA) with different concentrations. positron annihilation lifetime (PAL) spectroscopy has been used to study the effect of PMMA concentrations on the free volume hole properties of PHB. PAL spectra were analyzed using PALSF it program into three components which reflected three different morphologies of the polymer structure. The ortho positronium (ο-Ps) parameters revealed an inflection at (75/25 wt/wt) PHB/PMMA blend. Pure PHB and (75/25 wt/wt) PHB/PMMA blend have been measured as a function of temperature from -30 degree C to 90 degree C. The data clearly revealed the glass transition temperature (T g ) at zero degree C. An attempt is done to find a correlation between the electrical properties of PHB with different concentrations of PMMA and the positron annihilation parameters.

A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.

Directory of Open Access Journals (Sweden)

Richard Kelwick

Full Text Available Biopolymers, such as poly-3-hydroxybutyrate (P(3HB are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104 and RBS (native and B0034 design. P(3HB production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB.

A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.

Science.gov (United States)

Kelwick, Richard; Kopniczky, Margarita; Bower, Iain; Chi, Wenqiang; Chin, Matthew Ho Wai; Fan, Sisi; Pilcher, Jemma; Strutt, James; Webb, Alexander J; Jensen, Kirsten; Stan, Guy-Bart; Kitney, Richard; Freemont, Paul

2015-01-01

Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB).

Poly β-Hydroxybutyrate Production by Bacillus subtilis NG220 Using Sugar Industry Waste Water

Science.gov (United States)

Singh, Gulab; Kumari, Anish; Mittal, Arpana; Yadav, Anita; Aggarwal, Neeraj K.

2013-01-01

The production of poly β-hydroxybutyrate (PHB) by Bacillus subtilis NG220 was observed utilizing the sugar industry waste water supplemented with various carbon and nitrogen sources. At a growth rate of 0.14 g h−1 L−1, using sugar industry waste water was supplemented with maltose (1% w/v) and ammonium sulphate (1% w/v); the isolate produced 5.297 g/L of poly β-hydroxybutyrate accumulating 51.8% (w/w) of biomass. The chemical nature of the polymer was confirmed with nuclear magnetic resonance, Fourier transform infrared, and GC-MS spectroscopy whereas thermal properties were monitored with differential scanning calorimetry. In biodegradability study, when PHB film of the polymer (made by traditional solvent casting technique) was subjected to degradation in various natural habitats like soil, compost, and industrial sludge, it was completely degraded after 30 days in the compost having 25% (w/w) moisture. So, the present study gives insight into dual benefits of conversion of a waste material into value added product, PHB, and waste management. PMID:24027767

Biodegradation of poly(lactic acid, poly(hydroxybutyrate-co-hydroxyvalerate, poly(butylene succinate and poly(butylene adipate-co-terephthalate under anaerobic and oxygen limited thermophilic conditions

Directory of Open Access Journals (Sweden)

Jutakan Boonmee

2016-01-01

Full Text Available In order to study the biodegradation behavior of biodegradable plastics in landfill conditions, four types of biodegradable plastics including poly(lactic acid (PLA, poly(hydroxybutyrate-co-hydroxyvalerate (PHBV, poly(butylene succinate (PBS, and poly(butylene adipate-co-terephthalate (PBAT were tested by burying in sludge mixed soil medium under anaerobic and oxygen limited conditions. The experiments were operated at 52 ± 2ºC in dark conditions according to ISO15985. The degree of biodegradation after 75 days was investigated by weight loss determination, visual examination, and surface appearance by scanning electronic microscopy (SEM. Under both anaerobic and oxygen limited conditions, the complete degradation (100% weight loss was found only in PHBV after 75 days. The plastic degradations were ranked in the order of PHBV> PLA> PBS> PBAT. The percentage of weight losses were significantly different at p ≤ 0.05. However, for all studied plastics, the degradation under anaerobic and oxygen limited conditions did not significantly different at 95% confidence.

Analysis of poly-beta-hydroxybutyrate in environmental samples by GC-MS/MS.

Science.gov (United States)

Elhottová, D; Tríska, J; Petersen, S O; Santrůcková, H

2000-05-01

Application of gas chromatography-mass spectrometry (GC-MS) can significantly improve trace analyses of compounds in complex matrices from natural environments compared to gas chromatography only. A GC-MS/MS technique for determination of poly-beta-hydroxybutyrate (PHB), a bacterial storage compound, has been developed and used for analysis of two soils stored for up to 319 d, fresh samples of sewage sludge, as well as a pure culture of Bacillus megaterium. Specific derivatization of beta-hydroxybutyrate (3-OH C4:0) PHB monomer units by N-tert-butyl-dimethylsilyl-N-methyltrifluoracetamide (MTBSTFA) improved chromatographic and mass spectrometric properties of the analyte. The diagnostic fragmentation scheme of the derivates tert-butyldimethylsilyl ester and ether of beta-hydroxybutyric acid (MTBSTFA-HB) essential for the PHB identification was shown. The ion trap MS was used, therefore the scan gave the best sensitivity and with MS/MS the noise decreased, so the S/N was better and also with second fragmentation the amount of ions increased compared to SIM. The detection limit for MTBSTFA-HB by GC-MS/MS was about 10(-13) g microL(-1) of injected volume, while by GC (FID) and GC-MS (scan) it was around 10(-10) g microL(-1) of injected volume. Sensitivity of GC-MS/MS measurements of PHB in arable soil and activated sludge samples was down to 10 pg of PHB g(-1) dry matter. Comparison of MTBSTFA-HB detection in natural soil sample by GC (FID), GC-MS (scan) and by GC-MS/MS demonstrated potentials and limitations of the individual measurement techniques.

Preparation and characterization of biohybrid poly (3-hydroxybutyrate-co-3-hydroxyvalerate) based nanofibrous scaffolds

Science.gov (United States)

Kouhi, Monireh; Fathi, Mohammadhossein; Venugopal, Jayarama Reddy; Shamanian, Morteza; Ramakrishna, Seeram

2018-01-01

Development of bioengineered scaffolds for bone tissue regeneration is a growing area of research, especially those involving biodegradable electrospun nanofibers incorporated with ceramic nanoparticles, since they can mimic the extracellular matrix (ECM) of the native bone. In the current study, a biocomposite nanofibrous scaffolds consisting of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), fibrinogen (FIB) and bredigite (BR) nanoparticles was fabricated through electrospinning. The morphological, chemical and mechanical characteristics of the resultant scaffolds were studied by using field emission-scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR) and tensile tester, respectively. It was found that PHBV-FIB-BR scaffolds exhibited enhanced tensile strength and young modulus compared to PHBV and PHBV-FIB scaffolds. In addition, the measurements of the water contact angle suggested that incorporation of bredigite and fibrinogen into PHBV could improve the hydrophilicity of the composites. The results of bioactivity assessment performed in the simulated body fluid (SBF) demonstrated that the presence of the bredigite nanoparticles induced the nucleation and growth of apatite layer on the surface of PHBV-FIB-BR scaffold in SBF. Furthermore, the ion concentration changes of SBF solutions with composite scaffolds showed that PHBV-FIB-BR scaffolds released Ca and Si ions, which can stimulate osteoblast proliferation. The results of cell culture studies revealed the higher osteoblast proliferation, mineralization and differentiation on PHBV-FIB-BR and PHBV-FIB scaffolds than on PHBV. Our results suggest that PHBV-FIB-BR nanofibrous scaffold would be a promising candidate as a biocomposite nanofibrous scaffold material for tissue engineering applications.

Biocomposite coatings based on Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/calcium phosphates obtained by MAPLE for bone tissue engineering

Science.gov (United States)

Raşoga, O.; Sima, L.; Chiriţoiu, M.; Popescu-Pelin, G.; Fufǎ, O.; Grumezescu, V.; Socol, M.; Stǎnculescu, A.; Zgurǎ, I.; Socol, G.

2017-09-01

The aim of our research was to synthesize and investigate the physico-chemical and biological features of composite coatings based on poly(3-hydroxybutyrate-co-hydroxyvalerate) (PHBV) and commercial calcium phosphates (CaPs), hydroxyapatite and β-tricalcium phosphate, obtained by means of matrix assisted pulsed laser evaporation (MAPLE) technique. In this respect, laser fluence and dropcast studies were performed for pristine polymer and PHBV-CaPs composites. The microstructure of the synthesized coatings was investigated by scanning electron microscopy, while for the chemical structure and functional integrity we performed Fourier transform infrared spectroscopy comparative analysis. By using the X-ray diffraction measurements we experimentally evaluated the crystalline nature of the obtained composite materials, while relevant data regarding the hydrophilic/hydrophobic behavior of the synthesized coatings were obtained by performing static CA measurements. The biocompatibility of PHBV/CaPs coatings was evaluated by performing cellular adhesion and differentiation in vitro assays on mesenchymal stem cells.

PRODUCTION AND RECOVERY OF POLY-Β-HYDROXYBUTYRATE FROM WHEY DEGRADATION BY AZOTOBACTER

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A. Khanafari , A. Akhavan Sepahei, M. Mogharab

2006-07-01

Full Text Available Three strains of Azotobacter chroococcum were studied to produce poly-β hydroxybutyrate as a inclusion body by whey degradation. Optimum degradation whey results were obtained when using whey broth as a fermentation medium without extra salt, temperature at 35 °C and pH 7 (P<0.05. Lambda max for whey broth medium was determined probably about 400 nm. The effect of different nitrogenous rich compounds (NH4NO3, Bactopeptone, Casein, Yeast extract, Meat extract, Protease peptone and Tryptone on whey degradation showed that incorporation of nitrogenous compounds into the medium did not increase whey degradation by Azotobacter chroococcum 1723 (P<0.05. But poly-β hydroxyl-butyrate production was increased in presence Meat extract up to 75% of the cell dry weight after 48h. The addition of nitrogenous sourced (except ammonium nitrate had a positive effect on poly-β hydroxyl-butyrate production as it peaked in the presence of Meat extract and 4.43 g/L was accumulated in comparison to 0.5g at diazotrophically growing cells. Increasing the O2 values resulted by shaking at 122 rpm in decreased poly-β hydroxyl-butyrate yield form 4.43 to 0.04 g/L. The results show that this medium supports the growth of strain 1735 and also that this waste could be utilized as a carbon and nitrogen source. Production of poly-β hydroxyl-butyrate by using whey as a medium looks promising, since the use of inexpensive feed-stocks for poly-β hydroxyl-butyrate is essential if bioplastics are to become competitive products.

Radiolytic stabilization of poly(hydroxybutyrate)

International Nuclear Information System (INIS)

Santos, Renata F.S.; Araujo, Elmo S.

2007-01-01

Poly(hydroxybutyrate), PHB, is a thermoplastic polyester synthesized by many types of bacteria. PHB is of special interest in the manufacture of medical devices sterilizable by gamma radiation, because of remarkable characteristics like its great biocompatibility and biodegradability. However, ionizing radiation causes main chain scissions of PHB followed by the reduction in its molar mass. Then, the purpose of this study was to investigate the radiolytic stabilization of the structure and properties of Brazilian PHB by commercial additives used in photo and thermo-oxidative stabilization of polymers. Initially, casting solvent films were prepared adding 0.5 wt% of five different additives to the polymer system. These films were irradiated at 25 kGy and investigated viscosity-average molar mass (Mv) changes in order to select the best stabilizer. Among the tested additives only one, antioxidant type, provided a good radiostabilization. New polymeric films were prepared with the antioxidant, whose concentrations (wt%) were 0.3, 0.5, 0.7 and 1.0. In this case, the samples were submitted to doses that varied from 15 to 50 kGy. Viscometric analysis was performed in order to assess the radiation-induced main chain scissions. The G value (scissions/100eV of energy transferred to the system) was also obtained by the viscosity technique. The most effective additive, added to the polymer system at 0.5 wt%, promotes a decrease of 8.6 to 1.5 in G value at a dose range of 0 - 35 kGy. In addition, (FT-IR, NMR-1H) spectroscopic analyses on polymer system were also performed. (author)

Intracellular PHB conversion in a type II methanotroph studied by 13 C NMR

NARCIS (Netherlands)

Vecherskaya, M.; Dijkema, C.; Stams, A.J.M.

2001-01-01

Poly-g-hydroxybutyrate (PHB) formation under aerobic conditions via incorporation of [13C-2]acetate as a cosubstrate and its intracellular degradation under anaerobic conditions in a Type II methanotroph was studied by 13C NMR. During PHB synthesis in the presence of labelled acetate, low levels of

Release of the Diclofenac Sodium by Nanofibers of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate Obtained from Electrospinning and Solution Blow Spinning

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Michelle Andrade Souza

2014-01-01

Full Text Available Electrospun fibers are explored as a new system for controlled drug delivery. Novel techniques capable of obtaining polymer nanofibers have been reported in the literature. They include solution blow spinning (SBS, which is a technique to produce polymer nanofibers in the same range as electrospinning, using pressurized gas instead of high voltage. The present study investigates release characteristics of diclofenac sodium encapsulated at three concentrations (5, 10, and 20% w/v in poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV nanofibers made by electrospinning and SBS and determines the drug’s effect on fiber morphology and structural properties. PHBV nanofibers were characterized using scanning electronic microscopy, differential scanning calorimetry, and X-ray diffraction, and the release profile was examined via UV-Vis spectrophotometry. Both electrospinning and SBS encapsulated diclofenac sodium in PHBV membranes efficiently and effectively. The profile of the in vitro release of diclofenac sodium was dependent on drug concentration and temperature. The drug reduced crystallinity and increased flexibility.

Potential of Electrospun Poly(3-hydroxybutyrate/Collagen Blends for Tissue Engineering Applications

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Luca Salvatore

2018-01-01

Full Text Available In this work, tunable nonwoven mats based on poly(3-hydroxybutyrate (PHB and type I collagen (Coll were successfully produced by electrospinning. The PHB/Coll weight ratio (fixed at 100/0, 70/30, and 50/50, resp. was found to control the morphological, thermal, mechanical, and degradation properties of the mats. Increasing collagen amounts led to larger diameters of the fibers (in the approximate range 600–900 nm, while delaying their thermal decomposition (from 245°C to 262°C. Collagen also accelerated the hydrolytic degradation of the mats upon incubation in aqueous medium at 37°C for 23 days (with final weight losses of 1%, 15%, and 23% for 100/0, 70/30, and 50/50 samples, resp., as a result of increased mat wettability and reduced PHB crystallinity. Interestingly, 70/30 meshes were the ones displaying the lowest stiffness (~116 MPa; p<0.05 versus 100/0 and 50/50 meshes, while 50/50 samples had an elastic modulus comparable to that of 100/0 ones (~250 MPa, likely due to enhanced physical crosslinking of the collagen chains, at least at high protein amounts. All substrates were also found to allow for good viability and proliferation of murine fibroblasts, up to 6 days of culture. Collectively, the results evidenced the potential of as-spun PHB/Coll meshes for tissue engineering applications.

Structure of d-3-hydroxybutyrate dehydrogenase prepared in the presence of the substrate d-3-hydroxybutyrate and NAD+

International Nuclear Information System (INIS)

Hoque, Md Mominul; Shimizu, Satoru; Juan, Ella Czarina Magat; Sato, Yoshiteru; Hossain, Md Tofazzal; Yamamoto, Tamotsu; Imamura, Shigeyuki; Suzuki, Kaoru; Amano, Hitoshi; Sekiguchi, Takeshi; Tsunoda, Masaru; Takénaka, Akio

2009-01-01

The crystal structure of A. faecalisd-3-hydroxybutyrate dehydrogenase prepared in the presence of d-3-hydroxybutyrate and NAD + reveals the substrate/product-binding geometry as the first example which suggests that the catalytic reaction occurs by shuttle movements of a hydrogen negative ion from the substrate to NAD + and from NADH to the product. d-3-Hydroxybutyrate dehydrogenase from Alcaligenes faecalis catalyzes the reversible conversion between d-3-hydroxybutyrate and acetoacetate. The enzyme was crystallized in the presence of the substrate d-3-hydroxybutyrate and the cofactor NAD + at the optimum pH for the catalytic reaction. The structure, which was solved by X-ray crystallography, is isomorphous to that of the complex with the substrate analogue acetate. The product as well as the substrate molecule are accommodated well in the catalytic site. Their binding geometries suggest that the reversible reactions occur by shuttle movements of a hydrogen negative ion from the C3 atom of the substrate to the C4 atom of NAD + and from the C4 atom of NADH to the C3 atom of the product. The reaction might be further coupled to the withdrawal of a proton from the hydroxyl group of the substrate by the ionized Tyr155 residue. These structural features strongly support the previously proposed reaction mechanism of d-3-hydroxybutyrate dehydrogenase, which was based on the acetate-bound complex structure

A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry

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L.P.S. Alves

Full Text Available The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i cell permeabilization, ii Nile red staining, and iii analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99 compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry.

Science.gov (United States)

Alves, L P S; Almeida, A T; Cruz, L M; Pedrosa, F O; de Souza, E M; Chubatsu, L S; Müller-Santos, M; Valdameri, G

2017-01-16

The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.

A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry

Science.gov (United States)

Alves, L.P.S.; Almeida, A.T.; Cruz, L.M.; Pedrosa, F.O.; de Souza, E.M.; Chubatsu, L.S.; Müller-Santos, M.; Valdameri, G.

2017-01-01

The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots. PMID:28099582

Biocompatibility of plasma-treated poly(3-hydroxybutyrate-co-3-hydroxyvalerate) nanofiber mats modified by silk fibroin for bone tissue regeneration

Energy Technology Data Exchange (ETDEWEB)

Unalan, Irem [Biomedical Technologies Department, Graduate School of Natural and Applied Sciences, Dokuz Eylul University, Izmir (Turkey); Colpankan, Oylum [Metallurgical and Materials Engineering Department, Faculty of Engineering, Dokuz Eylul University, Izmir (Turkey); Albayrak, Aylin Ziylan, E-mail: aylin.albayrak@deu.edu.tr [Metallurgical and Materials Engineering Department, Faculty of Engineering, Dokuz Eylul University, Izmir (Turkey); Gorgun, Cansu [Biomedical Technologies Department, Graduate School of Natural and Applied Sciences, Ege University, Izmir (Turkey); Urkmez, Aylin Sendemir [Biomedical Technologies Department, Graduate School of Natural and Applied Sciences, Ege University, Izmir (Turkey); Bioengineering Department, Faculty of Engineering, Ege University, Izmir (Turkey)

2016-11-01

The objective of this study was to produce biocompatible plasma-treated and silk-fibroin (SF) modified poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofiber mats. The mats were plasma-treated using O{sub 2} or N{sub 2} gas to increase their hydrophilicity followed by SF immobilization for the improvement of biocompatibility. Contact angle measurements and SEM showed increased hydrophilicity and no disturbed morphology, respectively. Cell proliferation assay revealed that SF modification together with N{sub 2} plasma (PS/N{sub 2}) promoted higher osteoblastic (SaOs-2) cell viability. Although, O{sub 2} plasma triggered more mineral formation on the mats, it showed poor cell viability. Consequently, the PS/N{sub 2} nanofiber mats would be a potential candidate for bone tissue engineering applications. - Highlights: • N{sub 2}-plasma treated and silk fibroin modified mats do not show hydrophobic recovery. • Biomineralization is better on O{sub 2}-plasma treated and silk fibroin modified mats. • SaOS-2 cells like to proliferate on N{sub 2}-plasma treated surfaces.

Biosynthesis of poly(3-hydroxybutyrate) (PHB) by Cupriavidus necator H16 from jatropha oil as carbon source.

Science.gov (United States)

Batcha, Abeed Fatima Mohidin; Prasad, D M Reddy; Khan, Maksudur R; Abdullah, Hamidah

2014-05-01

Poly(3-hydroxybutyrate) (PHB) is a biodegradable polymer that can be synthesized through bacterial fermentation. In this study, Cupriavidus necator H16 is used to synthesize PHB by using Jatropha oil as its sole carbon source. Different variables mainly jatropha oil and urea concentrations, and agitation rate were investigated to determine the optimum condition for microbial fermentation in batch culture. Based on the results, the highest cell dry weight and PHB concentrations of 20.1 and 15.5 g/L, respectively, were obtained when 20 g/L of jatropha oil was used. Ethanol was used as external stress factor and the addition of 1.5 % ethanol at 38 h had a positive effect with a high PHB yield of 0.987 g PHB/g jatropha oil. The kinetic studies for cell growth rate and PHB production were conducted and the data were fitted with Logistic and Leudeking–Piret models. The rate constants were evaluated and the theoretical values were in accordance with the experimental data obtained

Poly(3-hydroxybutyrate/ZnO Bionanocomposites with Improved Mechanical, Barrier and Antibacterial Properties

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Ana M. Díez-Pascual

2014-06-01

Full Text Available Poly(3-hydroxybutyrate (PHB-based bionanocomposites incorporating different contents of ZnO nanoparticles were prepared via solution casting technique. The nanoparticles were dispersed within the biopolymer without the need for surfactants or coupling agents. The morphology, thermal, mechanical, barrier, migration and antibacterial properties of the nanocomposites were investigated. The nanoparticles acted as nucleating agents, increasing the crystallization temperature and the degree of crystallinity of the matrix, and as mass transport barriers, hindering the diffusion of volatiles generated during the decomposition process, leading to higher thermal stability. The Young’s modulus, tensile and impact strength of the biopolymer were enhanced by up to 43%, 32% and 26%, respectively, due to the strong matrix-nanofiller interfacial adhesion attained via hydrogen bonding interactions, as revealed by the FT-IR spectra. Moreover, the nanocomposites exhibited reduced water uptake and superior gas and vapour barrier properties compared to neat PHB. They also showed antibacterial activity against both Gram-positive and Gram-negative bacteria, which was progressively improved upon increasing ZnO concentration. The migration levels of PHB/ZnO composites in both non-polar and polar simulants decreased with increasing nanoparticle content, and were well below the current legislative limits for food packaging materials. These biodegradable nanocomposites show great potential as an alternative to synthetic plastic packaging materials especially for use in food and beverage containers and disposable applications.

Pseudomonas oleovorans as a source of bioplastics : production and characterization of poly(3-hydroxyalkanoates)

NARCIS (Netherlands)

Preusting, Johannes Cornelis Gerhardus

1992-01-01

Poly(3-hydroxyalkanoates) WAS) are biological polyesters which are accumulated by a wide range of bacteria under conditions of excess carbon source and limiting concentrations of an essential nutrient such as nitrogen. Poly(3-hydroxybutyrate) (PHB), the best known example of these biopolymers has

Study on formation of poly(β-hydroxybutyrate-co-hydroxyvalerate) (PHBV) fiber

Institute of Scientific and Technical Information of China (English)

Long CHEN; Peng WANG; Yanmo CHEN; Yu ZHANG; Meifang ZHU

2008-01-01

In this paper, the crystallization behavior, ther-mal degradation properties, rheological behavior and the spinnability of poly(β-hydroxybutyrate-co-hydroxyvale-rate) (PHBV) fiber were studied. Experimental results indicated that the spherulite growth rate of PHBV was very slow and its size was very large. PHBV began to degrade above 170℃. The flowing curve indicated that the processing temperature and the residential time had important effects on PHBV melts. When the equipment of melting spinning was improved and processing conditions were strictly controlled, the mechanical properties of the PHBV filament can comply with the requirements of the American Pharmacopoeia.

Composite biodegradable biopolymer coatings of silk fibroin - Poly(3-hydroxybutyric-acid-co-3-hydroxyvaleric-acid) for biomedical applications

Science.gov (United States)

Miroiu, Floralice Marimona; Stefan, Nicolaie; Visan, Anita Ioana; Nita, Cristina; Luculescu, Catalin Romeo; Rasoga, Oana; Socol, Marcela; Zgura, Irina; Cristescu, Rodica; Craciun, Doina; Socol, Gabriel

2015-11-01

Composite silk fibroin-poly(3-hydroxybutyric-acid-co-3-hydroxyvaleric-acid) (SF-PHBV) biodegradable coatings were grown by Matrix Assisted Pulsed Laser Evaporation on titanium substrates. Their physico-chemical properties and particularly the degradation behavior in simulated body fluid at 37 °C were studied as first step of applicability in local controlled release for tissue regeneration applications. SF and PHBV, natural biopolymers with excellent biocompatibility, but different biodegradability and tensile strength properties, were combined in a composite to improve their properties as coatings for biomedical uses. FTIR analyses showed the stoichiometric transfer from targets to coatings by the presence in the spectra of the main absorption maxima characteristic of both polymers. XRD investigations confirmed the FTIR results showing differences in crystallization behavior with respect to the SF and PHBV content. Contact angle values obtained through wettability measurements indicated the MAPLE deposited coatings were highly hydrophilic; surfaces turning hydrophobic with the increase of the PHBV component. Degradation assays proved that higher PHBV contents resulted in enhanced resistance and a slower degradation rate of composite coatings in SBF. Distinct drug-release schemes could be obtained by adjusting the SF:PHBV ratio to controllably tuning the coatings degradation rate, from rapid-release formulas, where SF predominates, to prolonged sustained ones, for larger PHBV content.

Physical-chemical properties of nanocomposites based on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and titanium dioxide nanoparticles

Science.gov (United States)

Braga, Natália F.; da Silva, Ana Paula; Moraes Arantes, Tatiane; Lemes, Ana Paula; Cristovan, Fernando Henrique

2018-01-01

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was reinforced with titanium dioxide (TiO2) in concentrations of 1.0%, 2.5% and 5.0% (m/m) to produce nanocomposites by the solvent casting technique. TiO2 was synthesized by a hydrothermal treatment to produce nanoparticles. The nanostructure of the nanoparticles was studied by x-ray diffraction analysis (XRD) and transmission electron microscopy (TEM). The XRD confirmed TiO2 crystalline nanoparticles, with a mixture of anatase and rutile phases. Through TEM analysis, the formation of TiO2 nanorod agglomerates with an average diameter and length of 40 and 12 nm, respectively, was observed. The thermal and mechanical properties of the pure PHBV and nanocomposite films were characterized by differential scanning calorimetry (DSC) and dynamic mechanical analysis. The DSC analysis showed that the glass transition temperature decreased with the inclusion of TiO2 in the PHBV matrix in relation to pure PHBV. The results of biodegradation assays for the PHBV and nanocomposites in an aqueous medium and in soil showed morphological and structural changes for all samples, indicating a high biodegradation rate for this material. The most important conclusion is that the biodegradation of the PHBV was not affected by the addition of nanoparticles, thus enabling the use of nanocomposites in applications requiring biodegradable materials.

Poly(3-hydroxybutyrate) and Eudragit E microparticles: a release system to enhance the aqueous solubility of felodipine and simvastatin

International Nuclear Information System (INIS)

Bazzo, Giovana C.; Pezzini, Bianca R.; Zetola, Melissa; Wagner, Theodoro M.; Caetano, Daniela B.; Boch, Maura; Schmucker, Iara C.; Schucko, Sacha K.

2009-01-01

Poor water-soluble drugs are a problem for the development of oral solid dosage forms, since it has great potential for low bioavailability. Thus, release systems that promote the increase of aqueous solubility of these drugs are advantageous. This study aimed to evaluate the feasibility of incorporation of felodipine and simvastatin in polymeric microparticles, to improve the aqueous solubility of the drugs. Microparticles of poly(3-hydroxybutyrate) [PHB] and Eudragit E was prepared by emulsion - solvent evaporation technique and characterized as to morphology and encapsulation efficiency of the drugs. Particles with spherical shapes and high levels of drug encapsulated were obtained. There was a significant increase in aqueous solubility of felodipine and simvastatin after its incorporation into the polymeric microparticles. X-ray diffraction analysis showed the conversion of both drugs to amorphous form, which may have contributed to increased the solubility. (author)

Synthesis of poly(3-hydroxybutyrate using enzymes Azospirillum brasilense in vitro

Directory of Open Access Journals (Sweden)

Soheila Abbasi

2017-06-01

Full Text Available Introduction:Polyhydroxyalkaniates (PHAS are bacterial polymers that are formed as naturally occurring storage polyesters by a wide range of microorganisms usually under unbalanced growth conditions. Mechanical properties of PHAS make them suitable replacements for petrochemically produced bulk plastics (polyethylene, polypropylene etc., but in contrast to these commodity plastics PHA are completely degradable to carbon dioxide and water through natural microbiological mineralization. PHAS can be produced by biotechnological processes under controlled conditions. Polyhydroxybotyric acid (PHB was the first of thePHAS discovered and is the most abundant polyester found in bacteria. Materials and methods: First, a rich bacterial suspension prepared from poly(3-hydroxybutyrate, then the cells were broken down and ensure the absence of live cells.The cell free enzymes were added to M9 medium.Then the sample turbidity was measured at a wavelength of 600 nm and incubated at 30°C and centrifiuged at 120rpm. Next, the amount of PHB as well as solution turbidity were measured and compared with control. The physical and chemical analyzes of extracted PHB samples such as UV absorbtion, FTIR, HPLC and HNMR assesed and compared with standard sample. Results: In twelve days this experiment was conducted, from the third day, the amount of polyhydroxybutyrate was measurable and the highest polyhydroxybutyrate was gained on the tenth day. The level of PHB was constant. Moreover, the level of glucose was decreased gradually till the tenth day. By Physico-chemical analysis, production of polyhydroxybutyrate was approved. Discussion and conclusion: In this experiment, the reaction efficiency was 37.5 percent. Probably the rest of glucose could be changed to other intermediate compounds by other enzymes in the sample.Therefore, by purification of the enzyme, usage of specific substrate and optimization of conditions outside the cell could help the production of

ZnO-reinforced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) bionanocomposites with antimicrobial function for food packaging.

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Díez-Pascual, Ana M; Díez-Vicente, Angel L

2014-06-25

Biodegradable nanocomposites were prepared by adding ZnO nanoparticles to bacterial polyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) via solution casting technique. The morphology, thermal, mechanical, antibacterial, barrier, and migration properties of the nanocomposites were analyzed. The nanoparticles were uniformly dispersed within PHBV without the aid of coupling agents, and acted effectively as nucleating agents, raising the crystallization temperature and the level of crystallinity of the matrix while decreasing its crystallite size. A gradual rise in thermal stability was found with increasing ZnO loading, since the nanofillers hinder the diffusion of volatiles generated during the decomposition process. The nanocomposites displayed superior stiffness, strength, toughness, and glass transition temperature, whereas they displayed reduced water uptake and oxygen and water vapor permeability compared to the neat biopolymer, related to the strong matrix-nanofiller interfacial adhesion attained via hydrogen bonding interactions. At an optimal concentration of 4.0 wt % ZnO, the tensile strength and Young's and storage moduli showed a maximum that coincided with the highest crystallinity and the best barrier properties. PHBV/ZnO films showed antibacterial activity against human pathogen bacteria, and the effect on Escherichia coli was stronger than on Staphylococcus aureus. The overall migration levels of the nanocomposites in both nonpolar and polar simulants dropped upon increasing nanoparticle content, and were well below the limits required by the current normative for food packaging materials. These sustainable nanomaterials with antimicrobial function are very promising to be used as containers for beverage and food products as well as for disposable applications like cutlery or overwrap films.

Controlled release profiles of dipyridamole from biodegradable microspheres on the base of poly(3-hydroxybutyrate.

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2007-12-01

Full Text Available Novel biodegradable microspheres on the base of poly(3-hydroxybutyrate (PHB designed for controlled release of antithrombotic drug, namely dipyridamole (DPD, have been kinetically studied. The profiles of release from the microspheres with different diameters 4, 9, 63, and 92 µm present the progression of nonlinear and linear stages. Diffusionkinetic equation describing both linear (PHB hydrolysis and nonlinear (diffusion stages of the DPD release profiles from the spherical subjects has been written down as the sum of two terms: desorption from the homogeneous sphere in accordance with diffusion mechanism and the zero-order release. In contrast to the diffusivity dependence on microsphere size, the constant characteristics (k of linearity are scarcely affected by the diameter of PHB microparticles. The view of the kinetic profiles as well as the low rate of DPD release are in satisfactory agreement with kinetics of weight loss measured in vitro for the PHB films. Taking into account kinetic results, we suppose that the degradation of both films and PHB microspheres is responsible for the linear stage of DPD release profiles. In the nearest future, combination of biodegradable PHB and DPD as a representative of proliferation cell inhibitors will give possibility to elaborate the novel injectable therapeutic system for a local, long-term, antiproliferative action.

Biorefinery production of poly-3-hydroxybutyrate using waste office paper hydrolysate as feedstock for microbial fermentation.

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Neelamegam, Annamalai; Al-Battashi, Huda; Al-Bahry, Saif; Nallusamy, Sivakumar

2018-01-10

Waste paper, a major fraction of municipal solid waste, has a potential to serve as renewable feedstock for the biorefineries of fuels, chemicals and materials due to rich in cellulose and abundant at low cost. This study evaluates the possibility of waste office paper (WOP) to serve as a potential feedstock for the biorefinery production of poly (3-hydroxybutyrate). In this study, the WOP was pretreated, enzymatically saccharified and the hydrolysate was used for PHB production. The hydrolysate mainly consists of glucose (22.70g/L) and xylose (1.78g/L) and the corresponding sugar yield was about 816mg/g. Ammonium sulphate and C/N ratio 20 were identified as most favorable for high yield of PHB. The batch fermentation of Cupriavidus necator using the pretreated WOP hydrolysate resulted in cell biomass, PHB production and PHB content of 7.74g/L, 4.45g/L and 57.52%, respectively. The volumetric productivity and yield achieved were 0.061g/L/h and 0.210g/g sugar, respectively. The results suggested that WOP could be a potential alternative feedstock for the biorefinery production of bioplastics. Copyright © 2017 Elsevier B.V. All rights reserved.

Characterization of poly-3-hydroxybutyrate (PHB) produced from Ralstonia eutropha using an alkali-pretreated biomass feedstock.

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Saratale, Ganesh D; Oh, Min-Kyu

2015-09-01

Alkaline pretreatment using NaOH, KOH, or NaOCl has been applied to various types of waste biomass to enhance enzymatic digestibility. Pretreatment (2% NaOH, 121 °C, 30 min) of rice paddy straw (PS) resulted in a maximum yield of 703 mg of reducing sugar per gram of PS with 84.19% hydrolysis yield after a two-step enzymatic hydrolysis process. Ralstonia eutropha ATCC 17699 was tested for its ability to synthesize poly-3-hydroxybutyrate (PHB) using PS hydrolysates as its sole carbon source. It is noteworthy that dry cell weight, polyhydroxyalkanoate (PHA) accumulation and PHB yield with the use of laboratory-grade sugars were similar to those achieved with PS-derived sugars. Under optimized conditions, we observed maximal PHA accumulation (75.45%) and PHB production (11.42 g/L) within 48 h of fermentation. After PHB recovery, the physicochemical properties of PHB were determined by various analytical techniques, showed the results were consistent with the characteristics of a standard polymer of PHB. Thus, the PS hydrolysate proved to be an excellent cheap carbon substrate for PHB production. Copyright © 2015 Elsevier B.V. All rights reserved.

Biodegradation improvement of poly(3-hydroxy-butyrate) films by entomopathogenic fungi and UV-assisted surface functionalization.

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Kessler, Felipe; Marconatto, Leticia; Rodrigues, Roberta da Silva Bussamara; Lando, Gabriela Albara; Schrank, Augusto; Vainstein, Marilene Henning; Weibel, Daniel Eduardo

2014-01-05

Ultraviolet (UV)-assisted surface modification in the presence of oxygen was used as initial step to achieve controlled degradation of poly(3-hydroxy-butyrate), PHB, films by entomopathogenic fungi. Treated surfaces were investigated by surface analysis techniques (water contact angle, Fourier Transformed Infrared Spectroscopy in Attenuated Total Reflectance mode, X-ray Photoelectron Spectroscopy, Near-edge X-ray Absorption Fine Structure, Gel Permeation Chromatography, Optical Microscopy, Scanning Electron Microscopy, and weight loss). After the UV-assisted treatments, new carbonyl groups in new chemical environments were detected by XPS and NEXAFS spectroscopy. The oxidizing atmosphere did not allow the formation of CC bonds, indicating that Norrish Type II mechanism is suppressed during or by the treatments. The higher hydrophilicity and concentration of oxygenated functional groups at the surface of the treated films possibly improved the biodegradation of the films. It was observed a clear increase in the growth of this fungus when oxygenated groups were grafted on the polymers surfaces. This simple methodology can be used to improve and control the degradation rate of PHB films in applications that require a controllable degradation rate. Copyright © 2013 Elsevier B.V. All rights reserved.

Isolated Poly(3-Hydroxybutyrate) (PHB) Granules Are Complex Bacterial Organelles Catalyzing Formation of PHB from Acetyl Coenzyme A (CoA) and Degradation of PHB to Acetyl-CoA▿

OpenAIRE

Uchino, Keiichi; Saito, Terumi; Gebauer, Birgit; Jendrossek, Dieter

2007-01-01

Poly(3-hydroxybutyrate) (PHB) granules isolated in native form (nPHB granules) from Ralstonia eutropha catalyzed formation of PHB from 14C-labeled acetyl coenzyme A (CoA) in the presence of NADPH and concomitantly released CoA, revealing that PHB biosynthetic proteins (acetoacetyl-CoA thiolase, acetoacetyl-CoA reductase, and PHB synthase) are present and active in isolated nPHB granules in vitro. nPHB granules also catalyzed thiolytic cleavage of PHB in the presence of added CoA, resulting in...

EXTRAÇÃO DE POLI(3-HIDROXIBUTIRATO, PRODUZIDO POR Cupriavidus necator, COM CARBONATO DE PROPILENO

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Luci K. M. Quines

2015-02-01

Full Text Available The environmental impact of plastic waste has attracted worldwide attention. Amid the current context of increasing concern for the environment, biodegradable plastics have been widely studied as a replacement for synthetic plastics. Poly(3-hydroxybutyrate (P(3HB is a biopolymer stored as an intracellular energy and reserve source in many microorganisms. Because it is an intracellular product, P(3HB must be extracted from the cells at the end of the culture. The purpose of this study was to investigate the effect of extraction time, heating temperature, first standing time (after filtration and extraction, second standing time (after P(3HB precipitation and solvent amount, during the process of extracting P(3HB from Cupriavidus necator DSM 545, using propylene carbonate as solvent. The extraction kinetic of P(3HB with propylene carbonate from thermally treated biomass was evaluated at different temperatures. The physical properties of the P(3HB obtained were also evaluated. In this case, P(3HB obtained at optimal conditions of recovery (98% and purity (99% was used. Results showed that temperature was the most important factor in these responses for the range of values studied (110-150 ºC.

Morphology and mechanical properties of poly(β-hydroxybutyrate)/poly(ε-caprolactone) blends controlled with cellulosic particles.

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Chen, Jianxiang; Wang, Yuankun; Yin, Zeren; Tam, Kam C; Wu, Defeng

2017-10-15

The rigid microcrystalline cellulose (MCC) particles and semi-rigid ethyl cellulose (EC) were used to control phase morphology and mechanical properties of immiscible poly(β-hydroxybutyrate) (PHB)/poly(ε-caprolactone) (PCL) blends. The interfacial properties were evaluated by the fiber retraction and contact angle methods MCC is incompatible with PHB and PCL, and dispersed independently in the two polymer phases in their blends. However, EC is more compatible with the two polymers, with a higher affinity for PCL. And EC prefers locating in PCL domains and at the phase interface. Selective localization of MCC and EC affects the mechanical properties and phase structure of PHB/PCL blends strongly. For the co-continuous samples, the presence of MCC and EC improves both the tensile and impact strengths. For the 'sea-island' ones, however, the changes of strengths depends strongly on the phase adhesion. This work will help focus efforts on moderating structure and properties of immiscible polymer blends using cellulose particles. Copyright © 2017 Elsevier Ltd. All rights reserved.

Stoichiometry and kinetics of poly-{beta}-hydroxybutyrate metabolism in aerobic, slow growing, activated sludge cultures

Energy Technology Data Exchange (ETDEWEB)

Beun, J.J.; Paletta, F.; Loosdrecht, M.C.M. Van; Heijnen, J.J.

2000-02-20

This paper discusses the poly-{beta}-hydroxybutyrate (PHB) metabolism in aerobic, slow growing, activated sludge cultures, based on experimental data and on a metabolic model. The dynamic conditions which occur in activated sludge processes were simulated in a 2-L sequencing batch reactor (SBR) by subjecting a mixed microbial population to successive periods of external substrate availability (feast period) and no external substrate availability (famine period). Under these conditions intracellular storage and consumption of PHB was observed. It appeared that in the feast period, 66% to almost 100% of the substrate consumed is used for storage of PHB, the remainder is used for growth and maintenance processes. Furthermore, it appeared that at high sludge retention time (SRT) the growth rate in the feast and famine periods was the same. With decreasing SRT the growth rate in the feast period increased relative to the growth rate in the famine period. Acetate consumption and PHB production in the feast period both proceeded with a zero-order rate in acetate and PHB concentration respectively. PHB consumption in the famine period could best be described kinetically with a nth order degradation equation in PHB concentration. The obtained results are discussed in the context of the general activated sludge models.

Draft genome sequence of Thalassobius mediterraneus CECT 5383T, a poly-beta-hydroxybutyrate producer

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Lidia Rodrigo-Torres

2016-03-01

Full Text Available Thalassobius mediterraneus is the type species of the genus Thalassobius and a member of the Roseobacter clade, an abundant representative of marine bacteria. T. mediterraneus XSM19T (=CECT 5383T was isolated from the Western Mediterranean coast near Valencia (Spain in 1989. We present here the draft genome sequence and annotation of this strain (ENA/DDBJ/NCBI accession number CYSF00000000, which is comprised of 3,431,658 bp distributed in 19 contigs and encodes 10 rRNA genes, 51 tRNA genes and 3276 protein coding genes. Relevant findings are commented, including the complete set of genes required for poly-beta-hydroxybutyrate (PHB synthesis and genes related to degradation of aromatic compounds. Keywords: Rhodobacteraceae, Roseobacter clade, PHB, Aromatic compounds

Growth and immune response of Litopenaeus vannamei fed on β-(1, 3 glucan and poly-β hydroxybutyrate

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, Sarmin

2015-05-01

Full Text Available ABSTRACT This research was aimed to examine the growth performance and non-specific immune response of Pacific white shrimp (Litopenaeus vannamei fed on the diet supplemented with β-(1,3 glucan (BG and poly-β-hydroxybutyrate (PHB as feed additives. Shrimp juvenile at an initial body weight of 2.06±0.03 g was randomly distributed into 12 units of aquaria at a density of 20 shrimps/tank and reared for 42 days. The treatments applied in this study were control (without feed additives, 1.5 g/kg BG, 10 g/kg PHB and 1,5 g/kg BG+10 g/kg PHB. Results showed that shrimp fed on 1.5 /kg BG-supplemented feed had significantly higher growth performance and non-specific immune response. Keywords: growth, shrimp, non-specific immune response, Litopenaeus vannamei  ABSTRAK Penelitian ini bertujuan untuk menguji kinerja pertumbuhan dan respons imun nonspesifik udang vaname Litopenaeus vannamei yang diberi pakan dengan penambahan feed additive berupa β-(1,3 glukan (BG dan poli-β-hidroksibutirat (PHB. Juvenil udang 2,06±0,03 g dipelihara pada 12 unit akuarium dengan empat perlakuan dan tiga ulangan, serta padat tebar 20 ekor/tank selama 42 hari pemeliharaan. Perlakuan yang diberikan dalam penelitian ini yaitu penambahan BG (1,5 g/kg, PHB (10 g/kg, dan BG (1,5 g/kg+PHB (10 g/kg, serta kontrol (tanpa penambahan feed additive. Hasil penelitian menunjukkan bahwa udang yang diberi 1,5 g/kg BG memiliki kinerja pertumbuhan dan respons imun nonspesifik yang terbaik. Kata kunci: pertumbuhan, udang, respons imun nonspesifik, Litopenaeus vannamei

Biosynthesis of poly-3-hydroxybutyrate (PHB) from glycerol by Paracoccus denitrificans in a batch bioreactor: effect of process variables.

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Kalaiyezhini, D; Ramachandran, K B

2015-01-01

In this study, the kinetics of poly-3-hydroxybutyrate (PHB) biosynthesis from glycerol by Paracoccus denitrificans DSMZ 413 were explored in a batch bioreactor. Effects of inorganic and organic nitrogen source, carbon to nitrogen ratio, and other process variables such as pH, aeration, and initial glycerol concentration on PHB production were investigated in a 2.5-L bioreactor. Yeast extract was found to be the best nitrogen source compared to several organic nitrogen sources tested. At pH 6, specific growth rate, product formation rate, and accumulation of PHB within the cell were maximum. Specific growth rate increased with increase in oxygen transfer rate, but moderate oxygen transfer rate promoted PHB production. High glycerol concentration inhibited specific product formation rate but not growth. High initial carbon/nitrogen (C/N) ratio favored PHB accumulation and its productivity. At a C/N ratio of 21.4 (mol mol(-1)), 10.7 g L(-1) of PHB corresponding to 72% of cell dry weight was attained.

Thermal degradation kinetics and isoconversional analysis of biodegradable poly(3-hydroxybutyrate)/organomodified montmorillonite nanocomposites

International Nuclear Information System (INIS)

Achilias, Dimitris S.; Panayotidou, Elpiniki; Zuburtikudis, Ioannis

2011-01-01

Poly(3-hydroxybutyrate) (PHB)/organically modified clay nanocomposites were prepared by the melt mixing method and were characterized using wide-angle X-ray diffraction. Their thermal degradation kinetics was investigated using thermogravimetric analysis at various heating rates. Further kinetic analysis was performed using isoconversional methods and the invariant kinetic parameters method was used to estimate the so-called 'true' kinetic parameters, i.e. the pre-exponential factor, A and the activation energy, E, as well as the reaction model. It was found that intercalated structures are formed and the thermal stability of the material is improved by the addition of the nano-filler. From the isoconversional analysis, it was found that the activation energy does not vary significantly with the degree of degradation denoting degradation in one step with similar values for pure PHB and for all nanocomposites. Using the invariant kinetic parameters method, it was found that the model that best describes the experimental data was that of Sestak-Berggren's with f(a) = α n (1 - α) m , where the value of n is always larger than m and is increasing with the amount of the nano-filler. The value of the 'true' activation energy was found to be about 100 kJ mol -1 for all nanocomposites and the pre-exponential factor for PHB was estimated equal to 5.35 x 10 9 min -1 . Finally, the values of the kinetic rate constant k were found to decrease with the amount of the nano-filler up to 3 wt%, while for amounts larger than 3 wt% k increased reaching a value greater than that of pure PHB for the 10 wt% nanocomposites.

Morphology and thermal properties of biodegradable poly(hydroxybutyrate-co-hydroxyvalerate)/tungsten disulphide inorganic nanotube nanocomposites

Energy Technology Data Exchange (ETDEWEB)

Silverman, Tyler [Escuela Técnica Superior de Ingenieros Industriales, Universidad Politécnica de Madrid (ETSII-UPM), José Gutiérrez Abascal 2, 28006 Madrid (Spain); Naffakh, Mohammed, E-mail: mohammed.naffakh@upm.es [Escuela Técnica Superior de Ingenieros Industriales, Universidad Politécnica de Madrid (ETSII-UPM), José Gutiérrez Abascal 2, 28006 Madrid (Spain); Marco, Carlos; Ellis, Gary [Instituto de Ciencia y Tecnología de Polímeros (ICTP-CSIC), Juan de la Cierva 3, 28006 Madrid (Spain)

2016-02-15

Promising biodegradable and renewable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) nanocomposites based on tungsten disulphide inorganic nanotubes (INT-WS{sub 2}) were efficiently prepared by a simple solution blending method. The structure, morphology, thermal stability and crystallization behavior of the nanocomposites were investigated by ultra-high field-emission scanning electron microscope (FESEM), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS) and polarized optical microscopy (POM) techniques. As previously observed in poly(3-hydroxybutyrate) (PHB) hybrid systems, the dispersion, morphology and thermal properties of PHBV/INT-WS{sub 2} nanocomposites could be tuned by the introduction of small amounts of INT-WS{sub 2}. The results revealed that a good dispersion of INT-WS{sub 2} in the PHBV matrix influenced the morphology and non-isothermal crystallization behavior of PHBV that depends on both the INT-WS{sub 2} concentration and the cooling rate. A significant enhancement in thermal stability of PHBV and a highly efficient nucleating effect of the INT-WS{sub 2} comparable to specific nucleating agents or other nano-sized fillers was observed. These observations are of importance for extending the practical applications of these biopolymer nanocomposites towards eco-friendly (e.g. sustainable packaging) and biomedical (e.g. bone tissue engineering) applications. - Highlights: • Environmentally-friendly INT-WS{sub 2} is used to produce advanced PHBV NCPs. • Novel INT-WS{sub 2} improve the thermal stability of PHBV. • INT-WS{sub 2} is effective to accelerate the crystallization of PHBV. • Ring-banded spherulites of PHBV are observed at low INT-WS{sub 2} contents. • The benefits of using INTs compared to other nanoscale fillers are highlighted.

Functionalized poly(3-hydroxybutyric acid) bodies as new in vitro biocatalysts.

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Stenger, Benjamin; Gerber, Adrian; Bernhardt, Rita; Hannemann, Frank

2018-01-01

Cytochromes P450 play a key role in the drug and steroid metabolism in the human body. This leads to a high interest in this class of proteins. Mammalian cytochromes P450 are rather delicate. Due to their localization in the mitochondrial or microsomal membrane, they tend to aggregate during expression and purification and to convert to an inactive form so that they have to be purified and stored in complex buffers. The complex buffers and low storage temperatures, however, limit the feasibility of fast, automated screening of the corresponding cytochrome P450-effector interactions, which are necessary to study substrate-protein and inhibitor-protein interactions. Here, we present the production and isolation of functionalized poly(3-hydroxybutyrate) granules (PHB bodies) from Bacillus megaterium MS941 strain. In contrast to the expression in Escherichia coli, where mammalian cytochromes P450 are associated to the cell membrane, when CYP11A1 is heterologously expressed in Bacillus megaterium, it is located on the PHB bodies. The surface of these particles provides a matrix for immobilization and stabilization of the CYP11A1 during the storage of the protein and substrate conversion. It was demonstrated that the PHB polymer basis is inert concerning the performed conversion. Immobilization of the CYP11A1 onto the PHB bodies allows freeze-drying of the complex without significant decrease of the CYP11A1 activity. This is the first lyophilization of a mammalian cytochrome P450, which allows storage over more than 18days at 4°C instead of storage at -80°C. In addition, we were able to immobilize the cytochrome P450 on the PHB bodies in vitro. In this case the expression of the protein is separated from the production of the immobilization matrix, which widens the application of this method. This article is part of a Special Issue entitled: Cytochrome P450 biodiversity and biotechnology, edited by Erika Plettner, Gianfranco Gilardi, Luet Wong, Vlada Urlacher, Jared

The Born-Oppenheimer molecular simulations of infrared spectra of crystalline poly-(R)-3-hydroxybutyrate with analysis of weak Csbnd H⋯Odbnd C hydrogen bonds

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Brela, Mateusz Z.; Boczar, Marek; Wójcik, Marek J.; Sato, Harumi; Nakajima, Takahito; Ozaki, Yukihiro

2017-06-01

In this letter we present results of study of weak Csbnd H⋯Odbnd C hydrogen bonds of crystalline poly-(R)-3-hydroxybutyrate (PHB) by using Born-Oppenheimer molecular dynamics. The polymeric structure and IR spectra of PHB result from the presence of the weak hydrogen bonds. We applied the post-molecular dynamics analysis to consider a Cdbnd O motion as indirectly involved in the hydrogen bonds. Quantization of the nuclear motion of the oxygens was done to perform detailed analysis of the strength and properties of the Cdbnd O bands involved in the weak hydrogen bonds. We have also shown the dynamic character of the weak hydrogen bond interactions.

Plasticized poly(lactic acid)-poly(hydroxybutyrate) (PLA-PHB) blends incorporated with catechin intended for active food-packaging applications.

Science.gov (United States)

Arrieta, Marina Patricia; Castro-López, María del Mar; Rayón, Emilio; Barral-Losada, Luis Fernando; López-Vilariño, José Manuel; López, Juan; González-Rodríguez, María Victoria

2014-10-15

Active biobased packaging materials based on poly(lactic acid)-poly(hydroxybutyrate) (PLA-PHB) blends were prepared by melt blending and fully characterized. Catechin incorporation, as antioxidant compound, enhanced the thermal stability, whereas its release was improved by the addition of acetyl(tributyl citrate) (ATBC) as plasticizer. Whereas the incorporation of ATBC resulted in a reduction of elastic modulus and hardness, catechin addition produced more rigid materials due to hydrogen-bonding interactions between catechin hydroxyl groups and carbonyl groups of PLA and PHB. The quantification of catechin released into a fatty food simulant and the antioxidant effectiveness after the release process were demonstrated. The effect of the materials' exposure to a food simulant was also investigated. PHB-added materials maintained their structural and mechanical properties after 10 days in a test medium that represents the worst foreseeable conditions of the intended use. Thus, plasticized PLA-PHB blends with catechin show their potential as biobased active packaging for fatty food.

Production of green biodegradable plastics of poly(3-hydroxybutyrate) from renewable resources of agricultural residues.

Science.gov (United States)

Dahman, Yaser; Ugwu, Charles U

2014-08-01

This work describes potential opportunities for utilization of agro-industrial residues to produce green biodegradable plastics of poly(3-hydroxybutyrate) (PHB). Wheat straws were examined with good efficacy of carbon substrates using Cupriavidus necator. Production was examined in separate hydrolysis and fermentation (SHF) in the presence and absence of WS hydrolysis enzymes, and in simultaneous saccharification and fermentation (SSF) with enzymes. Results showed that production of PHB in SSF was more efficient in terms of viable cell count, cell dry weight, and PHB production and yield compared to those of SHF and glucose-control cultures. While glucose control experiment produced 4.6 g/L PHB; SSF produced 10.0 g/L compared to 7.1 g/L in SHF when utilizing enzymes during WS hydrolysis. Results showed that most of sugars produced during the hydrolysis were consumed in SHF (~98 %) compared to 89.2 % in SSF. Results also demonstrated that a combination of glucose and xylose can compensate for the excess carbon required for enhancing PHB production by C. necator. However, higher concentration of sugars at the beginning of fermentation in SHF can lead to cell inhibition and consequently catabolite repressions. Accordingly, results demonstrated that the gradual release of sugars in SSF enhanced PHB production. Moreover, the presence of sugars other than glucose and xylose can eliminate PHB degradation in medium of low carbon substrate concentrations in SSF.

Development of a sirolimus-eluting poly (L-lactide)/poly(4-hydroxybutyrate) absorbable stent for peripheral vascular intervention.

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Grabow, Niels; Bünger, Carsten M; Kischkel, Sabine; Timmermann, J Hinrich; Reske, Thomas; Martin, David P; Williams, Simon F; Schareck, Wolfgang; Sternberg, Katrin; Schmitz, Klaus-Peter

2013-10-01

Fully absorbable drug-eluting stent platforms are currently entering the clinical arena for the interventional treatment of coronary artery disease. This new technology also holds potential for application in peripheral vascular settings. Our study reports on the development of a sirolimus- (SIR) eluting absorbable polymer stent made from a blend of poly(l-lactide) and poly(4-hydroxybutyrate) (PLLA/P4HB) for peripheral vascular intervention. Stent prototypes were laser-cut from PLLA/P4HB tubes (I.D.=2.2 mm, t=250 µm), spray-coated with different PLLA/P4HB/SIR solutions, and bench-tested to determine expansion properties, fatigue, trackability and in vitro drug release kinetics. The stent prototypes were expanded with a 5.0 × 20 mm balloon catheter, and exhibited a recoil of 3.6% upon balloon deflation. Stent collapse pressure of 0.4 bar (300 mm Hg) was measured under external pressure load. Sustained scaffolding properties were observed in vitro over 14 weeks of radial fatigue loading (50 ± 25 mm Hg at 1.2 Hz). Trackability was demonstrated in bench tests with an 8 French contralateral introducer sheath. SIR release kinetics were adjusted over a broad range by varying the PLLA/P4HB ratio of the coating matrix. The newly developed absorbable SIR-eluting PLLA/P4HB stent successfully fulfilled the requirements for peripheral vascular intervention under in vitro conditions.

Maintenance-energy-dependent dynamics of growth and poly(3-hydroxybutyrate [P(3HB] production by Azohydromonas lata MTCC 2311 using simple and renewable carbon substrates

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M. Zafar

2014-06-01

Full Text Available The dynamics of microbial growth and poly(3-hydroxybutyrate [P(3HB] production in growth/ non-growth phases of Azhohydromonas lata MTCC 2311 were studied using a maintenance-energy-dependent mathematical model. The values of calculated model kinetic parameters were: m s1 = 0.0005 h-1, k = 0.0965, µmax = 0.25 h-1 for glucose; m s1 = 0.003 h-1, k = 0.1229, µmax = 0.27 h-1 for fructose; and m s1 = 0.0076 h-1, k = 0.0694, µmax = 0.25 h-1 for sucrose. The experimental data of biomass growth, substrate consumption, and P(3HB production on different carbon substrates were mathematically fitted using non-linear least square optimization technique and similar trends, but different levels were observed at varying initial carbon substrate concentration. Further, on the basis of substrate assimilation potential, cane molasses was used as an inexpensive and renewable carbon source for P(3HB production. Besides, the physico-chemical, thermal, and material properties of synthesized P(3HB were determined which reveal its suitability in various applications.

ACCUMULATION OF POLY-B-HYDROXYBUTYRATE IN A METHANE- ENRICHED, HALOGENATED, HYDROCARBON-DEGRADING SOIL COLUMN: IMPLICATIONS FOR MICROBIAL COMMUNITY STRUCTURE AND NUTRITIONAL STATUS

Science.gov (United States)

The prokarotic, endogenous storage polymer poly--hydroxybutyrate (PHB) accumulated in soil from a methane-enriched, halogenated hydrocarbon-degrading soil column. Based on phospholipid ester-linked fatty acid (PLFA) profiles, this mocrocosm has been previously reported to be sign...

Degradation of microbial polyesters.

Science.gov (United States)

Tokiwa, Yutaka; Calabia, Buenaventurada P

2004-08-01

Microbial polyhydroxyalkanoates (PHAs), one of the largest groups of thermoplastic polyesters are receiving much attention as biodegradable substitutes for non-degradable plastics. Poly(D-3-hydroxybutyrate) (PHB) is the most ubiquitous and most intensively studied PHA. Microorganisms degrading these polyesters are widely distributed in various environments. Although various PHB-degrading microorganisms and PHB depolymerases have been studied and characterized, there are still many groups of microorganisms and enzymes with varying properties awaiting various applications. Distributions of PHB-degrading microorganisms, factors affecting the biodegradability of PHB, and microbial and enzymatic degradation of PHB are discussed in this review. We also propose an application of a new isolated, thermophilic PHB-degrading microorganism, Streptomyces strain MG, for producing pure monomers of PHA and useful chemicals, including D-3-hydroxycarboxylic acids such as D-3-hydroxybutyric acid, by enzymatic degradation of PHB.

Siliceous mesostructured cellular foams/ poly(3-hydroxybutyrate-co-3-hydroxyhexanoate composite biomaterials for bone regeneration

Directory of Open Access Journals (Sweden)

Yang S

2014-10-01

Full Text Available Shengbing Yang,1,* Shuogui Xu,2,* Panyu Zhou,2,* Jing Wang,3 Honglue Tan,4 Yang Liu,5 TingTing Tang,4 ChangSheng Liu1,3,5 1The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People’s Republic of China; 2Changhai Hospital, Department of Orthopedics, the Second Military Medical University, Shanghai, People’s Republic of China; 3Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai, People’s Republic of China; 4Shanghai Key Laboratory of Orthopedic Implant, Department of Orthopedic Surgery, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine China, Shanghai, People’s Republic of China; 5Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai, People’s Republic of China *These authors contributed equally to this workAbstract: Osteoinductive and biodegradable composite biomaterials for bone regeneration were prepared by combining poly(3-hydroxybutyrate-co-3-hydroxyhexanoate (PHBHHx with siliceous mesostructured cellular foams (SMC, using the porogen leaching method. Surface hydrophilicity, morphology, and recombinant human bone morphogenetic protein 2 adsorption/release behavior of the SMC/PHBHHx scaffolds were analyzed. Results of scanning electron microscopy indicated that the SMC was uniformly dispersed in the PHBHHx scaffolds, and SMC modification scaffolds have an interconnected porous architecture with pore sizes ranging from 200 to 400 µm. The measurements of the water contact angles suggested that the incorporation of SMC into PHBHHx improves the hydrophilicity of the composite. In vitro studies with simulated body fluid show great improvements to bioactivity and biodegradability versus pure PHBHHx scaffolds. Cell adhesion and cell proliferation on the scaffolds was also evaluated, and the new

Ectopic bone formation in nude rats using human osteoblasts seeded poly(3)hydroxybutyrate embroidery and hydroxyapatite-collagen tapes constructs.

Science.gov (United States)

Mai, Ronald; Hagedorn, Manolo Gunnar; Gelinsky, Michael; Werner, Carsten; Turhani, Dritan; Späth, Heike; Gedrange, Tomas; Lauer, Günter

2006-09-01

The aim of this study was to evaluate the ectopic bone formation using tissue engineered cell-seeded constructs with two different scaffolds and primary human maxillary osteoblasts in nude rats over an implantation period of up to 96 days. Collagen I-coated Poly(3)hydroxybutyrate (PHB) embroidery and hydroxyapatite (HAP) collagen tapes were seeded with primary human maxillary osteoblasts (hOB) and implanted into athymic rnu/run rats. A total of 72 implants were placed into the back muscles of 18 rats. 24, 48 and 96 days after implantation, histological and histomorphometric analyses were made. The osteoblastic character of the cells was confirmed by immunocytochemistry and RT-PCR for osteocalcin. Histological analysis demonstrated that all cell-seeded constructs induced ectopic bone formation after 24, 48 and 96 days of implantation. There was more mineralized tissue in PHB constructs than in HAP-collagen tapes (at day 24; p embroidery or HAP-collagen tapes can induce ectopic bone formation. However, the amount of bone formed decreased with increasing length of implantation.

Vascular Endothelial Growth Factor Improves Physico-Mechanical Properties and Enhances Endothelialization of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/Poly(ε-caprolactone) Small-Diameter Vascular Grafts In vivo.

Science.gov (United States)

Antonova, Larisa V; Sevostyanova, Victoria V; Kutikhin, Anton G; Mironov, Andrey V; Krivkina, Evgeniya O; Shabaev, Amin R; Matveeva, Vera G; Velikanova, Elena A; Sergeeva, Evgeniya A; Burago, Andrey Y; Vasyukov, Georgiy Y; Glushkova, Tatiana V; Kudryavtseva, Yuliya A; Barbarash, Olga L; Barbarash, Leonid S

2016-01-01

The combination of a natural polymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and a synthetic hydrophobic polymer poly(ε-caprolactone) (PCL) is promising for the preparation of biodegradable and biocompatible small-diameter vascular grafts for bypass surgery. However, physico-mechanical properties and endothelialization rate of PHBV/PCL grafts are poor. We suggested that incorporation of vascular endothelial growth factor (VEGF) into PHBV/PCL grafts may improve their physico-mechanical properties and enhance endothelialization. Here we compared morphology, physico-mechanical properties, and in vivo performance of electrospun small-diameter vascular grafts prepared from PHBV/PCL with and without VEGF. Structure of the graft surface and physico-mechanical properties were examined by scanning electron microscopy and universal testing machine, respectively. Grafts were implanted into rat abdominal aorta for 1, 3, and 6 months with the further histological, immunohistochemical, and immunofluorescence examination. PHBV/PCL grafts with and without VEGF were highly porous and consisted mostly of nanoscale and microscale fibers, respectively. Mean pore diameter and mean pore area were significantly lower in PHBV/PCL/VEGF compared to PHBV/PCL grafts (1.47 μm and 10.05 μm(2); 2.63 μm and 47.13 μm(2), respectively). Durability, elasticity, and stiffness of PHBV/PCL grafts with VEGF were more similar to internal mammary artery compared to those without, particularly 6 months postimplantation. Both qualitative examination and quantitative image analysis showed that three-fourths of PHBV/PCL grafts with VEGF were patent and had many CD31-, CD34-, and vWF-positive cells at their inner surface. However, all PHBV/PCL grafts without VEGF were occluded and had no or a few CD31-positive cells at the inner surface. Therefore, VEGF enhanced endothelialization and improved graft patency at all the time points in a rat abdominal aorta replacement model. In conclusion, PHBV

MonoMax Suture: A New Long-Term Absorbable Monofilament Suture Made from Poly-4-Hydroxybutyrate

Directory of Open Access Journals (Sweden)

Erich K. Odermatt

2012-01-01

Full Text Available A long-term absorbable monofilament suture was developed using poly-4-hydroxybutyrate (P4HB made from a biosynthetically produced homopolymer of the natural metabolite 4-hydroxybutyrate. The suture, called MonoMax, has prolonged strength retention. At 12 weeks, a size 3-0 MonoMax suture retains approximately 50% of its initial tensile strength in vivo and is substantially degraded in one year with minimal tissue reaction. In contrast, PDS II monofilament suture (Ethicon, Inc., Somerville, NJ has no residual strength in vivo after 12 weeks. In vivo, the MonoMax suture is hydrolyzed primarily by bulk hydrolysis, and is then degraded via the Krebs cycle. MonoMax is substantially more compliant than other monofilament sutures, and incorporates an element of elasticity. Its tensile modulus of 0.48 GPa is approximately one-third of the value of the PDS II fiber providing an exceptionally flexible and pliable fiber with excellent knot strength and security. These features are further enhanced by the fiber's elasticity, which also improves knot security and may help prevent wound dehiscence. Because of its performance advantages, this suture may find clinical utility in applications where prolonged strength retention, and greater flexibility are required, particularly in procedures like abdominal wall closure where wound dehiscence is still a significant post-surgical complication.

Production of high levels of poly-3-hydroxybutyrate in plastids of Camelina sativa seeds.

Science.gov (United States)

Malik, Meghna R; Yang, Wenyu; Patterson, Nii; Tang, Jihong; Wellinghoff, Rachel L; Preuss, Mary L; Burkitt, Claire; Sharma, Nirmala; Ji, Yuanyuan; Jez, Joseph M; Peoples, Oliver P; Jaworski, Jan G; Cahoon, Edgar B; Snell, Kristi D

2015-06-01

Poly-3-hydroxybutyrate (PHB) production in plastids of Camelina sativa seeds was investigated by comparing levels of polymer produced upon transformation of plants with five different binary vectors containing combinations of five seed-specific promoters for expression of transgenes. Genes encoding PHB biosynthetic enzymes were modified at the N-terminus to encode a plastid targeting signal. PHB levels of up to 15% of the mature seed weight were measured in single sacrificed T1 seeds with a genetic construct containing the oleosin and glycinin promoters. A more detailed analysis of the PHB production potential of two of the best performing binary vectors in a Camelina line bred for larger seed size yielded lines containing up to 15% polymer in mature T2 seeds. Transmission electron microscopy showed the presence of distinct granules of PHB in the seeds. PHB production had varying effects on germination, emergence and survival of seedlings. Once true leaves formed, plants grew normally and were able to set seeds. PHB synthesis lowered the total oil but not the protein content of engineered seeds. A change in the oil fatty acid profile was also observed. High molecular weight polymer was produced with weight-averaged molecular weights varying between 600 000 and 1 500 000, depending on the line. Select lines were advanced to later generations yielding a line with 13.7% PHB in T4 seeds. The levels of polymer produced in this study are the highest reported to date in a seed and are an important step forward for commercializing an oilseed-based platform for PHB production. © 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

Development and characterization of coaxially electrospun gelatin coated poly (3-hydroxybutyric acid) thin films as potential scaffolds for skin regeneration

International Nuclear Information System (INIS)

Nagiah, Naveen; Madhavi, Lakshmi; Anitha, R.; Anandan, C.; Srinivasan, Natarajan Tirupattur; Sivagnanam, Uma Tirichurapalli

2013-01-01

The morphology of fibers synthesized through electrospinning has been found to mimic extracellular matrix. Coaxially electrospun fibers of gelatin (sheath) coated poly (3-hydroxybutyric acid) (PHB) (core) was developed using 2,2,2 trifluoroethanol(TFE) and 1,1,1,3,3,3 hexafluoro-2-propanol(HFIP) as solvents respectively. The coaxial structure and coating of gelatin with PHB fibers was confirmed through transmission electron microscopy (TEM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Thermal stability of the coaxially electrospun fibers was analyzed using thermogravimetric analysis(TGA), differential scanning calorimetry(DSC) and differential thermogravimetric analysis(DTA). Complete evaporation of solvent and gelatin grafting over PHB fibers was confirmed through attenuated total reflection-Fourier transformed infrared spectroscopy (ATR-FTIR). The coaxially electrospun fibers exhibited competent tensile properties for skin regeneration with high surface area and porosity. In vitro degradation studies proved the stability of fibers and its potential applications in tissue engineering. The fibers supported the growth of human dermal fibroblasts and keratinocytes with normal morphology indicating its potential as a scaffold for skin regeneration. - Highlights: • Coaxial electrospinning was employed to develop core-shell fibers of PHB and gelatin. • The scaffold has competent physicochemical properties. • Developed scaffold will have high impact as a dermal substitute in skin regeneration

Development and characterization of coaxially electrospun gelatin coated poly (3-hydroxybutyric acid) thin films as potential scaffolds for skin regeneration

Energy Technology Data Exchange (ETDEWEB)

Nagiah, Naveen [Bioproducts Lab, Central Leather Research Institute, Adyar, Chennai (India); Madhavi, Lakshmi; Anitha, R. [CavinKare Research Centre, Ekkattuthangal, Chennai (India); Anandan, C. [National Aerospace Laboratory, Banglore (India); Srinivasan, Natarajan Tirupattur, E-mail: naveen.nagiah@gmail.com [Conducting Polymers Lab, Department of Physics, Indian Institute of Technology Madras, Chennai (India); Sivagnanam, Uma Tirichurapalli, E-mail: suma67@gmail.com [Bioproducts Lab, Central Leather Research Institute, Adyar, Chennai (India)

2013-10-01

The morphology of fibers synthesized through electrospinning has been found to mimic extracellular matrix. Coaxially electrospun fibers of gelatin (sheath) coated poly (3-hydroxybutyric acid) (PHB) (core) was developed using 2,2,2 trifluoroethanol(TFE) and 1,1,1,3,3,3 hexafluoro-2-propanol(HFIP) as solvents respectively. The coaxial structure and coating of gelatin with PHB fibers was confirmed through transmission electron microscopy (TEM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Thermal stability of the coaxially electrospun fibers was analyzed using thermogravimetric analysis(TGA), differential scanning calorimetry(DSC) and differential thermogravimetric analysis(DTA). Complete evaporation of solvent and gelatin grafting over PHB fibers was confirmed through attenuated total reflection-Fourier transformed infrared spectroscopy (ATR-FTIR). The coaxially electrospun fibers exhibited competent tensile properties for skin regeneration with high surface area and porosity. In vitro degradation studies proved the stability of fibers and its potential applications in tissue engineering. The fibers supported the growth of human dermal fibroblasts and keratinocytes with normal morphology indicating its potential as a scaffold for skin regeneration. - Highlights: • Coaxial electrospinning was employed to develop core-shell fibers of PHB and gelatin. • The scaffold has competent physicochemical properties. • Developed scaffold will have high impact as a dermal substitute in skin regeneration.

Selective Advantage of a Spirillum sp. in a Carbon-limited Environment. Accumulation of Poly-β-hydroxybutyric Acid and Its Role in Starvation

NARCIS (Netherlands)

Matin, A.; Veldhuis, C.; Stegeman, V.; Veenhuis, M.

1979-01-01

A freshwater Spirillum sp., which apparently belongs to a niche of low nutritional status, accumulated poly-β-hydroxybutyric acid (PHB) during lactate-limited growth in continuous culture. The PHB content varied in a complex manner with the dilution rate (D), but was greatest at the lowest D value

Molecular Characterization of the Bacterial Community in Biofilms for Degradation of Poly(3-Hydroxybutyrate-co-3-Hydroxyhexanoate) Films in Seawater.

Science.gov (United States)

Morohoshi, Tomohiro; Ogata, Kento; Okura, Tetsuo; Sato, Shunsuke

2018-03-29

Microplastics are fragmented pieces of plastic in marine environments, and have become a serious environmental issue. However, the dynamics of the biodegradation of plastic in marine environments have not yet been elucidated in detail. Polyhydroxyalkanoates (PHAs) are biodegradable polymers that are synthesized by a wide range of microorganisms. One of the PHA derivatives, poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) has flexible material properties and a low melting temperature. After an incubation in seawater samples, a significant amount of biofilms were observed on the surfaces of PHBH films, and some PHBH films were mostly or partially degraded. In the biofilms that formed on the surfaces of unbroken PHBH films, the most dominant operational taxonomic units (OTUs) showed high similarity with the genus Glaciecola in the family Alteromonadaceae. On the other hand, the dominant OTUs in the biofilms that formed on the surfaces of broken PHBH films were assigned to the families Rhodobacteraceae, Rhodospirillaceae, and Oceanospirillaceae, and the genus Glaciecola mostly disappeared. The bacterial community in the biofilms on PHBH films was assumed to have dynamically changed according to the progression of degradation. Approximately 50 colonies were isolated from the biofilm samples that formed on the PHBH films and their PHBH-degrading activities were assessed. Two out of three PHBH-degrading isolates showed high similarities to Glaciecola lipolytica and Aestuariibacter halophilus in the family Alteromonadaceae. These results suggest that bacterial strains belonging to the family Alteromonadaceae function as the principal PHBH-degrading bacteria in these biofilms.

Structural and morphological studies on poly(3-hydroxybutyrate acid) (PHB)/chitosan drug releasing microspheres prepared by both single and double emulsion processes

Energy Technology Data Exchange (ETDEWEB)

Shih, W.-J. [Department of Materials Science and Engineering, National Cheng Kung University, 1 Ta-Hsueh Road, Tainan 70101, Taiwan (China); Chen, Y.-H. [Department of Mechanical Engineering, National Kaohsiung University of Applied Sciences, 415 Chien-kung Road, Kaohsiung 80782, Taiwan (China); Shih, C.-J. [Faculty of Fragrance and Cosmetics, Kaohsiung Medical University, No. 100, Shih-Chuang 1st Rd., Sanmin District, Kaohsiung 80708, Taiwan (China); Hon, M.-H. [Department of Materials Science and Engineering, National Cheng Kung University, 1 Ta-Hsueh Road, Tainan 70101, Taiwan (China); Dayeh University, 112 Shan-Jiau Road, Da-Tsuen, Changhua 515, Taiwan (China); Wang, M.-C. [Department of Mechanical Engineering, National Kaohsiung University of Applied Sciences, 415 Chien-kung Road, Kaohsiung 80782, Taiwan (China) and Department of Materials Science and Engineering, National United University, 1 Lien-Da Road, Kung-ching Li, Miao Li 360, Taiwan (China)]. E-mail: mcwang@cc.kuas.edu.tw

2007-05-31

Drug releasing microspheres of poly(3-hydroxybutyric acid)/chitosan (PHB/CTS) with various compositions have been synthesized by both single and double emulsion methods, and collected by a freeze-drying process. In this study, gentamicin was used as an antibacterial medicine coated with PHB. The PHB/CTS microspheres of various compositions prepared by a single emulsion process (SEP) were identified as the major PHB phase together with a minor unknown Phase X by X-ray diffraction (XRD) and FT-IR. However, in the microspheres prepared using a double emulsion process (DEP) the dominant Phase was X and the minor phase was PHB. The size of the PHB/CTS microspheres prepared by SEP increased with the PHB/CTS ratio from 1 {mu}m for 1:1 to 2 {mu}m for 5:1. However, the size of the PHB/CTS microspheres prepared by DEP decreased with the PHB/CTS ratio from 1 {mu}m for 1:1 to 800 nm for 5:1.

Structural and morphological studies on poly(3-hydroxybutyrate acid) (PHB)/chitosan drug releasing microspheres prepared by both single and double emulsion processes

International Nuclear Information System (INIS)

Shih, W.-J.; Chen, Y.-H.; Shih, C.-J.; Hon, M.-H.; Wang, M.-C.

2007-01-01

Drug releasing microspheres of poly(3-hydroxybutyric acid)/chitosan (PHB/CTS) with various compositions have been synthesized by both single and double emulsion methods, and collected by a freeze-drying process. In this study, gentamicin was used as an antibacterial medicine coated with PHB. The PHB/CTS microspheres of various compositions prepared by a single emulsion process (SEP) were identified as the major PHB phase together with a minor unknown Phase X by X-ray diffraction (XRD) and FT-IR. However, in the microspheres prepared using a double emulsion process (DEP) the dominant Phase was X and the minor phase was PHB. The size of the PHB/CTS microspheres prepared by SEP increased with the PHB/CTS ratio from 1 μm for 1:1 to 2 μm for 5:1. However, the size of the PHB/CTS microspheres prepared by DEP decreased with the PHB/CTS ratio from 1 μm for 1:1 to 800 nm for 5:1

PhaM is the physiological activator of poly(3-hydroxybutyrate) (PHB) synthase (PhaC1) in Ralstonia eutropha.

Science.gov (United States)

Pfeiffer, Daniel; Jendrossek, Dieter

2014-01-01

Poly(3-hydroxybutyrate) (PHB) synthase (PhaC1) is the key enzyme of PHB synthesis in Ralstonia eutropha and other PHB-accumulating bacteria and catalyzes the polymerization of 3-hydroxybutyryl-CoA to PHB. Activity assays of R. eutropha PHB synthase are characterized by the presence of lag phases and by low specific activity. It is assumed that the lag phase is caused by the time necessary to convert the inactive PhaC1 monomer into the active dimeric form by an unknown priming process. The lag phase can be reduced by addition of nonionic detergents such as hecameg [6-O-(N-heptyl-carbamoyl)-methyl-α-D-glucopyranoside], which apparently accelerates the formation of PhaC1 dimers. We identified the PHB granule-associated protein (PGAP) PhaM as the natural primer (activator) of PHB synthase activity. PhaM was recently discovered as a novel type of PGAP with multiple functions in PHB metabolism. Addition of PhaM to PHB synthase assays resulted in immediate polymerization of 3HB coenzyme A with high specific activity and without a significant lag phase. The effect of PhaM on (i) PhaC1 activity, (ii) oligomerization of PhaC1, (iii) complex formation with PhaC1, and (iv) PHB granule formation in vitro and in vivo was shown by cross-linking experiments of purified proteins (PhaM, PhaC1) with glutardialdehyde, by size exclusion chromatography, and by fluorescence microscopic detection of de novo-synthesized PHB granules.

Producing a True Lignin Depolymerase for Biobleaching Softwood Kraft Pulp

Energy Technology Data Exchange (ETDEWEB)

Simo Sarkanen

2002-02-04

This project constituted an intensive effort devoted to producing, from the white-rot fungus Tramets Cingulata, a lignin degrading enzyme (lignin depolymerase) that is directly able to biobleach or delignify softwood kraft pulp brownstock. To this end, the solutions in which T. cingulata was grown contained dissolved kraft lignin which fulfilled two functions; it behaved as a lignin deploymerase substrate and it also appeared to act as an inducer of enzyme expression. However, the lignin depolymerase isoenzymes (and other extracellular T. cingulata enzymes) interacted very strongly with both the kraft lignin components and the fungal hypae, so the isolating these proteins from the culture solutions proved to be unexpectedly difficult. Even after extensive experimentation with a variety of protein purification techniques, only one approach appeared to be capable of purifying lignin depolymerases to homogeneity. Unfortunately the procedure was extremely laborious; it involved the iso electric focusing of concentrated buffer-exchanged culture solutions followed by electro-elution of the desired protein bands from the appropriate polyacrylamide gel segments

Osteoconductive bio-based meshes based on Poly(hydroxybutyrate-co-hydroxyvalerate) and poly(butylene adipate-co-terephthalate) blends

International Nuclear Information System (INIS)

Nar, Mangesh; Staufenberg, Gerrit; Yang, Bing; Robertson, Lesli; Patel, Rinkesh H.; Varanasi, Venu G.; D'Souza, Nandika Anne

2014-01-01

Poly(butylene adipate-co-terephthalate) (PBAT) and Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) are biopolymers that have the potential to be used in applications of bone healing. In this study, it is hypothesized that the polymer blend has the combined strength and osteoconductivity to support osteoblast collagen formation. PBAT (PBAT 100), and a blend with 20% PHBV (PBAT 80) were extruded in the form of fibers and then knitted in the form of mesh. These were tested in the warp as well as weft direction for the tensile properties; these showed that the weft direction had higher performance than the warp. The individual fibers were kept in phosphate buffered saline (PBS) over the period of 8 weeks and were tested for the storage and loss modulus using a dynamic mechanical analyser (DMA). The results indicated that mechanical relaxation strength showed a decrease and then an increase. In vitro osteoconductivity studies were done by using differentiating osteoblasts (MC3T3-E1 subclone 4 cells). Environmental Scanning Electron Microscopy (ESEM) showed that pre-soaking the samples in α-MEM for two weeks resulted in cell attachment and growth. X-ray diffraction (XRD) was used to determine the change in structure of polymers due to in vitro degradation for two weeks. Raman spectroscopy showed that all scaffolds supported the formation of a collagenous network over the scaffold surfaces. For a combination of knittable manufacturing, mechanical performance and osteoconductivity, blends offer an effective route. - Highlights: • PBAT and PHBV blend can be knitted to form mesh with good mechanical properties. • PBAT and PHBV blend do not show significant weight loss over a period of 8 weeks in PBS. • Osteoblast cell culture was done on these samples. • They support extracellular matrix and growth and hence are osteoconductive

Osteoconductive bio-based meshes based on Poly(hydroxybutyrate-co-hydroxyvalerate) and poly(butylene adipate-co-terephthalate) blends

Energy Technology Data Exchange (ETDEWEB)

Nar, Mangesh; Staufenberg, Gerrit; Yang, Bing [Department of Material Science and Engineering, University of North Texas, 1155 Union Circle #305310, Denton, TX 76203-5017 (United States); Robertson, Lesli [Department of Fibers, College of Visual Arts and Design, 1155 Union Circle #305100, Denton, TX 76203-5017 (United States); Patel, Rinkesh H. [Department of Biomedical Sciences and Center for Craniofacial Research and Diagnosis, Texas A and M University Baylor College of Dentistry, Dallas, TX 75246 (United States); Varanasi, Venu G., E-mail: vvaranasi@bcd.tamhsc.edu [Department of Biomedical Sciences and Center for Craniofacial Research and Diagnosis, Texas A and M University Baylor College of Dentistry, Dallas, TX 75246 (United States); D' Souza, Nandika Anne, E-mail: nandika.dsouza@unt.edu [Department of Material Science and Engineering, University of North Texas, 1155 Union Circle #305310, Denton, TX 76203-5017 (United States); Department of Mechanical and Energy Engineering, University of North Texas, 1155 Union Circle # 311098, Denton, TX 76203-5017 (United States)

2014-05-01

Poly(butylene adipate-co-terephthalate) (PBAT) and Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) are biopolymers that have the potential to be used in applications of bone healing. In this study, it is hypothesized that the polymer blend has the combined strength and osteoconductivity to support osteoblast collagen formation. PBAT (PBAT 100), and a blend with 20% PHBV (PBAT 80) were extruded in the form of fibers and then knitted in the form of mesh. These were tested in the warp as well as weft direction for the tensile properties; these showed that the weft direction had higher performance than the warp. The individual fibers were kept in phosphate buffered saline (PBS) over the period of 8 weeks and were tested for the storage and loss modulus using a dynamic mechanical analyser (DMA). The results indicated that mechanical relaxation strength showed a decrease and then an increase. In vitro osteoconductivity studies were done by using differentiating osteoblasts (MC3T3-E1 subclone 4 cells). Environmental Scanning Electron Microscopy (ESEM) showed that pre-soaking the samples in α-MEM for two weeks resulted in cell attachment and growth. X-ray diffraction (XRD) was used to determine the change in structure of polymers due to in vitro degradation for two weeks. Raman spectroscopy showed that all scaffolds supported the formation of a collagenous network over the scaffold surfaces. For a combination of knittable manufacturing, mechanical performance and osteoconductivity, blends offer an effective route. - Highlights: • PBAT and PHBV blend can be knitted to form mesh with good mechanical properties. • PBAT and PHBV blend do not show significant weight loss over a period of 8 weeks in PBS. • Osteoblast cell culture was done on these samples. • They support extracellular matrix and growth and hence are osteoconductive.

Efficacy Study of Carrageenan as an Alternative Infused Material (Filler in Poly(3-hydroxybutyrate-co-3-hydroxyvalerate Porous 3D Scaffold

Directory of Open Access Journals (Sweden)

Nor Syamimi Che Johari

2017-01-01

Full Text Available Polymeric porous 3D scaffold plays an important role in culturing mammalian cells as ex vivo model. However, the scaffold used is ineffective due to its structural and cell acceptability weaknesses. Therefore, this research attempts to overcome the weaknesses by using carrageenan from red seaweed Kappaphycus alvarezii as an alternative infused material (filler of poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV porous 3D scaffold. The 3D scaffold was conventionally fabricated using the solvent-casting particulate-leaching (SCPL method. Carrageenan was later infused into 3D porous scaffolds under vacuum pressure and freeze-drying process. Five carrageenan concentrations were prepared and its physicochemical properties such as pH and viscosity were carried out on each concentration to determine the best solutions to produce a new composite 3D structure. The preliminary result shows that carrageenan concentrations of 2, 4, and 6% (w/v were considered the best solutions for the infusion process due to its stable rheology properties. The pH and viscosity profiles of three selected carrageenan solutions were exhibited in the range of 9.00–9.20 and 0.047–1.144 Pa·s, respectively. Moreover, the incorporated carrageenan gel fraction was in the range of 4.30% to 14.95% (w/w which was determined by gravimetric analysis and dye staining method (visual assessment. The well-infused carrageenan 3D scaffold was further characterized based on its internal morphology and degradability study. The vertical cross-sections of the scaffolds revealed homogeneous accumulation of dried gelatinous carrageenan which was covered throughout its pores wall. The degradation rate (K of the carrageenan infused 3D scaffold was between 0.01±1.66 (mg/day and 0.03±3.23 (mg/day. The higher the carrageenan concentration used, the faster the degradation rate occurring (p2 weeks. In conclusion, the usage of carrageenan as a composite material exhibits its great potential to be

Histological and molecular-biological analyses of poly(3-hydroxybutyrate) (PHB) patches for enhancement of bone regeneration.

Science.gov (United States)

Gredes, Tomasz; Gedrange, Tomasz; Hinüber, Claudia; Gelinsky, Michael; Kunert-Keil, Christiane

2015-05-01

Tissue engineered cell-seeded constructs with poly(3)hydroxybutyrate (PHB) induced ectopic bone formation after implantation into the back muscle of rats. The objective of our in vivo study was to evaluate the osteogenic potential of pure PHB patches in surgically created cranial defects. For this, PHB patches were analyzed after implantation in surgically created defects on the cranium of adult male rats. After healing periods of 4, 8 and 12 weeks, the bone tissue specimens containing PHB patches were processed and analyzed histologically as well as molecular-biologically. After 4 weeks, the PHB patches were completely embedded in connective tissue. Eight weeks after PHB insertion, bone regeneration proceeding from bearing bone was found in 50% of all treated animals, whereas all PHB treated cavities showed both bone formation and embedding of the patches in bone 12 weeks after surgery. Furthermore, all slices showed pronounced development of blood vessels. Histomorphometric analysis presented a regenerated bone mean value between 46.4 ± 16.1% and 54.2 ± 19.3% after 4-12 weeks of healing. Caveolin-1 staining in capillary-like structures showed a 1.16-1.38 fold increased expression in PHB treated defects compared to controls. Real-time RT-PCR analyses showed significantly lower expressions of Alpl, Col1a1 and VEGFA in cranium defects after treatment with PHB patches compared to untreated bony defects of the same cranium. Within the limits of the presented animal investigation, it could conclude that the tested PHB patches featured a good biocompatibility and an osteoconductive character. Copyright © 2014 Elsevier GmbH. All rights reserved.

Accumulation of a poly(hydroxyalkanoate) copolymer containing primarily 3-hydroxyvalerate from simple carbohydrate substrates by Rhodococcus sp. NCIMB 40126.

Science.gov (United States)

Haywood, G W; Anderson, A J; Williams, D R; Dawes, E A; Ewing, D F

1991-04-01

A number of taxonomically-related bacteria have been identified which accumulate poly(hydroxyalkanoate) (PHA) copolymers containing primarily 3-hydroxyvalerate (3HV) monomer units from a range of unrelated single carbon sources. One of these, Rhodococcus sp. NCIMB 40126, was further investigated and shown to produce a copolymer containing 75 mol% 3HV and 25 mol% 3-hydroxybutyrate (3HB) from glucose as sole carbon source. Polyesters containing both 3HV and 3HB monomer units, together with 4-hydroxybutyrate (4HB), 5-hydroxyvalerate (5HV) or 3-hydroxyhexanoate (3HHx), were also produced by this organism from certain accumulation substrates. With valeric acid as substrate, almost pure (99 mol% 3HV) poly(3-hydroxyvalerate) was produced. N.m.r. analysis confirmed the composition of these polyesters. The thermal properties and molecular weight of the copolymer produced from glucose were comparable to those of PHB produced by Alcaligenes eutrophus.

Post-processing optimization of electrospun submicron poly(3-hydroxybutyrate) fibers to obtain continuous films of interest in food packaging applications.

Science.gov (United States)

Cherpinski, Adriane; Torres-Giner, Sergio; Cabedo, Luis; Lagaron, Jose M

2017-10-01

Polyhydroxyalkanoates (PHAs) are one of the most researched family of biodegradable polymers based on renewable materials due to their thermoplastic nature and moisture resistance. The present study was targeted to investigate the preparation and characterization of poly(3-hydroxybutyrate) (PHB) films obtained through the electrospinning technique. To convert them into continuous films and then to increase their application interest in packaging, the electrospun fiber mats were subsequently post-processed by different physical treatments. Thus, the effect of annealing time and cooling method on morphology, molecular order, thermal, optical, mechanical, and barrier properties of the electrospun submicron PHB fibers was studied. Annealing at 160°C, well below the homopolyester melting point, was found to be the minimum temperature at which homogeneous transparent films were produced. The film samples that were cooled slowly after annealing showed the lowest permeability to oxygen, water vapor, and limonene. The optimally post-processed electrospun PHB fibers exhibited similar rigidity to conventional compression-molded PHA films, but with enhanced elongation at break and toughness. Films made by this electrospinning technique have many potential applications, such as in the design of barrier layers, adhesive interlayers, and coatings for fiber- and plastic-based food packaging materials.

Recovery and characterization of poly(3-Hydroxybutyric acid ...

African Journals Online (AJOL)

Darshan

Polyhydroxyalkanoates (PHA) are biodegradable polyesters accumulated intracellularly as energy resources by .... 20 min and filtered through Whatman No.1 filter paper and ... heating rate of 20°C air/min from ambient to a final temperature of.

Characterization of a Bio-Based, Biodegradable Class of Copolymers, Poly[(R)-3-Hydroxybutyrate-Co-(R)-3- Hydroxyhexanoate], and Application Development

Science.gov (United States)

Sobieski, Brian

As modern society begins to focus on sustainability and renewable resources there is a growing need for the polymer industry to develop more environmentally friendly materials and practices. Part of this movement can be seen in the use of recycled materials in new products and in the development of bio-based, biodegradable polymers. Bio-based, biodegradable polymers are produced from renewable carbon sources, such as vegetable oils, typically polymerized using fermentation reactions via bacteria, and are able to be consumed by bacteria in landfills to completely convert the polymers to water and CO2. One class of such polymers are poly(hydroxyalkanoate)'s (PHAs), which are chiral, aliphatic polyesters. Within this class of polyesters are poly(hydroxybutyrate) (PHB) and the copolymer poly[(R)-3-hydroxybutyrate- co-(R)-3-hydroxyhexanoate] (PHBHx), which have received extensive study due to their material properties as thermoplastics. Although the properties of PHB have been widely explored, much still remains to be understood about these promising biodegradable polymers. Specifically, PHB and its copolymers exhibit physical gelation in most solvents, yet the origin and mechanism of gelation and the properties of the resulting gel state are unknown. This research effort was primarily focused on investigating the physical gel state of PHBHx. Five goals were laid out and completed: determining the origin of gelation, the mechanism of gelation, the structure of the gel state, the properties of the gel state, and the effects of gelation on electrospun fibers of PHBHx. These goals were achieved through material characterization of the gel state utilizing infrared spectroscopy/two-dimensional correlation spectroscopy, differential scanning calorimetry, X-ray diffraction, scanning electron microscopy, and many other analysis methods. Crystallization of the polymer in solution was found to cause gelation in PHBHx solutions, where the polymer crystals act as tie points forming

PhaC Synthases and PHA Depolymerases: The Enzymes that Produce and Degrade Plastic

Directory of Open Access Journals (Sweden)

Amro A. Amara

2011-12-01

Full Text Available PHAs are a group of intracellular biodegradable polymer produced by (most bacteria under unbalanced growth conditions. A series of enzymes are involved in different PHAs synthesis, however PhaC synthases are responsible for the polymerization step. PHAs are accumulated in bacterial cells from soluble to insoluble form as storage materials inside the inclusion bodies during unbalanced nutrition or to save organisms from reduces equivalents. PHAs are converted again to soluble components by another pathways and enzymes for the degradation process. PHAs depolymerases are the responsible enzymes. This review is designed to give the non-specialists a condense background about PHAs especially for researcher and students in medicinal and pharmaceutical filled. ABSTRAK: PHAs (polyhydroxyalkanoate merupakan sekumpulan polimer terbiodegradasikan intrasel yang dihasilkan oleh (kebanyakan bakteria di bawah keadaan tumbesaran tak seimbang. Satu rangkaian enzim terlibat dalam sistesis PHAs yang berbeza, namun sintesis PhaC bertanggungjawab dalam peringkat pempolimeran. PHAs dikumpulkan dalam sel bakteria dari bentuk larut dan tak larut sebagai bahan simpan di dalam jasad terangkum semasa nutrisi tak seimbang atau untuk menyelamatkan organisma daripada pengurangan tak keseimbangan. PHAs ditukarkan sekali lagi kepada komponen larut dengan cara lain dan enzim lain untuk proses degradasi. PHAs depoly-merases (enzim yang memangkin penguraian makro molekul kepada molekul yang lebih mudah merupakan enzim yang bertanggunjawab. Kajian semula ini direka untuk memberi mereka yang bukan pakar, satu ringkasan tentang PHAs terutamanya penyelidik dan penuntut dalam bidang peubatan dan farmaseutikal.

Effect of water absorption on the mechanical properties of poly(3-hydroxybutyrate)/vegetable fiber composites

Science.gov (United States)

Marinho, Vithória A. D.; Carvalho, Laura H.; Canedo, Eduardo L.

2015-05-01

The present work studies the effect of water absorption on the performance of composites of poly(3-hydroxybutyrate) (PHB) - a fully biodegradable semi-crystalline thermoplastic obtained from renewable resources through low-impact biotechnological process, biocompatible and non-toxic - and vegetable fiber from the fruit (coconut) of babassu palm tree.Water resistance is an important characteristic of structural composites, that may exposed to rain and humid environments. Both water absorption capacity (water solubility in the material) and the rate of water absorption (controlled by the diffusivity of water in the material) are important parameters. However, water absorption per se may not be the most important characteristic, insofar as the performance and applications of the compounds. It is the effect of the water content on the ultimate properties that determine the suitability of the material for applications that involve prolonged exposure to water.PHB/babassu composites with 0-20% load were prepared in an internal mixer. Two different types of babassu fibers having two different article size ranges were compounded with PHB and test specimens molded by compression. The water absorption capacity and the kinetic constant of water absorption were measured in triplicate. Mechanical properties under tension were measured for dry and moist specimens with different amounts of absorbed water.Results indicate that the performance of the composites is comparable to that of the pure matrix. Water absorption capacity increases from 0.7% (pure PHB) to 4% (PHB/20% babassu), but the water diffusivity (4.10□8 cm2/s) was found to be virtually independent of the water absorption level. Water absorption results in moderate drop in elastic modulus (10-30% at saturation, according to fiber content) but has little effect on tensile strength and elongation at break. Fiber type and initial particle size do not have a significant effect on water absorption or mechanical properties.

Lipase catalyzed ultrasonic synthesis of poly-4-hydroxybutyrate-co-6-hydroxyhexanoate.

Science.gov (United States)

Gumel, A M; Annuar, M S M; Chisti, Y

2013-05-01

Four different lipases were compared for ultrasound-mediated synthesis of the biodegradable copolymer poly-4-hydroxybutyrate-co-6-hydroxyhexanoate. The copolymerization was carried out in chloroform. Of the enzymes tested, Novozym 435 exhibited the highest copolymerization rate, in fact the reaction rate was observed to increase with about 26-fold from 30 to 50°C (7.9×10(-3)Ms(-1)), sonic power intensity of 2.6×10(3)Wm(-2) and dissipated energy of 130.4Jml(-1). Copolymerization rates with the Candida antarctica lipase A, Candida rugosa lipase, and Lecitase Ultra™ were lower at 2.4×10(-4), 1.3×10(-4) and 3.5×10(-4)Ms(-1), respectively. The catalytic efficiency depended on the enzyme. The efficiency ranged from 4.15×10(-3)s(-1)M(-1) for Novozym 435-1.48×10(-3)s(-1)M(-1) for C. rugosa lipase. Depending on the enzyme and sonication intensity, the monomer conversion ranged from 8.2% to 48.5%. The sonication power, time and temperature were found to affect the rate of copolymerization. Increasing sonication power intensity from 1.9×10(3) to 4.5×10(3)Wm(-2) resulted in an increased in acoustic pressure (P(a)) from 3.7×10(8) to 5.7×10(8)Nm(-2) almost 2.4-3.7 times greater than the acoustic pressure (1.5×10(8)Nm(-2)) that is required to cause cavitation in water. A corresponding acoustic particle acceleration (a) of 9.6×10(3)-1.5×10(4)ms(-2) was calculated i.e. approximately 984-1500 times greater than under the action of gravity. Copyright © 2012 Elsevier B.V. All rights reserved.

xylA and xylB overexpression as a successful strategy for improving xylose utilization and poly-3-hydroxybutyrate production in Burkholderia sacchari.

Science.gov (United States)

Guamán, Linda P; Oliveira-Filho, Edmar R; Barba-Ostria, Carlos; Gomez, José G C; Taciro, Marilda K; da Silva, Luiziana Ferreira

2018-03-01

Despite the versatility and many advantages of polyhydroxyalkanoates as petroleum-based plastic substitutes, their higher production cost compared to petroleum-based polymers has historically limited their large-scale production. One appealing approach to reducing production costs is to employ less expensive, renewable feedstocks. Xylose, for example is an abundant and inexpensive carbon source derived from hemicellulosic residues abundant in agro-industrial waste (sugarcane bagasse hemicellulosic hydrolysates). In this work, the production of poly-3-hydroxybutyrate P(3HB) from xylose was studied to develop technologies for conversion of agro-industrial waste into high-value chemicals and biopolymers. Specifically, this work elucidates the organization of the xylose assimilation operon of Burkholderia sacchari, a non-model bacterium with high capacity for P(3HB) accumulation. Overexpression of endogenous xylose isomerase and xylulokinase genes was successfully assessed, improving both specific growth rate and P(3HB) production. Compared to control strain (harboring pBBR1MCS-2), xylose utilization in the engineered strain was substantially improved with 25% increase in specific growth rate, 34% increase in P(3HB) production, and the highest P(3HB) yield from xylose reported to date for B. sacchari (Y P3HB/Xil  = 0.35 g/g). This study highlights that xylA and xylB overexpression is an effective strategy to improve xylose utilization and P(3HB) production in B. sacchari.

The ntrB and ntrC Genes Are Involved in the Regulation of Poly-3-Hydroxybutyrate Biosynthesis by Ammonia in Azospirillum brasilense Sp7

Science.gov (United States)

Sun, Jun; Peng, Xuan; Van Impe, Jan; Vanderleyden, Jos

2000-01-01

Azospirillum brasilense Sp7 and its ntrA (rpoN), ntrBC, and ntrC mutants have been evaluated for their capabilities of poly-3-hydroxybutyrate (PHB) accumulation in media with high and low ammonia concentrations. It was observed that the ntrBC and ntrC mutants can produce PHB in both low- and high-C/N-ratio media, while no significant PHB production was observed for the wild type or the ntrA mutant in low-C/N-ratio media. Further investigation by fermentation analysis indicated that the ntrBC and ntrC mutants were able to grow and accumulate PHB simultaneously in the presence of a high concentration of ammonia in the medium, while little PHB was produced in the wild type and ntrA (rpoN) mutant during active growth phase. These results provide the first genetic evidence that the ntrB and ntrC genes are involved in the regulation of PHB synthesis by ammonia in A. brasilense Sp7. PMID:10618211

Kajian Biodegradasi Filem Plastik Campuran Polistiren Dengan Poli (3-Hidroksibutirat) Dalam Tanah

OpenAIRE

Octaviani, Melzi; Zaini, Erizal; Djamaan, Akmal

2013-01-01

The degradation of a plastic film containing polymer synthetic polystyrene (PS) and biopolymer poly(3-hydroxybutyrate) [P(3HB)] in soil has been carried out based on soil burial test. Result showed that the amount of P(3HB) in plastic film containing PS and P(3HB) influence the degradation rate of plastic film. Rate degradation measured use linear regression equality and parameter is indicated by the slope (k), degradation time 50% (t 50%), and degradation time 95% (t 95%). Degradation plasti...

Gamma Irradiation Effect on Biodegradable Poly (Hydroxybutyrate) Studied by Positron Annihilation Technique

International Nuclear Information System (INIS)

Abdel-Hady, E.E.; Mohamed, S.S.

2010-01-01

-Bacterial polyesters have attracted much attention as biodegradable polymers. An ecofriendly alternative to this biodegradable material is poly-3-hydroxybutyrate (PHB) which has attracted industrial attention as an environmentally degradable plastic for a wide range of medical applications. Free volume holes in polymers play a crucial role in determining its physical properties. The Positron Annihilation Lifetime (PAL) technique has been established as a powerful probe for microstructures of polymers, in particular, angstrom-sized free volume holes. The PHB samples were irradiated using 60 Co source at room temperature with doss ranging from 5 to 300 kGy. The PAL spectra for all the samples have been measured at room temperature as a function of gamma-irradiation dose. The free volume hole size decreases with increasing the irradiation dose up to 25 kGy followed by slowly increases up to 200 kGy, then decreases at higher doses. On the other hand, the free volume content decreases with increasing the gamma-irradiation dose which is due to the increase of the degree of crystallinity. The variations in the free volume with the irradiation dose will be discussed in the frame of free volume model. A correlation between the macroscopic mechanical properties Hv and positron annihilation parameters has been done

A Long-Acting BMP-2 Release System Based on Poly(3-hydroxybutyrate) Nanoparticles Modified by Amphiphilic Phospholipid for Osteogenic Differentiation

Science.gov (United States)

Peng, Xiaochun; Chen, Yunsu; Li, Yamin; Wang, Yiming

2016-01-01

We explored a novel poly(3-hydroxybutyrate) (PHB) nanoparticle loaded with hydrophilic recombinant human BMP-2 with amphiphilic phospholipid (BPC-PHB NP) for a rapid-acting and long-acting delivery system of BMP-2 for osteogenic differentiation. The BPC-PHB NPs were prepared by a solvent evaporation method and showed a spherical particle with a mean particle size of 253.4 nm, mean zeta potential of −22.42 mV, and high entrapment efficiency of 77.18%, respectively. For BPC-PHB NPs, a short initial burst release of BMP-2 from NPs in 24 h was found and it has steadily risen to reach about 80% in 20 days for in vitro test. BPC-PHB NPs significantly reduced the burst release of BMP-2, as compared to that of PHB NPs loading BMP-2 without PL (B-PHB NPs). BPC-PHB NPs maintained the content of BMP-2 for a long-term osteogenic differentiation. The OCT-1 cells with BPC-PHB NPs have high ALP activity in comparison with others. The gene markers for osteogenic differentiation were significantly upregulated for sample with BPC-PHB NPs, implying that BPC-PHB NPs can be used as a rapid-acting and long-acting BMP-2 delivery system for osteogenic differentiation. PMID:27379249

Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

OpenAIRE

Kadouri, Daniel; Burdman, Saul; Jurkevitch, Edouard; Okon, Yaacov

2002-01-01

Like many other prokaryotes, rhizobacteria of the genus Azospirillum produce high levels of poly(β-hydroxybutyrate) (PHB) under suboptimal growth conditions. Utilization of PHB by bacteria under stress has been proposed as a mechanism that favors their compatible establishment in competitive environments, thus showing great potential for the improvement of bacterial inoculants for plants and soils. The three genes that are considered to be essential in the PHB biosynthetic pathway, phbA (β-ke...

Antimicrobial and cell viability measurement of bovine serum albumin capped silver nanoparticles (Ag/BSA) loaded collagen immobilized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) film.

Science.gov (United States)

Bakare, Rotimi; Hawthrone, Samantha; Vails, Carmen; Gugssa, Ayele; Karim, Alamgir; Stubbs, John; Raghavan, Dharmaraj

2016-03-01

Bacterial infection of orthopedic devices has been a major concern in joint replacement procedures. Therefore, this study is aimed at formulating collagen immobilized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) film loaded with bovine serum albumin capped silver nanoparticles (Ag/BSA NPs) to inhibit bacterial growth while retaining/promoting osteoblast cells viability. The nanoparticles loaded collagen immobilized PHBV film was characterized for its composition by X-ray Photoelectron Spectroscopy and Anodic Stripping Voltammetry. The extent of loading of Ag/BSA NPs on collagen immobilized PHBV film was found to depend on the chemistry of the functionalized PHBV film and the concentration of Ag/BSA NPs solution used for loading nanoparticles. Our results showed that more Ag/BSA NPs were loaded on higher molecular weight collagen immobilized PHEMA-g-PHBV film. Maximum loading of Ag/BSA NPs on collagen immobilized PHBV film was observed when 16ppm solution was used for adsorption studies. Colony forming unit and optical density measurements showed broad antimicrobial activity towards Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa at significantly lower concentration i.e., 0.19 and 0.31μg/disc, compared to gentamicin and sulfamethoxazole trimethoprim while MTT assay showed that released nanoparticles from Ag/BSA NPs loaded collagen immobilized PHBV film has no impact on MCTC3-E1 cells viability. Copyright © 2015 Elsevier Inc. All rights reserved.

Effect of co-substrate on production of poly-β- hydroxybutyrate (PHB and copolymer PHBV from newly identified mutant Rhodobacter sphaeroides U7 cultivated under aerobic-dark condition

Directory of Open Access Journals (Sweden)

Kemarajt Kemavongse

2007-07-01

Full Text Available Photosynthetic bacterial mutant strain U7 was identified using both classical and molecular (16S rDNA techniques to be Rhodobacter sphaeroides. The glutamate-acetate (GA medium containing sodium acetate and sodium glutamate as carbon and nitrogen sources was used for production of poly-β-hydroxybutyrate (PHB from R. sphaeroides U7 cultivated under aerobic-dark condition (200 rpm at 37oC. Effect of auxiliary carbon sources (propionate and valerate and concentrations (molar ratio of 40/0, 40/20, 40/40 and 40/80 on copolymer production were studied. Both combinations of acetate with valerate and acetate with propionate were found to induce the accumulation of poly-β-hydroxybutyrate-co-β-hydroxyvalerate (PHBV within the cell. Acetate with propionate in the molar ratio of 40/40 gave the highest poly-β-hydroxyalkanoates (PHA content (77.68%, followed by acetate with valerate at the same molar ratio (77.42%. Although their polymer contents were similar, the presence of 40 mM valerate gave more than 4 times higher hydroxyvalerate (HV fraction (84.77% than in the presence of 40 mM propionate (19.12% HV fraction.

Poly (3-hydroxybutyrate-co-15 mol% 3hydroxyhexanoate)/ZnO nanocomposites by solvent casting method: a study of optical, surface, and thermal properties

Science.gov (United States)

Vishnu Chandar, J.; Shanmugan, S.; Mutharasu, D.; Azlan, A. A.

2017-01-01

Biopolymer nanocomposites are gaining interest due to their biodegradable and biocompatible nature, with exceptional properties which are superior to those of pure polymer and that can be used as a replacement for petroleum-based plastics. Pure poly (3-hydroxybutyrate-co-15 mol% 3hydroxyhexanoate) [P(3HB-co-15 mol% 3HHx)] and ZnO nanoparticles (NPs) reinforced P(3HB-co-15 mol% 3HHx) composite films were prepared and their morphological, optical, and thermal analyses were carried out to study the effect of ZnO NPs on P(3HB-co-15 mol% 3HHx) copolymer. From optical analysis, a strong absorbance peak at ˜358 nm with high intensity for 5%ZnO NPs reinforced P(3HB-co-15 mol% 3HHx) composite sample was observed. Nano-sized particles and their increased trend with increased ZnO NPs concentration were confirmed by UV-Vis spectral analysis. In addition to that, ZnO NPs reduced the reflectance behaviour of P(3HB-co-15 mol% 3HHx) copolymer in the UV region. Field emission scanning electron microscopy analysis evidenced that the pore numbers and their size in the composite matrix was decreased with increased ZnO NPs concentration. Improved thermal stability and melting temperature of the copolymer matrix was confirmed by thermogravimetric analysis and differential scanning calorimetry analysis, respectively. ZnO NPs acted as a retarding agent and showed a low degree of crystallinity (X c ) and enthalpy of melting (ΔH M ). From these observations, ZnO NPs reinforced P(3HB-co-15 mol% 3HHx) composites can be integrated with suitable resin and can be used as an LED encapsulant to block UV rays. They can also be used as a UV blocking coating material in the glass, plastic, and textile industries, for NIR shielding and food packaging applications.

Effect of Organo-Modified Nanoclay on the Thermal and Bulk Structural Properties of Poly(3-hydroxybutyrate-Epoxidized Natural Rubber Blends: Formation of Multi-Components Biobased Nanohybrids

Directory of Open Access Journals (Sweden)

Ali Salehabadi

2014-06-01

Full Text Available Multi-component nanohybrids comprising of organo-modified montmorillonite (MMT and immiscible biopolymer blends of poly(3-hydroxybutyrate (PHB and epoxidized natural rubber (ENR-50 were prepared by solvent casting technique. The one and three dimensional morphology of PHB/ENR-50/MMT systems were studied using Polarizing Optical Microscopy (POM and Scanning Electron Microscopy (SEM. Differential scanning calorimetry (DSC technique was used to evaluate the thermal properties of the nanohybrids. The melting temperature (Tm and enthalpy of melting (ΔHm of PHB decrease with respect to the increase in ENR-50 as well as MMT content. The non-isothermal decomposition of the nanohybrids was studied using thermogravimetric (TG-DTG analysis. FTIR-ATR spectra supported ring opening of the epoxide group via reaction with carboxyl group of PHB and amines of organic modifier. The reaction mechanism towards the formation of the nanohybrids is proposed.

In vitro biocompatibility of 45S5 Bioglass-derived glass-ceramic scaffolds coated with poly(3-hydroxybutyrate).

Science.gov (United States)

Bretcanu, Oana; Misra, Superb; Roy, Ipsita; Renghini, Chiara; Fiori, Fabrizio; Boccaccini, Aldo R; Salih, Vehid

2009-02-01

The aim of this work was to study the in vitro biocompatibility of glass-ceramic scaffolds based on 45S5 Bioglass, using a human osteosarcoma cell line (HOS-TE85). The highly porous scaffolds were produced by the foam replication technique. Two different types of scaffolds with different porosities were analysed. They were coated with a biodegradable polymer, poly(3-hydroxybutyrate) (P(3HB)). The scaffold bioactivity was evaluated by soaking in a simulated body fluid (SBF) for different durations. Compression strength tests were performed before and after immersion in SBF. These experiments showed that the scaffolds are highly bioactive, as after a few days of immersion in SBF a hydroxyapatite-like layer was formed on the scaffold's surface. It was also observed that P(3HB)-coated samples exhibited higher values of compression strength than uncoated samples. Biocompatibility assessment was carried out by qualitative evaluation of cell morphology after different culture periods, using scanning electron microscopy, while cell proliferation was determined by using the AlamarBlue assay. Alkaline phosphatase (ALP) and osteocalcin (OC) assays were used as quantitative in vitro indicators of osteoblast function. Two different types of medium were used for ALP and OC tests: normal supplemented medium and osteogenic medium. HOS cells were seeded and cultured onto the scaffolds for up to 2 weeks. The AlamarBlue assay showed that cells were able to proliferate and grow on the scaffold surface. After 7 days in culture, the P(3HB)-coated samples had a higher number of cells on their surfaces than the uncoated samples. Regarding ALP- and OC-specific activity, no significant differences were found between samples with different pore sizes. All scaffolds containing osteogenic medium seemed to have a slightly higher level of ALP and OC concentration. These experiments confirmed that Bioglass/P(3HB) scaffolds have potential as osteoconductive tissue engineering substrates for

Ventral hernia repair with poly-4-hydroxybutyrate mesh.

Science.gov (United States)

Plymale, Margaret A; Davenport, Daniel L; Dugan, Adam; Zachem, Amanda; Roth, John Scott

2018-04-01

Biomaterial research has made available a biologically derived fully resorbable poly-4-hydroxybutyrate (P4HB) mesh for use in ventral and incisional hernia repair (VIHR). This study evaluates outcomes of patients undergoing VIHR with P4HB mesh. An IRB-approved prospective pilot study was conducted to assess clinical and quality of life (QOL) outcomes for patients undergoing VIHR with P4HB mesh. Perioperative characteristics were defined. Clinical outcomes, employment status, QOL using 12-item short form survey (SF-12), and pain assessments were followed for 24 months postoperatively. 31 patients underwent VIHR with bioresorbable mesh via a Rives-Stoppa approach with retrorectus mesh placement. The median patient age was 52 years, median body mass index was 33 kg/m 2 , and just over half of the patients were female. Surgical site occurrences occurred in 19% of patients, most of which were seroma. Hernia recurrence rate was 0% (median follow-up = 414 days). Patients had significantly improved QOL at 24 months compared to baseline for SF-12 physical component summary and role emotional (p < 0.05). Ventral hernia repair with P4HB bioresorbable mesh results in favorable outcomes. Early hernia recurrence was not identified among the patient cohort. Quality of life improvements were noted at 24 months versus baseline for this cohort of patients with bioresorbable mesh. Use of P4HB mesh for ventral hernia repair was found to be feasible in this patient population. (ClinicalTrials.gov Identifier: NCT01863030).

Use of NAD(P)H fluorescence measurement for on-line monitoring of metabolic state of Azohydromonas australica in poly(3-hydroxybutyrate) production.

Science.gov (United States)

Gahlawat, Geeta; Srivastava, Ashok K

2013-02-01

Culture fluorescence measurement is an indirect and non-invasive method of biomass estimation to assess the metabolic state of the microorganism in a fermentation process. In the present investigation, NAD(P)H fluorescence has been used for on-line in situ characterization of metabolic changes occurring during different phases of batch cultivation of Azohydromonas australica in growth associated poly(3-hydroxybutyrate) or PHB production. A linear correlation between biomass concentration and net NAD(P)H fluorescence was obtained during early log phase (3-12 h) and late log phase (24-39 h) of PHB fermentation. After 12 h (mid log phase) cultivation PHB accumulation shot up and a drop in culture fluorescence was observed which synchronously exhibited continuous utilization of NAD(P)H for the synthesis of biomass and PHB formation simultaneously. A decrease in the observed net fluorescence value was observed again towards the end of fermentation (at 39 h) which corresponded very well with the culture starvation and substrate depletion towards the end of cultivation inside the bioreactor. It was therefore concluded that NAD(P)H fluorescence measurements could be used for indication of the time of fresh nutrient (substrate) feed during substrate limitation to further enhance the PHB production.

Reducible poly(amido ethylenimine)s designed for triggered intracellular gene delivery

NARCIS (Netherlands)

Christensen, Lane V.; Christensen, L.; Chang, Chien-Wen; Kim, Won Jong; Kim, Sung Wan; Zhong, Zhiyuan; Lin, C.; Engbersen, Johannes F.J.; Feijen, Jan

2006-01-01

Poly(amido ethylenimine) polymers, a new type of peptidomimetic polymer, containing multiple disulfide bonds (SS-PAEIs) designed to degrade after delivery of plasmid DNA (pDNA) into the cell were synthesized and investigated as new carriers for triggered intracellular gene delivery. More

Biocomposite scaffolds based on electrospun poly(3-hydroxybutyrate) nanofibers and electrosprayed hydroxyapatite nanoparticles for bone tissue engineering applications

International Nuclear Information System (INIS)

Ramier, Julien; Bouderlique, Thibault; Stoilova, Olya; Manolova, Nevena; Rashkov, Iliya; Langlois, Valérie; Renard, Estelle; Albanese, Patricia; Grande, Daniel

2014-01-01

The electrospinning technique combined with the electrospraying process provides a straightforward and versatile approach for the fabrication of novel nanofibrous biocomposite scaffolds with structural, mechanical, and biological properties potentially suitable for bone tissue regeneration. In this comparative investigation, three types of poly(3-hydroxybutyrate) (PHB)-based scaffolds were engineered: (i) PHB mats by electrospinning of a PHB solution, (ii) mats of PHB/hydroxyapatite nanoparticle (nHA) blends by electrospinning of a mixed solution containing PHB and nHAs, and (iii) mats constituted of PHB nanofibers and nHAs by simultaneous electrospinning of a PHB solution and electrospraying of a nHA dispersion. Scaffolds based on PHB/nHA blends displayed improved mechanical properties compared to those of neat PHB mats, due to the incorporation of nHAs within the fibers. The electrospinning/electrospraying approach afforded biocomposite scaffolds with lower mechanical properties, due to their higher porosity, but they displayed slightly better biological properties. In the latter case, the bioceramic, i.e. nHAs, largely covered the fiber surface, thus allowing for a direct exposure to cells. The 21 day-monitoring through the use of MTS assays and SEM analyses demonstrated that human mesenchymal stromal cells (hMSCs) remained viable on PHB/nHA biocomposite scaffolds and proliferated continuously until reaching confluence. - Highlights: • Three different types of PHB-based scaffolds are engineered and thoroughly investigated. • The combination of electrospinning and electrospraying affords original nanofibrous biocomposite scaffolds. • PHB-based scaffolds show a strong capability of supporting viable cell development for 21 days

Biocomposite scaffolds based on electrospun poly(3-hydroxybutyrate) nanofibers and electrosprayed hydroxyapatite nanoparticles for bone tissue engineering applications

Energy Technology Data Exchange (ETDEWEB)

Ramier, Julien [Institut de Chimie et des Matériaux Paris-Est, UMR 7182 CNRS, Université Paris-Est Créteil, 2, rue Henri Dunant, 94320 Thiais (France); Bouderlique, Thibault [Laboratoire “Croissance, Réparation et Régénération Tissulaires”, EAC 7149 CNRS, Université Paris-Est Créteil, 61, avenue du Général de Gaulle, 94010 Créteil (France); Stoilova, Olya; Manolova, Nevena; Rashkov, Iliya [Laboratory of Bioactive Polymers, Institute of Polymers, Bulgarian Academy of Sciences, Acad. G. Bonchev St., bl. 103A, BG-1113 Sofia (Bulgaria); Langlois, Valérie; Renard, Estelle [Institut de Chimie et des Matériaux Paris-Est, UMR 7182 CNRS, Université Paris-Est Créteil, 2, rue Henri Dunant, 94320 Thiais (France); Albanese, Patricia [Laboratory of Bioactive Polymers, Institute of Polymers, Bulgarian Academy of Sciences, Acad. G. Bonchev St., bl. 103A, BG-1113 Sofia (Bulgaria); Grande, Daniel, E-mail: grande@icmpe.cnrs.fr [Institut de Chimie et des Matériaux Paris-Est, UMR 7182 CNRS, Université Paris-Est Créteil, 2, rue Henri Dunant, 94320 Thiais (France)

2014-05-01

The electrospinning technique combined with the electrospraying process provides a straightforward and versatile approach for the fabrication of novel nanofibrous biocomposite scaffolds with structural, mechanical, and biological properties potentially suitable for bone tissue regeneration. In this comparative investigation, three types of poly(3-hydroxybutyrate) (PHB)-based scaffolds were engineered: (i) PHB mats by electrospinning of a PHB solution, (ii) mats of PHB/hydroxyapatite nanoparticle (nHA) blends by electrospinning of a mixed solution containing PHB and nHAs, and (iii) mats constituted of PHB nanofibers and nHAs by simultaneous electrospinning of a PHB solution and electrospraying of a nHA dispersion. Scaffolds based on PHB/nHA blends displayed improved mechanical properties compared to those of neat PHB mats, due to the incorporation of nHAs within the fibers. The electrospinning/electrospraying approach afforded biocomposite scaffolds with lower mechanical properties, due to their higher porosity, but they displayed slightly better biological properties. In the latter case, the bioceramic, i.e. nHAs, largely covered the fiber surface, thus allowing for a direct exposure to cells. The 21 day-monitoring through the use of MTS assays and SEM analyses demonstrated that human mesenchymal stromal cells (hMSCs) remained viable on PHB/nHA biocomposite scaffolds and proliferated continuously until reaching confluence. - Highlights: • Three different types of PHB-based scaffolds are engineered and thoroughly investigated. • The combination of electrospinning and electrospraying affords original nanofibrous biocomposite scaffolds. • PHB-based scaffolds show a strong capability of supporting viable cell development for 21 days.

Investigation of miscibility of p(3hydroxybutyrate-co-3hydroxyhexanoate) and epoxidized natural rubber blends

Science.gov (United States)

Akram, Faridah; Chan, Chin Han; Natarajan, Valliyappan David

2015-08-01

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate [P(3HB-co-3HHx)] produced by C. necator PHB-4 harboring phaCcs from crude palm kernel oil with 21 mol% of 3-hydroxyhexanoate and epoxidized natural rubber with 25 mol% of epoxy content (ENR-25) were used to study the miscibility of the blends by attenuated total reflection-Fourier transform infrared (ATR-FTIR) and differential scanning calorimetry (DSC). The polymers used were purified and the blends were prepared by solution casting method. Nuclear magnetic resonance (NMR) spectra confirm the purity and molecular structures of P(3HB-co-3HHx) and ENR-25. FTIR spectra for different compositions of P(3HB-co-3HHx) and ENR-25 blends show absorbance change of the absorbance bands but with no significant shifting of the absorbance bands as the P(3HB-co-3HHx) content decreases, which shows that there is no intermolecular interaction between the parent polymer blends. On top of that, there are two Tgs present for the blends and both remain constant for different compositions which corresponds to the Tgs of the parent polymers. This indicates that the blends are immiscible.

Functionalized linear poly(amidoamine)s are efficient vectors for intracellular protein delivery

NARCIS (Netherlands)

Coué, G.M.J.P.C.; Engbersen, Johannes F.J.

2011-01-01

An effective intracellular protein delivery system was developed based on functionalized linear poly(amidoamine)s (PAAs) that form self-assembled cationic nanocomplexes with oppositely charged proteins. Three differently functionalized PAAs were synthesized, two of these having repetitive disulfide

Novel poly(hydroxyalkanoates)-based composites containing Bioglass® and calcium sulfate for bone tissue engineering

International Nuclear Information System (INIS)

García-García, J M; Boccaccini, A R; Garrido, L; Quijada-Garrido, I; Kaschta, J; Schubert, D W

2012-01-01

Three different poly(hydroxyalkanoates) (PHAs), copolymers of poly(3-hydroxybutyrate) (P3HB), have been used to make composites using two different fillers, bioactive glass (type 45S5 Bioglass®) and calcium sulfate dihydrate. The PHAs used were poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [PHBHV] and two copolymers of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [PHBHHx]. The aim of the study was the fabrication and characterization of the new composites and the assessment of the influence of the particular filler combination on the physical properties and bioactivity of the films. The thermal behaviour was studied using differential scanning calorimetry while mechanical properties were evaluated using dynamic mechanic thermal analysis and tensile strength test. The mechanical and thermal properties were affected by particles addition. The distribution of the particles in the polymer matrix, observed by scanning electron microscopy, was directly related to the mechanical properties. The surface characteristics were investigated by contact angle measurements and Raman spectroscopy. The extent of formation of hydroxyapatite (HA) upon immersion in simulated body fluid (SBF) depended on the polymer used, the amount of fillers employed and the time of immersion in SBF. Bioactivity was enhanced in the composites with a rise of hydrophilicity. The HA formation was controllable with time in the case of PHBHHx composites. (paper)

Intracellular siRNA delivery dynamics of integrin-targeted, PEGylated chitosan-poly(ethylene imine) hybrid nanoparticles

DEFF Research Database (Denmark)

Ragelle, Héloïse; Colombo, Stefano; Pourcelle, Vincent

2015-01-01

chitosan-poly(ethylene imine) hybrid nanoparticles. The amount of intracellular siRNA delivered by αvβ3-targeted versus non-targeted nanoparticles was quantified in the human non-small cell lung carcinoma cell line H1299 expressing enhanced green fluorescent protein (EGFP) using a stem-loop reverse...... that these nanoparticles might end up in late endosomes or lysosomes without releasing their cargo to the cell cytoplasm. Thus, the silencing efficiency of the chitosan-based nanoparticles is strongly dependent on the uptake and the intracellular trafficking in H1299 EGFP cells, which is critical information towards...

Bioreducible poly(amidoamine)s with charge-reversel properties for intracellular protein delivery

NARCIS (Netherlands)

Coué, G.M.J.P.C.; Engbersen, Johannes F.J.; Hennink, W.E.; Engbersen, J.F.J.

2010-01-01

An effective intracellular protein delivery system was developed using bioreducible disulfide-containing poly(amidoamine)s with negatively charged citraconic side groups that can give charge-reversal upon pH decrease. These water-soluble and linear polymers efficiently self-assemble with proteins

Produção de poli(3-hidroxibutirato por Cupriavidus Necator em meio hidrolisado de amido de arroz com suplementação de óleo de soja em diferentes temperaturas Production of poly(3-hydroxybutyrate by Cupriavidus Necator in hydrolyzed rice starch medium with soybean oil supplementation at different temperatures

Directory of Open Access Journals (Sweden)

Francieli Dalcanton

2010-01-01

Full Text Available Poly (3-hydroxybutyrate (P(3HB is a biopolymer, completely biodegradable, which has similar properties to fuel-based polymers. However to make it economically competitive it is necessary the study of cheap sources of substrate. The influence of hydrolyzed rice starch supplemented with soybean oil at different temperatures (30, 35 and 40 °C was studied in the production of P(3HB by C. necator. The percentage of P(3HB produced in the cultures at 30, 35 °C was 30, 39% and 35, 43% without and with supplementation of oil, respectively. The culture at 40 °C showed no production phase due to a possible oxygen limitation. These results demonstrate that hydrolyzed rice starch supplemented with soybean oil increases the yield of P(3HB and temperature of 35 ºC is the most favorable for biopolymer production.

Effects of dietary poly-β-hydroxybutyrate (PHB) on microbiota composition and the mTOR signaling pathway in the intestines of litopenaeus vannamei.

Science.gov (United States)

Duan, Yafei; Zhang, Yue; Dong, Hongbiao; Wang, Yun; Zhang, Jiasong

2017-12-01

Poly-β-hydroxybutyrate (PHB) is a natural polymer of the short chain fatty acid β-hydroxybutyrate, which acts as a microbial control agent. The mammalian target of the rapamycin (mTOR) signaling pathway plays a crucial role in intestine inflammation and epithelial morphogenesis. In this study, we examined the composition of intestine microbiota, and mTOR signaling-related gene expression in Pacific white shrimp Litopenaeus vannamei fed diets containing different levels of PHB: 0% (Control), 1% (PHB1), 3% (PHB3), and 5% (PHB5) (w/w) for 35 days. High-throughput sequencing analysis revealed that dietary PHB altered the composition and diversity of intestine microbiota, and that the microbiota diversity decreased with the increasing doses of PHB. Specifically, dietary PHB increased the relative abundance of Proteobacteria and Tenericutes in the PHB1 and PHB5 groups, respectively, and increased that of Gammaproteobacteria in the three PHB groups. Alternatively, PHB decreased Alphaproteobacteria in the PHB3 and PHB5 groups. At the genus level, dietary PHB increased the abundance of beneficial bacteria, such as Bacillus, Lactobacillus, Lactococcus, Clostridium, and Bdellovibrio. The relative mRNA expression levels of the mTOR signaling-related genes TOR, 4E-BP, eIF4E1α, and eIF4E2 all increased in the three PHB treatment groups. These results revealed that dietary PHB supplementation had a beneficial effect on intestine health of L. vannamei by modulating the composition of intestine microbiota and activating mTOR signaling.

Protective effects of exogenous β-hydroxybutyrate on paraquat toxicity in rat kidney

Energy Technology Data Exchange (ETDEWEB)

Wei, Teng; Tian, Wulin; Liu, Fangning; Xie, Guanghong, E-mail: xiegh@jlu.edu.cn

2014-05-16

Highlights: • β-Hydroxybutyrate inhibits paraquat-induced toxicity in rat kidney. • β-Hydroxybutyrate inhibits lipid peroxidation and caspase-mediated apoptosis. • β-Hydroxybutyrate increases the activities of SOD and CAT. • The study describes a novel finding for the renoprotective ability of β-hydroxybutyrate. - Abstract: In this study, we demonstrated the protective effects of β-hydroxybutyrate (β-HB) against paraquat (PQ)-induced kidney injury and elucidated the underlying molecular mechanisms. By histological examination and renal dysfunction specific markers (serum BUN and creatinine) assay, β-HB could protect the PQ-induced kidney injury in rat. PQ-induced kidney injury is associated with oxidative stress, which was measured by increased lipid peroxidation (MDA) and decreased intracellular anti-oxidative abilities (SOD, CAT and GSH). β-HB pretreatment significantly attenuated that. Caspase-mediated apoptosis pathway contributed importantly to PQ toxicity, as revealed by the activation of caspase-9/-3, cleavage of PARP, and regulation of Bcl-2 and Bax, which were also effectively blocked by β-HB. Moreover, treatment of PQ strongly decreased the nuclear Nrf2 levels. However, pre-treatment with β-HB effectively suppressed this action of PQ. This may imply the important role of β-HB on Nrf2 pathway. Taken together, this study provides a novel finding that β-HB has a renoprotective ability against paraquat-induced kidney injury.

Effects of ethyl cellulose on the crystallization and mechanical properties of poly(β-hydroxybutyrate).

Science.gov (United States)

Chen, Jianxiang; Wu, Defeng; Pan, Keren

2016-07-01

Ethyl cellulose (EC) was blended with poly(β-hydroxybutyrate) (PHB), aiming at controlling crystallization and mechanical properties of PHB. The obtained PHB/EC blend is an immiscible system, and the discrete EC phase behaves dual characteristics in the PHB matrix, as the viscoelastic droplets during processing, and as the rigid filler particles during shear flow. This is confirmed by the rheological tests. The presence of EC domains acts as the tackifier, sharply increasing system viscosity from 1000Pas to 5000Pas, and as a result, has large influence on the spherulite morphology of PHB and its crystallization kinetics. The PHB spherulite growth rate reduces in the presence of inert EC, accompanied by decreased degree of crystallinity and reduced lamella defects. These affect the mechanical properties of PHB strongly, together with reinforcing effect of EC itself. At the lower content level, EC can act as both reinforcement and toughener. The presence of 1wt% EC enhances the tensile strength of PHB by about 22%, from 27.5MPa to 33.3MPa, accompanied by 15% increase of impact strength. This work provide an easy way to control the structure and properties of PHB using EC. Copyright © 2016 Elsevier B.V. All rights reserved.

Analysis of Poly-β-Hydroxybutyrate in Rhizobium japonicum Bacteroids by Ion-Exclusion High-Pressure Liquid Chromatography and UV Detection †

Science.gov (United States)

Karr, Dale B.; Waters, James K.; Emerich, David W.

1983-01-01

Ion-exclusion high-pressure liquid chromatography (HPLC) was used to measure poly-β-hydroxybutyrate (PHB) in Rhizobium japonicum bacteroids. The products in the acid digest of PHB-containing material were fractionated by HPLC on Aminex HPX-87H ion-exclusion resin for organic acid analysis. Crotonic acid formed from PHB during acid digestion was detected by its intense absorbance at 210 nm. The Aminex-HPLC method provides a rapid and simple chromatographic technique for routine analysis of organic acids. Results of PHB analysis by Aminex-HPLC were confirmed by gas chromatography and spectrophotometric analysis. PMID:16346443

In vitro blood compatibility of poly (hydroxybutyrate-co-hydroxyhexanoate) and the influence of surface modification by alkali treatment

International Nuclear Information System (INIS)

Shen Feng; Zhang Erlin; Wei Zunjie

2010-01-01

In vitro blood compatibility of poly (hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx) was evaluated in comparison with poly (L-lactic acid) (PLLA) by a haemolysis assay, in vitro platelet adhesion test and coagulation measurements including plasma recalcification time (PRT), plasma prothrombin time (PT) and kinetic clotting time. The results showed that PHBHHx exhibited better blood compatibility than PLLA. Furthermore, PHBHHx film was modified by NaOH treatment to improve the surface hydrophilic property and the influence of the surface modification on the blood compatibility was investigated. Surface properties including hydrophilic property, surface appearance and functional groups were characterized by water contact angle measurement, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The results showed that the hydrophilic property of PHBHHx film was obviously improved by the NaOH treatment. It was also shown that the NaOH treatment could significantly enhance the blood compatibility of PHBHHx by prolonging PRT, PT, and kinetic clotting time and decreasing platelet activation. It is thought that the improvement in the hydrophilic property mainly contributes to the enhancement of blood compatibility.

Investigation of miscibility of p(3hydroxybutyrate-co-3hydroxyhexanoate) and epoxidized natural rubber blends

Energy Technology Data Exchange (ETDEWEB)

Akram, Faridah; Chan, Chin Han; Natarajan, Valliyappan David [Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, 40450 Selangor Darul Ehsan (Malaysia)

2015-08-28

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate [P(3HB-co-3HHx)] produced by C. necator PHB{sup −}4 harboring phaC{sub cs} from crude palm kernel oil with 21 mol% of 3-hydroxyhexanoate and epoxidized natural rubber with 25 mol% of epoxy content (ENR-25) were used to study the miscibility of the blends by attenuated total reflection-Fourier transform infrared (ATR-FTIR) and differential scanning calorimetry (DSC). The polymers used were purified and the blends were prepared by solution casting method. Nuclear magnetic resonance (NMR) spectra confirm the purity and molecular structures of P(3HB-co-3HHx) and ENR-25. FTIR spectra for different compositions of P(3HB-co-3HHx) and ENR-25 blends show absorbance change of the absorbance bands but with no significant shifting of the absorbance bands as the P(3HB-co-3HHx) content decreases, which shows that there is no intermolecular interaction between the parent polymer blends. On top of that, there are two T{sub g}s present for the blends and both remain constant for different compositions which corresponds to the T{sub g}s of the parent polymers. This indicates that the blends are immiscible.

Investigation of miscibility of p(3hydroxybutyrate-co-3hydroxyhexanoate) and epoxidized natural rubber blends

International Nuclear Information System (INIS)

Akram, Faridah; Chan, Chin Han; Natarajan, Valliyappan David

2015-01-01

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate [P(3HB-co-3HHx)] produced by C. necator PHB − 4 harboring phaC cs from crude palm kernel oil with 21 mol% of 3-hydroxyhexanoate and epoxidized natural rubber with 25 mol% of epoxy content (ENR-25) were used to study the miscibility of the blends by attenuated total reflection-Fourier transform infrared (ATR-FTIR) and differential scanning calorimetry (DSC). The polymers used were purified and the blends were prepared by solution casting method. Nuclear magnetic resonance (NMR) spectra confirm the purity and molecular structures of P(3HB-co-3HHx) and ENR-25. FTIR spectra for different compositions of P(3HB-co-3HHx) and ENR-25 blends show absorbance change of the absorbance bands but with no significant shifting of the absorbance bands as the P(3HB-co-3HHx) content decreases, which shows that there is no intermolecular interaction between the parent polymer blends. On top of that, there are two T g s present for the blends and both remain constant for different compositions which corresponds to the T g s of the parent polymers. This indicates that the blends are immiscible

Artificial intelligence based modeling and optimization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production process by using Azohydromonas lata MTCC 2311 from cane molasses supplemented with volatile fatty acids: a genetic algorithm paradigm.

Science.gov (United States)

Zafar, Mohd; Kumar, Shashi; Kumar, Surendra; Dhiman, Amit K

2012-01-01

The present work describes the optimization of medium variables for the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] by Azohydromonas lata MTCC 2311 using cane molasses supplemented with propionic acid. Genetic algorithm (GA) has been used for the optimization of P(3HB-co-3HV) production through the simulation of artificial neural network (ANN) and response surface methodology (RSM). The predictions by ANN are better than those of RSM and in good agreement with experimental findings. The highest P(3HB-co-3HV) concentration and 3HV content have been reported as 7.35 g/l and 16.84 mol%, respectively by hybrid ANN-GA. Upon validation, 7.20 g/l and 16.30 mol% of P(3HB-co-3HV) concentration and 3HV content have been found in the shake flask, whereas 6.70 g/l and 16.35 mol%, have been observed in a 3 l bioreactor, respectively. The specific growth rate and P(3HB-co-3HV) accumulation rate of 0.29 per h and 0.16 g/lh determined with cane molasses are comparable to those observed on pure substrates. Copyright © 2011 Elsevier Ltd. All rights reserved.

Poly-β-hydroxybutyrate sensitizing effect on the photophysical properties of environment friendly fluorescent films containing europium complex

Energy Technology Data Exchange (ETDEWEB)

Yang, Chaolong, E-mail: yclzjun@163.com [School of Materials Science and Engineering, Chongqing University of Technology, Chongqing 400054 (China); Zhang, Pan; Zhou, Hualin [School of Materials Science and Engineering, Chongqing University of Technology, Chongqing 400054 (China); Xu, Jing [Department of Chemistry, Graduate School of Science, Tohoku University, Aramaki-Azaaoba 6-3, Aoba-ku, Sendai (Japan); Li, Youbing [School of Materials Science and Engineering, Chongqing University of Technology, Chongqing 400054 (China); Lu, Mangeng [Key Laboratory of Polymer Materials for Electronics, Guangzhou Institute of Chemistry, Chinese Academy of Sciences, Guangzhou 510650 (China); Lei, Lei; Zhang, Qiang; Zhang, Yi; Chen, Shaopeng [School of Materials Science and Engineering, Chongqing University of Technology, Chongqing 400054 (China)

2016-10-15

A series of environment friendly Eu/PHB fluorescent films through doped the Eu-complex precursor Eu(TTA){sub 2}(Tpy-OCH{sub 3})(2H{sub 2}O) into polymer matrices poly-β-hydroxybutyrate (PHB) with doping percentage at 1, 3, 5, and 7 (mass) were designed, fabricated and characterized. TGA and PL results indicated the Eu-complex precursor was immobilized in PHB matrix through the interaction between the Eu-complex. DSC results indicated the crystallinity of Eu/PHB films decreased with the increase of Eu-complex doping percentage. The emission spectra of the Eu-complex and Eu/PHB films recorded at room temperature exhibited the characteristic bands arising from the {sup 5}D{sub 0}/{sup 7}F{sub J}. The fact that the quantum efficiencies (η) of the doped film increased significantly revealed that the PHB matrix acts as an efficient co-sensitizer for Eu{sup 3+} ions luminescent center and therefore enhances the quantum efficiency of the emitter {sup 5}D{sub 0} level. In particular, all Eu/PHB films can be excited by visible light (410 nm), and also showed good photoluminescent properties. So the new Eu/PHB fluorescent films showed considerable promise for polymer light-emitting diode, active polymer optical fiber and biomedical analysis applications.

Poly-β-hydroxybutyrate sensitizing effect on the photophysical properties of environment friendly fluorescent films containing europium complex

International Nuclear Information System (INIS)

Yang, Chaolong; Zhang, Pan; Zhou, Hualin; Xu, Jing; Li, Youbing; Lu, Mangeng; Lei, Lei; Zhang, Qiang; Zhang, Yi; Chen, Shaopeng

2016-01-01

A series of environment friendly Eu/PHB fluorescent films through doped the Eu-complex precursor Eu(TTA) 2 (Tpy-OCH 3 )(2H 2 O) into polymer matrices poly-β-hydroxybutyrate (PHB) with doping percentage at 1, 3, 5, and 7 (mass) were designed, fabricated and characterized. TGA and PL results indicated the Eu-complex precursor was immobilized in PHB matrix through the interaction between the Eu-complex. DSC results indicated the crystallinity of Eu/PHB films decreased with the increase of Eu-complex doping percentage. The emission spectra of the Eu-complex and Eu/PHB films recorded at room temperature exhibited the characteristic bands arising from the 5 D 0 / 7 F J . The fact that the quantum efficiencies (η) of the doped film increased significantly revealed that the PHB matrix acts as an efficient co-sensitizer for Eu 3+ ions luminescent center and therefore enhances the quantum efficiency of the emitter 5 D 0 level. In particular, all Eu/PHB films can be excited by visible light (410 nm), and also showed good photoluminescent properties. So the new Eu/PHB fluorescent films showed considerable promise for polymer light-emitting diode, active polymer optical fiber and biomedical analysis applications.

Novel poly(3-hydroxybutyrate) nanocomposites containing WS{sub 2} inorganic nanotubes with improved thermal, mechanical and tribological properties

Energy Technology Data Exchange (ETDEWEB)

Naffakh, Mohammed, E-mail: mohammed.naffakh@upm.es [Universidad Politécnica de Madrid, Departamento de Ingeniería y Ciencia de los Materiales, Escuela Técnica Superior de Ingenieros Industriales, José Gutiérrez Abascal 2, 28006 Madrid (Spain); Marco, Carlos; Ellis, Gary [CSIC, Instituto de Ciencia y Tecnología de Polímeros (ICTP), Juan de la Cierva 3, 28006 Madrid (Spain); Cohen, Sidney R. [Department of Chemical Research Support, Weizmann Institute of Science, Rehovot 76100 (Israel); Laikhtman, Alexander; Rapoport, Lev; Zak, Alla [Department of Sciences, Holon Institute of Technology, 52 Golomb St., Holon 58102 (Israel)

2014-09-15

Poly(3-hydroxybutyrate) (PHB) nanocomposites containing environmentally-friendly tungsten disulphide inorganic nanotubes (INT–WS{sub 2}) have been successfully prepared by a simple solution blending method. The dynamic and isothermal crystallization studies by differential scanning calorimetry (DSC) demonstrated that the INT–WS{sub 2} exhibits much more prominent nucleation activity on the crystallization of PHB than specific nucleating agents or other nanoscale fillers. Both crystallization rate and crystallinity significantly increase in the nanocomposites compared to neat PHB. These changes occur without modifying the crystalline structure of PHB in the nanocomposites, as shown by wide-angle X-ray diffraction (WAXS) and infrared/Raman spectroscopy. Other parameters such as the Avrami exponent, the equilibrium melting temperature, global rate constant and the fold surface free energy of PHB chains in the nanocomposites were obtained from the calorimetric data in order to determine the influence of the INT–WS{sub 2} filler. The addition of INT–WS{sub 2} remarkably influences the energetics and kinetics of nucleation and growth of PHB, reducing the fold surface free energy by up to 20%. Furthermore, these nanocomposites also show an improvement in both tribological and mechanical (hardness and modulus) properties with respect to pure PHB evidenced by friction and nanoindentation tests, which is of important potential interest for industrial and medical applications. - Highlights: • Environmentally-friendly INT–WS{sub 2} is used to produce advanced PHB NCPs. • Novel PHB NCPs are obtained without using modifiers or surfactants. • Novel INT–WS{sub 2} improve the thermal and mechanical properties of PHB. • INT–WS{sub 2} is effective in reducing the friction coefficient of PHB. • The benefits of using INTs compared to other nanoscale fillers are highlighted.

Coupled effects of methane monooxygenase and nitrogen source on growth and poly-β-hydroxybutyrate (PHB) production of Methylosinus trichosporium OB3b.

Science.gov (United States)

Zhang, Tingting; Zhou, Jiti; Wang, Xiaowei; Zhang, Yu

2017-02-01

The coupled effects of nitrogen source and methane monooxygenase (MMO) on the growth and poly-β-hydroxybutyrate (PHB) accumulation capacity of methanotrophs were explored. The ammonia-supplied methanotrophs expressing soluble MMO (sMMO) grew at the highest rate, while N 2 -fixing bacteria expressing particulate MMO (pMMO) grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing pMMO, which might cause their slightly lower growth rate. The highest PHB content (51.0%) was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing pMMO. Ammonia-supplied bacteria also accumulated a higher content of PHB (45.2%) with the expression of pMMO, while N 2 -fixing bacteria containing pMMO only showed low PHB production capacity (32.1%). The maximal PHB contents of bacteria expressing sMMO were low, with no significant change under different nitrogen source conditions. The low MMO activity, low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N 2 with the expression of pMMO were greatly improved in the cyclic NO 3 - N 2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing pMMO. Copyright © 2016. Published by Elsevier B.V.

Bioreducible poly(amidoamine)s as carriers for intracellular protein delivery to intestinal cells

NARCIS (Netherlands)

Cohen, S.; Coué, G.M.J.P.C.; Beno, D.; Korenstein, R.; Engbersen, Johannes F.J.

2012-01-01

An effective intracellular protein delivery system was developed based on linear poly(amidoamine)s (PAAs) that form self-assembled cationic nanocomplexes with oppositely charged proteins. Two differently functionalized PAAs were synthesized by Michael-type polyaddition of 4-amino-1-butanol (ABOL) to

Biocomposite scaffolds for bone regeneration: Role of chitosan and hydroxyapatite within poly-3-hydroxybutyrate-co-3-hydroxyvalerate on mechanical properties and in vitro evaluation.

Science.gov (United States)

Zhang, Sai; Prabhakaran, Molamma P; Qin, Xiaohong; Ramakrishna, Seeram

2015-11-01

Bio-engineered scaffolds for bone tissue regeneration is an exploding area of research mainly because they can satisfy the essential demands and current challenges in bone replacement therapies, by imitating the extracellular matrix (ECM) of the native bone. We fabricated bio-composite nanofibrous scaffolds with a blend of poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), chitosan (CTS) and hydroxyapatite (HA) during this study. Morphological evaluation confirmed the fiber diameters of PHBV, PHBV/CTS (90:10), PHBV/CTS/HA4 (85.5:9.5:5) and PHBV/CTS/HA8 (81:9:10) as 405 ± 74 nm, 334 ± 82 nm, 316 ± 103 nm and 256 ± 110 nm, respectively. The PHBV/CTS/HA4 and PHBV/CTS/HA8 scaffolds were capable of enduring the long term culture of human fetal osteoblasts (hFOB) with ultimate tensile strength of 3.55 ± 0.22 MPa and 4.19 ± 0.19 MPa, respectively. The proliferation of osteoblasts on PHBV/CTS/HA8 scaffold was found 34.10% higher than that on PHBV scaffold on day 20. Cell maturation identified by alkaline phosphatase activity on day 20 was significantly higher on PHBV/CTS/HA8 scaffold than that on PHBV scaffold. The cells on PHBV/CTS/HA8 scaffold also acquired higher mineral deposition (25.79%) than the mineral deposition on PHBV scaffold by day 20, confirmed by EDX analysis. Based on the results, we concluded that the electrospun PHBV/CTS/HA8 scaffolds hold great potential to promote the regeneration of bone tissue due to the synergistic effect of chitosan and HA, whereby chitosan provided cell recognition sites while HA acted as a chelating agent for organizing the apatite-like mineralization. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enrichments of methanotrophic-heterotrophic cultures with high poly-β-hydroxybutyrate (PHB) accumulation capacities.

Science.gov (United States)

Zhang, Tingting; Wang, Xiaowei; Zhou, Jiti; Zhang, Yu

2018-03-01

Methanotrophic-heterotrophic communities were selectively enriched from sewage sludge to obtain a mixed culture with high levels of poly-β-hydroxybutyrate (PHB) accumulation capacity from methane. Methane was used as the carbon source, N 2 as sole nitrogen source, and oxygen and Cu content were varied. Copper proved essential for PHB synthesis. All cultures enriched with Cu could accumulate high content of PHB (43.2%-45.9%), while only small amounts of PHB were accumulated by cultures enriched without Cu (11.9%-17.5%). Batch assays revealed that communities grown with Cu and a higher O 2 content synthesized more PHB, which had a wider optimal CH 4 :O 2 range and produced a high PHB content (48.7%) even though in the presence of N 2 . In all methanotrophic-heterotrophic communities, both methanotrophic and heterotrophic populations showed the ability to accumulate PHB. Although methane was added as the sole carbon source, heterotrophs dominated with abundances between 77.2% and 85.6%. All methanotrophs detected belonged to type II genera, which formed stable communities with heterotrophs of different PHB production capacities. Copyright © 2017. Published by Elsevier B.V.

Well-Defined Poly(Ortho Ester Amides) for Potential Drug Carriers: Probing the Effect of Extra- and Intracellular Drug Release on Chemotherapeutic Efficacy.

Science.gov (United States)

Yan, Guoqing; Wang, Jun; Qin, Jiejie; Hu, Liefeng; Zhang, Panpan; Wang, Xin; Tang, Rupei

2017-07-01

To compare the chemotherapeutic efficacy determined by extra- and intracellular drug release strategies, poly(ortho ester amide)-based drug carriers (POEAd-C) with well-defined main-chain lengths, are successfully constructed by a facile method. POEAd-C3-doxorubicin (DOX) can be rapidly dissolved to release drug at tumoral extracellular pH (6.5-7.2), while POEAd-C6-DOX can rapidly release drug following gradual swelling at intracellular pH (5.0-6.0). In vitro cytotoxicity shows that POEAd-C3-DOX exhibits more toxic effect on tumor cells than POEAd-C6-DOX at extracellular pH, but POEAd-C6-DOX has stronger tumor penetration and inhibition in vitro and in vivo tumor models. So, POEAd-C6-DOX with the intracellular drug release strategy has stronger overall chemotherapeutic efficacy than POEAd-C3-DOX with extracellular drug release strategy. It is envisioned that these poly(ortho ester amides) can have great potential as drug carriers for efficient chemotherapy with further optimization. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Burkholderia sacchari DSM 17165: A source of compositionally-tunable block-copolymeric short-chain poly(hydroxyalkanoates) from xylose and levulinic acid

Science.gov (United States)

Burkholderia sacchari DSM 17165 was used as a biocatalyst for the production of poly-3-hydroxybutyrate-co-3-hydroxyvalerate block copolymers (Poly-3HB-block-3HV) from xylose and levulinic acid. Among the carbon source mixtures, levulinic acid was preferred and was consumed early in the fermentations...

Modifications induced by swift heavy ions on poly(hydroxybutyrate-hydroxyvalerate) (PHB/HV) and poly(ε-caprolactone) (PCL). Part 2. Radicals characterization

International Nuclear Information System (INIS)

Rouxhet, L.; Mestdagh, M.; Legras, R.

2000-01-01

Modifications induced by different energetic heavy ions ( 40 Ar 9+ , 80 Kr 15+ , 129 Xe 24+ , 208 Pb 53+ and 208 Pb 56+ ) on poly(ε-caprolactone) (PCL) and poly(hydroxybutyrate-hydroxyvalerate) (PHB/HV) have been investigated by electron spin resonance (ESR). Indeed, film irradiation by heavy ions leads to, among other phenomena, the formation of radicals in the ion tracks. Thanks to ESR, it is possible to detect these radicals and to identify them or at least to characterize them by following the evolution of the radical signal as a function of parameters, like temperature, or the kinetic of disappearance of the radical species at ambient temperature in vacuum or ambient atmosphere. This study confirmed the generation of radicals by the irradiation of PHB/HV samples with energetic heavy ions reported in the literature. The study on PCL was not pursued after a few preliminary studies, revealing the presence of an ESR signal in the non-irradiated sample. Electronic stopping power has a major influence on radical decrease at ambient temperature. The ion used for the irradiation did not modify very much the radical signal and the evolution of the radicalar signal intensity with temperature. Different reasoning and experiments revealed that the glass transition temperature is a key temperature above which irreversible recombinations of the most stable radicals take place. A simulation study indicated that the most stable radical produced was probably a tertiary radical formed by the stabilization of the secondary radical resulting from the abstraction of a highly mobile hydrogen adjacent to the carbonyl

Modifications induced by swift heavy ions in poly(hydroxybutyrate-hydroxyvalerate) (PHB/HV) and poly(ε-caprolactone) (PCL) films. Part 1. Thermal behaviour and molecular mass modifications

International Nuclear Information System (INIS)

Rouxhet, L.; Legras, R.

2000-01-01

Modifications induced by different energetic heavy ions in poly(ε-caprolactone) (PCL) and poly(hydroxybutyrate-hydroxyvalerate) (PHB/HV) have been investigated by the differential scanning calorimetry (DSC) and steric exclusion chromatography (SEC). A certain dose of damages, depending mainly on the charge and mass of the ion and on the intensity of irradiation, has to be overcome in order to detect any effect on PHB/HV. Actually, at a given intensity of irradiation, superior to 10 10 ions/cm 2 , the level of damage intensity increased with the increase in charge and mass of the ion. Moreover, according to the SEC results, there seems to be a critical mass and/or charge threshold above which the dominant type of damages changes. As a matter of fact, high-density irradiation with Ar 9+ and Kr 15+ resulted mainly in chain scission whereas cross-linking was dominant when irradiating the polymer with Xe 24+ and Pb 56+ . The irradiation of PCL in the conditions studied did not modify significantly the values of the melting point, the crystallisation temperature and the molecular masses of the system studied. The main effect of the irradiation detected by the DSC is the cross-linking of the polymer chains

Production and Characterization of Thermostable Bioplastic (Poly-beta-Hydroxybutyrate) from Bacillus Cereus NRRL-B-3711

International Nuclear Information System (INIS)

Asad-ur-Rehman, M.; Aslam, A.; Masood, R.; Aftab, M. N.; Ajmal, R.; Haq, I. U.

2016-01-01

The poly-beta-hydroxybutyrate (PHB) is a thermostable and biocompatible polyester produced by several bacteria under unbalanced nutritional conditions. Among Gram-positive bacteria, which are not well exploited for PHB production on industrial scale, Bacillus appears to be a prospective candidate due to its excellent biopolymer yield and less rigorous fermentation conditions. Batch culture fermentation was carried out for PHB production. The Bacillus cereus NRRL-B-3711 was the most efficient producer of PHB out of five Bacillus species. The optimized production was achieved with 2% glucose, 37 degree C, pH value of 8 and ammonium sulfate (2.5 g/L) as a nitrogen source. Carbon to nitrogen ratio of 10 significantly affects the PHB accumulation. The selected specie was able to accumulate PHB up to 56% (14.2) 0.07 g/L) on dry cell weight basis after optimization. This corresponds to a 1.87 folds increase in the production. The optical microscopy showed extremely flat surface of bioplastic thin film indicating the brittle failure of PHB under tensile loading. The FTIR analysis revealed C=O and -CH groups, with thermal properties e.g. Tg; 2 degree C, Tc; 54 degree C, Tm;162 degree C and percent crystallinity of 51.3 by differential scanning calorimeter (DSC), thus confirming the presence of PHB bioplastic and enhancing its industrial applications.The results surpassed those reported in the literature for PHB production. (author)

Modeling and optimization of poly(3hydroxybutyrate-co-3hydroxyvalerate) production from cane molasses by Azohydromonas lata MTCC 2311 in a stirred-tank reactor: effect of agitation and aeration regimes.

Science.gov (United States)

Zafar, Mohd; Kumar, Shashi; Kumar, Surendra; Dhiman, Amit K

2012-07-01

The effects of agitation and aeration rates on copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] production by Azohydromonas lata MTCC 2311 using cane molasses supplemented with propionic acid in a bioreactor were investigated. The experiments were conducted in a three-level factorial design by varying the impeller (150-500 rev min(-1)) and aeration (0.5-1.5 vvm) rates. Further, the data were fitted to mathematical models [quadratic polynomial equation and artificial neural network (ANN)] and process variables were optimized by genetic algorithm-coupled models. ANN and hybrid ANN-GA were found superior for modeling and optimization of process variables, respectively. The maximum copolymer concentration of 7.45 g l(-1) with 21.50 mol% of 3HV was predicted at process variables: agitation speed, 287 rev min(-1); and aeration rate, 0.85 vvm, which upon validation gave 7.20 g l(-1) of P(3HB-co-3HV) with 21 mol% of 3HV with the prediction error (%) of 3.38 and 2.32, respectively. Agitation speed established a relative high importance of 72.19% than of aeration rate (27.80%) for copolymer accumulation. The volumetric gas-liquid mass transfer coefficient (k (L) a) was strongly affected by agitation and aeration rates. The highest P(3HB-co-3HV) productivity of 0.163 g l(-1) h(-1) was achieved at 0.17 s(-1) of k (L) a value. During the early phase of copolymer production process, 3HB monomers were accumulated, which were shifted to 3HV units (9-21%) during the cultivation period of 24-42 h. The enhancement of 7.5 and 34% were reported for P(3HB-co-3HV) production and 3HV content, respectively, by hybrid ANN-GA paradigm, which revealed the significant utilization of cane molasses for improved copolymer production.

Poly (hydroxybutyrate co hydroxyvalerate Nanofibrous Scaffold Containing HydroxyapatiteBredigite Nanoparticles: Characterization and Biological Evaluation

Directory of Open Access Journals (Sweden)

M. Kouhi

2017-11-01

Full Text Available In this work, poly (hydroxybutyrate co hydroxyvalerate (PHBV composite nanofibrous scaffold containing hydroxyapatite/bredigite (HABR nanoparticles was fabricated through electrospining method. The morphology of prepared  nanofibers and the state of the nanoparticles dispersion in nanofiber matrix were investigated using scanning and transmission electron microscopy, respectively. Evaluation of the mechanical properties of the nanofibrous scaffolds revealed that there is a limit to the nanoparticle concentration at which nanoparticles can improve the mechanical properties of the nanofibrous scaffolds. According to the results, PHBV/HABR nanofibers showed higher wettability compared to PHBV nanofibers. In vitro cell culture assay was done using human fetal osteoblast cells on nanofibrous scaffold. MTS assay revealed that cell proliferation on the composite nanofibrous scaffold was significantly higher than those on the pure scaffold after 10 and 15 days. Scanning electron microscopy- Energy dispersive X-ray spectroscopy and CMFDA colorimeter assay analysis showed that the cells on the PHBV/HABR scaffolds acquired higher mineral deposition than the cells on the pure PHBV and control sample scaffold. Based on the results we concluded that PHBV/HABR nanofibers scaffold with higher wettability, improved mechanical properties and cell behavior hold great potential in bone regeneration applications.

Expression of the Azotobacter vinelandii Poly-β-Hydroxybutyrate Biosynthetic phbBAC Operon Is Driven by Two Overlapping Promoters and Is Dependent on the Transcriptional Activator PhbR

OpenAIRE

Peralta-Gil, Martín; Segura, Daniel; Guzmán, Josefina; Servín-González, Luis; Espín, Guadalupe

2002-01-01

The Azotobacter vinelandii phbBAC genes encode the enzymes for poly-β-hydroxybutyrate (PHB) synthesis. The phbR gene, which is located upstream of and in the opposite direction of phbBAC, encodes PhbR, a transcriptional activator which is a member of the AraC family of activators. Here we report that a mutation in phbR reduced PHB accumulation and transcription of a phbB-lacZ fusion. We also report that phbB is transcribed from two overlapping promoters, pB1 and pB2. The region corresponding ...

Global changes in the proteome of Cupriavidus necator H16 during poly-(3-hydroxybutyrate) synthesis from various biodiesel by-product substrates.

Science.gov (United States)

Sharma, Parveen K; Fu, Jilagamazhi; Spicer, Victor; Krokhin, Oleg V; Cicek, Nazim; Sparling, Richard; Levin, David B

2016-12-01

Synthesis of poly-[3-hydroxybutyrate] (PHB) by Cupriavidus necator H16 in batch cultures was evaluated using three biodiesel-derived by-products as the sole carbon sources: waste glycerol (REG-80, refined to 80 % purity with negligible free fatty acids); glycerol bottom (REG-GB, with up to 65 % glycerol and 35 % free fatty acids), and free fatty acids (REG-FFA, with up to 75 % FFA and no glycerol). All the three substrates supported growth and PHB production by C. necator, with polymer accumulation ranging from 9 to 84 % cell dry weight (cdw), depending on the carbon source. To help understand these differences, proteomic analysis indicated that although C. necator H16 was able to accumulate PHB during growth on all three biodiesel by-products, no changes in the levels of PHB synthesis enzymes were observed. However, significant changes in the levels of expression were observed for two Phasin proteins involved with PHB accumulation, and for a number of gene products in the fatty acid β-oxidation pathway, the Glyoxylate Shunt, and the hydrogen (H2) synthesis pathways in C. necator cells cultured with different substrates. The glycerol transport protein (GlpF) was induced in REG-GB and REG-80 glycerol cultures only. Cupriavidus necator cells cultured with REG-GB and REG-FFA showed up-regulation of β-oxidation and Glyoxylate Shunt pathways proteins at 24 h pi, but H2 synthesis pathways enzymes were significantly down-regulated, compared with cells cultured with waste glycerol. Our data confirmed earlier observations of constitutive expression of PHB synthesis proteins, but further suggested that C. necator H16 cells growing on biodiesel-derived glycerol were under oxidative stress.

Nanoparticles of poly(hydroxybutyrate-co-hydroxyvalerate) as support for the immobilization of Candida antarctica lipase (fraction B)

International Nuclear Information System (INIS)

Fernandes, Ilizandra A.; Nyari, Nadia L.D.; Oliveira, Jose Vladimir de; Oliveira, Debora de; Rigo, Elisandra; Souza, Maria Cristiane M. de; Goncalves, Luciana R.B.; Pergher, Sibele Berenice C.

2014-01-01

This work evaluates the immobilization of Candida antarctica lipase (Fraction B) using poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) nanoparticles as support. The effects of immobilization time (30-150 min) and pH (5-10) on lipase loading were evaluated. The stability of the immobilized enzyme towards temperature (40, 60, and 80 deg C), reuse and storage (at 4 deg C) were also determined. Furthermore, to assess its potential application in a system of interest, the immobilized lipase was used as a catalyst in the esterification of geraniol with oleic acid. The results indicated a time of 120 minutes and pH of 7 as optimal for immobilization. A 21 hour exposure of the PHBV-lipase derivative to 60 deg C showed a 33% reduction of the initial activity while storage at 4 deg C led to a residual activity (5% of the original activity). The derivative was used without significant loss of activity for 4 successive cycles. The use of the immobilized lipase as a catalyst in the production of geranyl oleate led to about 88% conversion of the initial reactants to products. (author)

Evaluation of structural and mechanical properties of electrospun nano-micro hybrid of poly hydroxybutyrate-chitosan/silk scaffold for cartilage tissue engineering.

Science.gov (United States)

Karbasi, Saeed; Fekrat, Farnoosh; Semnani, Daryoush; Razavi, Shahnaz; Zargar, Elham Naghash

2016-01-01

One of the new methods of scaffold fabrication is a nano-micro hybrid structure in which the properties of the scaffold are improved by introducing nanometer and micrometer structures. This method could be suitable for scaffold designing if some features improve. In this study, electrospun nanofibers of 9% weight solution of poly (3-hydroxybutyrate) (P3HB) and a 15% weight of chitosan by trifluoroacetic acid were coated on both the surface of a silk knitted substrate in the optimum condition to improve the mechanical properties of scaffolds for cartilage tissue engineering application. These hybrid nano-micro fibrous scaffolds were characterized by structural and mechanical evaluation methods. Scanning electron microscopy values and porosity analysis showed that average diameter of nanofibers was 584.94 nm in electrospinning part and general porosity was more than 80%. Fourier transform infrared spectroscopy results indicated the presence of all elements without pollution. The tensile test also stated that by electrospinning, as well as adding chitosan, both maximum strength and maximum elongation increased to 187 N and 10 mm. It means that the microfibrous part of scaffold could affect mechanical properties of nano part of the hybrid scaffold, significantly. It could be concluded that P3HB-chitosan/silk hybrid scaffolds can be a good candidate for cartilage tissue engineering.

Poly(3-hydroxybutyrate)/caffeic acid electrospun fibrous materials coated with polyelectrolyte complex and their antibacterial activity and in vitro antitumor effect against HeLa cells

Energy Technology Data Exchange (ETDEWEB)

Ignatova, Milena G. [Laboratory of Bioactive Polymers, Institute of Polymers, Bulgarian Academy of Sciences, Acad. G. Bonchev St, Bl. 103A, BG-1113 Sofia (Bulgaria); Manolova, Nevena E., E-mail: manolova@polymer.bas.bg [Laboratory of Bioactive Polymers, Institute of Polymers, Bulgarian Academy of Sciences, Acad. G. Bonchev St, Bl. 103A, BG-1113 Sofia (Bulgaria); Rashkov, Iliya B. [Laboratory of Bioactive Polymers, Institute of Polymers, Bulgarian Academy of Sciences, Acad. G. Bonchev St, Bl. 103A, BG-1113 Sofia (Bulgaria); Markova, Nadya D. [Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev Bl. 26, BG-1113 Sofia (Bulgaria); Toshkova, Reneta A.; Georgieva, Ani K.; Nikolova, Elena B. [Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Acad. G. Bonchev St, bl. 25, BG-1113 Sofia (Bulgaria)

2016-08-01

The purpose of this work was to investigate the possibility for the preparation of new poly(3-hydroxybutyrate) (PHB)/poly(ethylene glycol) (PEG)-based fibrous materials containing natural phenolic compound caffeic acid (CA) of diverse architectures, as well as to study the impact of the fiber composition on the in vitro CA release profile and on the biological properties of the fibrous materials. The application of the one-pot electrospinning enabled the fabrication of nanofibrous materials from PHB and PEG loaded with the CA. Materials with targeted design were obtained by coating with polyelectrolyte complex of alginate (Alg) and N,N,N-trimethylchitosan (TMCh). Three different processing paths were used to obtain coated mats: (i) with CA incorporated in the PHB/PEG core; (ii) with CA embedded in the Alg layer; and (iii) with CA included in the TMCh layer. The in vitro release of CA was modulated by controlling the composition and the architecture of the nanofibrous mats. The performed microbiological screening and MTT cell viability studies revealed that in contrast to the bare mats, the CA-containing nanofibrous materials were effective in suppressing the growth of the Gram-positive bacteria Staphylococcus aureus and the Gram-negative bacteria Escherichia coli and displayed good cytotoxicity against human cervical HeLa tumor cells. In addition, the proliferation of murine spleen lymphocytes and peritoneal macrophages was increased by the prepared CA-containing nanofibrous materials. The obtained materials are promising for antibacterial wound dressing applications as well as for application in local treatment of cervical tumors. - Highlights: • New caffeic acid-loaded materials from PHB and PEG were prepared by electrospinning. • Different design is achieved by coating and formation of polyelectrolyte complexes. • The control on the architecture of the mats enables modulating caffeic acid release. • The caffeic acid-loaded mats suppress the growth of

Poly(3-hydroxybutyrate)/caffeic acid electrospun fibrous materials coated with polyelectrolyte complex and their antibacterial activity and in vitro antitumor effect against HeLa cells

International Nuclear Information System (INIS)

Ignatova, Milena G.; Manolova, Nevena E.; Rashkov, Iliya B.; Markova, Nadya D.; Toshkova, Reneta A.; Georgieva, Ani K.; Nikolova, Elena B.

2016-01-01

The purpose of this work was to investigate the possibility for the preparation of new poly(3-hydroxybutyrate) (PHB)/poly(ethylene glycol) (PEG)-based fibrous materials containing natural phenolic compound caffeic acid (CA) of diverse architectures, as well as to study the impact of the fiber composition on the in vitro CA release profile and on the biological properties of the fibrous materials. The application of the one-pot electrospinning enabled the fabrication of nanofibrous materials from PHB and PEG loaded with the CA. Materials with targeted design were obtained by coating with polyelectrolyte complex of alginate (Alg) and N,N,N-trimethylchitosan (TMCh). Three different processing paths were used to obtain coated mats: (i) with CA incorporated in the PHB/PEG core; (ii) with CA embedded in the Alg layer; and (iii) with CA included in the TMCh layer. The in vitro release of CA was modulated by controlling the composition and the architecture of the nanofibrous mats. The performed microbiological screening and MTT cell viability studies revealed that in contrast to the bare mats, the CA-containing nanofibrous materials were effective in suppressing the growth of the Gram-positive bacteria Staphylococcus aureus and the Gram-negative bacteria Escherichia coli and displayed good cytotoxicity against human cervical HeLa tumor cells. In addition, the proliferation of murine spleen lymphocytes and peritoneal macrophages was increased by the prepared CA-containing nanofibrous materials. The obtained materials are promising for antibacterial wound dressing applications as well as for application in local treatment of cervical tumors. - Highlights: • New caffeic acid-loaded materials from PHB and PEG were prepared by electrospinning. • Different design is achieved by coating and formation of polyelectrolyte complexes. • The control on the architecture of the mats enables modulating caffeic acid release. • The caffeic acid-loaded mats suppress the growth of

Poly(hydroxybutyrate)/cellulose acetate blend nanofiber scaffolds: Preparation, characterization and cytocompatibility

Energy Technology Data Exchange (ETDEWEB)

Zhijiang, Cai, E-mail: caizhijiang@hotmail.com [School of Textiles, Tianjin Polytechnic University, Tianjin 300387 (China); State Key Laboratory of Hollow Fiber Membrane Material and Processes, No 399 BingShuiXi Street, XiQing District, Tianjin, China, 300387 (China); Yi, Xu; Haizheng, Yang; Jia, Jianru; Liu, Yuanpei [School of Textiles, Tianjin Polytechnic University, Tianjin 300387 (China)

2016-01-01

Poly(hydroxybutyrate) (PHB)/cellulose acetate (CA) blend nanofiber scaffolds were fabricated by electrospinning using the blends of chloroform and DMF as solvent. The blend nanofiber scaffolds were characterized by SEM, FTIR, XRD, DSC, contact angle and tensile test. The blend nanofibers exhibited cylindrical, uniform, bead-free and random orientation with the diameter ranged from 80–680 nm. The scaffolds had very well interconnected porous fibrous network structure and large aspect surface areas. It was found that the presence of CA affected the crystallization of PHB due to formation of intermolecular hydrogen bonds, which restricted the preferential orientation of PHB molecules. The DSC result showed that the PHB and CA were miscible in the blend nanofiber. An increase in the glass transition temperature was observed with increasing CA content. Additionally, the mechanical properties of blend nanofiber scaffolds were largely influenced by the weight ratio of PHB/CA. The tensile strength, yield strength and elongation at break of the blend nanofiber scaffolds increased from 3.3 ± 0.35 MPa, 2.8 ± 0.26 MPa, and 8 ± 0.77% to 5.05 ± 0.52 MPa, 4.6 ± 0.82 MPa, and 17.6 ± 1.24% by increasing PHB content from 60% to 90%, respectively. The water contact angle of blend nanofiber scaffolds decreased about 50% from 112 ± 2.1° to 60 ± 0.75°. The biodegradability was evaluated by in vitro degradation test and the results revealed that the blend nanofiber scaffolds showed much higher degradation rates than the neat PHB. The cytocompatibility of the blend nanofiber scaffolds was preliminarily evaluated by cell adhesion studies. The cells incubated with PHB/CA blend nanofiber scaffold for 48 h were capable of forming cell adhesion and proliferation. It showed much better biocompatibility than pure PHB film. Thus, the prepared PHB/CA blend nanofiber scaffolds are bioactive and may be more suitable for cell proliferation suggesting that these scaffolds can be used for

Biodegradability of Poly(hydroxyalkanoate Materials

Directory of Open Access Journals (Sweden)

Keiji Numata

2009-08-01

Full Text Available Poly(hydroxyalkanoate (PHA, which is produced from renewable carbon resources by many microorganisms, is an environmentally compatible polymeric material and can be processed into films and fibers. Biodegradation of PHA material occurs due to the action of extracellular PHA depolymerase secreted from microorganisms in various natural environments. A key step in determining the overall enzymatic or environmental degradation rate of PHA material is the degradation of PHA lamellar crystals in materials; hence, the degradation mechanism of PHA lamellar crystals has been studied in detail over the last two decades. In this review, the relationship between crystal structure and enzymatic degradation behavior, in particular degradation rates, of films and fibers for PHA is described.

Production of poly-beta-hydroxybutyric acid by microorganisms accumulated from river water using a two-stage perfusion culture system.

Science.gov (United States)

Morimoto, T; Tashiro, F; Nagashima, H; Nishizawa, K; Nagata, F; Yokogawa, Y; Suzuki, T

2000-01-01

The perfusion culture system using a shaken ceramic membrane flask (SCMF) was employed to accumulate microorganisms separated from river water and to produce poly-beta-hydroxybutyric acid (PHB). Using a two-step culture method with a single SCMF, river microorganisms were cultured by separately feeding four representative carbon sources, n-propanol, lactic acid, methanol, and formic acid. After 140 h culture, the cell concentration and PHB content respectively reached 43 g/l and 35% when a propanol medium was fed. Using a two-stage perfusion culture with twin SCMFs, the seed cell mass was increased in the first SCMF and then supplied to the second flask for PHB production. As a consequence, the cellular PHB content rose to 51% in the second SCMF, while the cell concentration gradually increased to 25 g/l after 175 h perfusion culture. These results demonstrated the utility of the two-stage perfusion culture system for developing a cheap means of producing PHB coincident with wastewater treatment.

Randomized, Multicenter, Double-Blind Study of the Safety and Efficacy of 1%D-3-Hydroxybutyrate eye drops for Dry Eye Disease.

Science.gov (United States)

Kawakita, Tetsuya; Uchino, Miki; Fukagawa, Kazumi; Yoshino, Kenichi; Shimazaki, Seika; Toda, Ikuko; Tanaka, Mari; Arai, Hiroyuki; Sakatani, Keiko; Hata, Seiichiro; Okano, Takashi; Tsubota, Kazuo

2016-02-11

In a previous study, we demonstrated that topical D-beta-hydroxybutyrate ameliorates corneal epithelial erosion and superficial punctate keratopathy in a rat model of dry eye disease. In the current investigation, we performed a prospective, randomized, multicentre, double-blind, placebo-controlled study to assess the safety and efficacy of 1% D-3-hydroxybutyrate eye drops in patients with dry eye disease. A total of 65 patients were randomly assigned to either the placebo group or the 1% D-3-hydroxybutyrate group, and the treatments were administered 6 times a day for 4 weeks. We then evaluated corneal fluorescein staining, corneal and conjunctival rose Bengal staining, tear film break-up time (BUT), Schirmer score, and subjective symptoms. At both 2 and 4 weeks, the corneal rose Bengal score was significantly better in the 1% D-3-hydroxybutyrate group than in the placebo group. Among patients with an initial Schirmer score of ≤5 mm, the corneal fluorescein staining score was significantly better in the 1% D-3-hydroxybutyrate group than in the placebo group at two weeks. Mild ocular symptoms occurred in both groups, and these spontaneously resolved. The present study suggested that 1% D-3-hydroxybutyrate eye drops are safe and effective in treating ocular surface disorders in patients with tear-deficient dry eye disease.

Wastewater Utilization for Poly-β-Hydroxybutyrate Production by the Cyanobacterium Aulosira fertilissima in a Recirculatory Aquaculture System▿

Science.gov (United States)

Samantaray, Shilalipi; Nayak, Jitendra Kumar; Mallick, Nirupama

2011-01-01

Intensive aquaculture releases large quantities of nutrients into aquatic bodies, which can lead to eutrophication. The objective of this study was the development of a biological recirculatory wastewater treatment system with a diazotrophic cyanobacterium, Aulosira fertilissima, and simultaneous production of valuable product in the form of poly-β-hydroxybutyrate (PHB). To investigate this possible synergy, batch scale tests were conducted under a recirculatory aquaculture system in fiber-reinforced plastic tanks enhanced by several manageable parameters (e.g., sedimentation, inoculum size, depth, turbulence, and light intensity), an adequate combination of which showed better productivity. The dissolved-oxygen level increased in the range of 3.2 to 6.9 mg liter−1 during the culture period. Nutrients such as ammonia, nitrite, and phosphate decreased to as low as zero within 15 days of incubation, indicating the system's bioremediation capability while yielding valuable cyanobacterial biomass for PHB production. Maximum PHB accumulation in A. fertilissima was found in sedimented fish pond discharge at 20-cm culture depth with stirring and an initial inoculum size of 80 mg dry cell weight (dcw) liter−1. Under optimized conditions, the PHB yield was boosted to 92, 89, and 80 g m−2, respectively for the summer, rainy, and winter seasons. Extrapolation of the result showed that a hectare of A. fertilissima cultivation in fish pond discharge would give an annual harvest of ∼17 tons dry biomass, consisting of 14 tons of PHB with material properties comparable to those of the bacterial polymer, with simultaneous treatment of 32,640 m3 water discharge. PMID:21984242

Integration of poly-3-(hydroxybutyrate-co-hydroxyvalerate) production by Haloferax mediterranei through utilization of stillage from rice-based ethanol manufacture in India and its techno-economic analysis.

Science.gov (United States)

Bhattacharyya, Anirban; Jana, Kuntal; Haldar, Saubhik; Bhowmic, Asit; Mukhopadhyay, Ujjal Kumar; De, Sudipta; Mukherjee, Joydeep

2015-05-01

Haloferax mediterranei has potential for economical industrial-scale production of polyhydroxyalkanoate (PHA) as it can utilize cheap carbon sources, has capacity for nonsterile cultivation and allows simple product recovery. Molasses-based Indian distilleries are converting themselves to cereal-based distilleries. Waste stillage (14 l) of rice-based ethanol industry was used for the production of PHA by H. mediterranei in the simple plug-flow reactor configuration of the activated sludge process. Cells utilized stillage and accumulated 63 ± 3 % PHA of dry cell weight and produced 13.12 ± 0.05 g PHA/l. The product yield coefficient was 0.27 while 0.14 g/l h volumetric productivity was reached. Simultaneous lowering of 5-day biochemical oxygen demand and chemical oxygen demand values of stillage by 82 % was attained. The biopolymer was characterized as poly-3-(hydroxybutyrate-co-17.9 mol%-hydroxyvalerate) (PHBV). Directional properties of decanoic acid jointly with temperature-dependent water solubility in decanoic acid were employed for two-step desalination of the spent stillage medium in a cylindrical baffled-tank with an immersed heater and a stirrer holding axial and radial impellers. 99.3 % of the medium salts were recovered and re-used for PHA production. The cost of PHBV was estimated as US$2.05/kg when the annual production was simulated as 1890 tons. Desalination contributed maximally to the overall cost. Technology and cost-analysis demonstrate that PHA production integrated with ethanol manufacture is feasible in India. This study could be the basis for construction of a pilot plant.

Production of (R)-3-hydroxybutyric acid by Arxula adeninivorans.

Science.gov (United States)

Biernacki, Mateusz; Riechen, Jan; Hähnel, Urs; Roick, Thomas; Baronian, Kim; Bode, Rüdiger; Kunze, Gotthard

2017-12-01

(R)-3-hydroxybutyric acid can be used in industrial and health applications. The synthesis pathway comprises two enzymes, β-ketothiolase and acetoacetyl-CoA reductase which convert cytoplasmic acetyl-CoA to (R)-3-hydroxybutyric acid [(R)-3-HB] which is released into the culture medium. In the present study we used the non-conventional yeast, Arxula adeninivorans, for the synthesis enantiopure (R)-3-HB. To establish optimal production, we investigated three different endogenous yeast thiolases (Akat1p, Akat2p, Akat4p) and three bacterial thiolases (atoBp, thlp, phaAp) in combination with an enantiospecific reductase (phaBp) from Cupriavidus necator H16 and endogenous yeast reductases (Atpk2p, Afox2p). We found that Arxula is able to release (R)-3-HB used an existing secretion system negating the need to engineer membrane transport. Overexpression of thl and phaB genes in organisms cultured in a shaking flask resulted in 4.84 g L -1 (R)-3-HB, at a rate of 0.023 g L -1  h -1 over 214 h. Fed-batch culturing with glucose as a carbon source did not improve the yield, but a similar level was reached with a shorter incubation period [3.78 g L -1 of (R)-3-HB at 89 h] and the rate of production was doubled to 0.043 g L -1  h -1 which is higher than any levels in yeast reported to date. The secreted (R)-3-HB was 99.9% pure. This is the first evidence of enantiopure (R)-3-HB synthesis using yeast as a production host and glucose as a carbon source.

An Electrochemical Enzyme Biosensor for 3-Hydroxybutyrate Detection Using Screen-Printed Electrodes Modified by Reduced Graphene Oxide and Thionine

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Gonzalo Martínez-García

2017-11-01

Full Text Available A biosensor for 3-hydroxybutyrate (3-HB involving immobilization of the enzyme 3-hydroxybutyrate dehydrogenase onto a screen-printed carbon electrode modified with reduced graphene oxide (GO and thionine (THI is reported here. After addition of 3-hydroxybutyrate or the sample in the presence of NAD+ cofactor, the generated NADH could be detected amperometrically at 0.0 V vs. Ag pseudo reference electrode. Under the optimized experimental conditions, a calibration plot for 3-HB was constructed showing a wide linear range between 0.010 and 0.400 mM 3-HB which covers the clinically relevant levels for diluted serum samples. In addition, a limit of detection of 1.0 µM, much lower than that reported using other biosensors, was achieved. The analytical usefulness of the developed biosensor was demonstrated via application to spiked serum samples.

Improved productivity of poly (3-hydroxybutyrate) (PHB) in thermophilic Chelatococcus daeguensis TAD1 using glycerol as the growth substrate in a fed-batch culture.

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Cui, Bin; Huang, Shaobin; Xu, Fuqian; Zhang, Ruijian; Zhang, Yongqing

2015-07-01

A particularly successful polyhydroxyalkanoate (PHA) in industrial applications is poly (3-hydroxybutyrate) (PHB). However, one of the major obstacles for wider application of PHB is the cost of its production and purification. Therefore, it is desirable to discover a method for producing PHB in large quantities at a competitive price. Glycerol is a cheap and widely used carbon source that can be applied in PHB production process. There are numerous advantages to operating fermentation at elevated temperatures; only several thermophilic bacteria are able to accumulate PHB when glycerol is the growth substrate. Here, we report on the possibility of increasing PHB production at low cost using thermophilic Chelatococcus daeguensis TAD1 when glycerol is the growth substrate in a fed-batch culture. We found that (1) excess glycerol inhibited PHB accumulation and (2) organic nitrogen sources, such as tryptone and yeast extract, promoted the growth of C. daeguensis TAD1. In the batch fermentation experiments, we found that using glycerol at low concentrations as the sole carbon source, along with the addition of mixed nitrate (NH4Cl, tryptone, and yeast extract), stimulated PHB accumulation in C. daeguensis TAD1. The results showed that the PHB productivity decreased in the following order: two-stage fed-batch fermentation > fed-batch fermentation > batch fermentation. In optimized culture conditions, a PHB amount of 17.4 g l(-1) was obtained using a two-stage feeding regimen, leading to a productivity rate of 0.434 g l(-1) h(-1), which is the highest productivity rate reported for PHB to date. This high PHB biosynthetic productivity could decrease the total production cost, allowing for further development of industrial applications of PHB.

Production of poly-3-(hydroxybutyrate-co-hydroxyvalerate) by Haloferax mediterranei using rice-based ethanol stillage with simultaneous recovery and re-use of medium salts.

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Bhattacharyya, Anirban; Saha, Jayeeta; Haldar, Saubhik; Bhowmic, Asit; Mukhopadhyay, Ujjal Kumar; Mukherjee, Joydeep

2014-03-01

Haloferax mediterranei holds promise for competitive industrial-scale production of polyhydroxyalkanoate (PHA) because cheap carbon sources can be used thus lowering production costs. Although high salt concentration in production medium permits a non-sterile, low-cost process, salt disposal after process completion is a problem as current environmental standards do not allow total dissolved solids (TDS) above 2000 mg/l in discharge water. As the first objective of this work, the waste product of rice-based ethanol industry, stillage, was used for the production of PHA by H. mediterranei in shake flasks. Utilization of raw stillage led to 71 ± 2% (of dry cell weight) PHA accumulation and 16.42 ± 0.02 g/l PHA production. The product yield coefficient was 0.35 while 0.17 g/l h volumetric productivity was attained. Simultaneous reduction of BOD5 and COD values of stillage by 83% was accomplished. The PHA was isolated by osmotic lysis of cells, purification by sodium dodecyl sulfate and organic solvents. The biopolymer was identified as poly-3-(hydroxybutyrate-co-15.4 mol%-hydroxyvalerate) (PHBV). This first report on utilization of rice-based ethanol stillage for PHBV production by H. mediterranei is currently the most cost effective. As the second objective, directional properties of decanoic acid together with temperature dependence of water solubility in decanoic acid were applied for two-stage desalination of the spent stillage medium. We report for the first time, recovery and re-use of 96% of the medium salts for PHA production thus removing the major bottleneck in the potential application of H. mediterranei for industrial production of PHBV. Final discharge water had TDS content of 670 mg/l.

Direct Production of Propene from the Thermolysis of Poly(β-hydroxybutyrate) (PHB). An Experimental and DFT Investigation.

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Clark, Jared M; Pilath, Heidi M; Mittal, Ashutosh; Michener, William E; Robichaud, David J; Johnson, David K

2016-01-28

We demonstrate a synthetic route toward the production of propene directly from poly(β-hydroxybutyrate) (PHB), the most common of a wide range of high-molecular-mass microbial polyhydroxyalkanoates. Propene, a major commercial hydrocarbon, was obtained from the depolymerization of PHB and subsequent decarboxylation of the crotonic acid monomer in good yields (up to 75 mol %). The energetics of PHB depolymerization and the gas-phase decarboxylation of crotonic acid were also studied using density functional theory (DFT). The average activation energy for the cleavage of the R'C(O)O-R linkage is calculated to be 163.9 ± 7.0 kJ mol(-1). Intramolecular, autoacceleration effects regarding the depolymerization of PHB, as suggested in some literature accounts, arising from the formation of crotonyl and carboxyl functional groups in the products could not be confirmed by the results of DFT and microkinetic modeling. DFT results, however, suggest that intermolecular catalysis involving terminal carboxyl groups may accelerate PHB depolymerization. Activation energies for this process were estimated to be about 20 kJ mol(-1) lower than that for the noncatalyzed ester cleavage, 144.3 ± 6.4 kJ mol(-1). DFT calculations predict the decarboxylation of crotonic acid to follow second-order kinetics with an activation energy of 147.5 ± 6.3 kJ mol(-1), consistent with that measured experimentally, 146.9 kJ mol(-1). Microkinetic modeling of the PHB to propene overall reaction predicts decarboxylation of crotonic acid to be the rate-limiting step, consistent with experimental observations. The results also indicate that improvements made to enhance the isomerization of crotonic acid to vinylacetic acid will improve the direct conversion of PHB to propene.

Processing recommendations for using low-solids digestate as nutrient solution for poly-ß-hydroxybutyrate production with Synechocystis salina.

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Meixner, K; Fritz, I; Daffert, C; Markl, K; Fuchs, W; Drosg, B

2016-12-20

Within the last decades, environmental pollution with persistent plastics steadily increased; therefore the production of biodegradable materials like poly-ß-hydroxybutyrate (PHB) is essential. Currently, PHB is produced with heterotrophic bacteria from crops. This leads to competition with food and feed production, which can be avoided by using photoautotrophic cyanobacteria, as Synechocystis salina, synthesizing PHB from CO 2 at nutrient limitation. This study aims to increase the economic efficiency of PHB production with cyanobacteria by using nutrients from anaerobic digestate. First, growth and PHB production of S. salina in digestate fractions (supernatant and permeate, with/without precipitating agents) and dilutions thereof and then the scale-up (photobioreactor, 200 L working volume) were evaluated. With precipitated and centrifuged digestate diluted 1/3 the highest biomass (1.55gL -1 ) and PHB concentrations (95.4mgL -1 ), being 78% of those in mineral media, were achieved. In the photobioreactor-experiments biomass (1.63gL -1 ) and PHB concentrations (88.7mgL -1 ), being 79% and 72% of those in mineral medium, were reached, but in a cultivation time 10days longer than in mineral medium. The possibility to use digestate as sustainable and low cost nutrient solution for microalgae cultivation and photoautotrophic PHB production, instead of applying it on fields or processing it to achieve discharge limits, makes this application a highly valid option. Copyright © 2016 Elsevier B.V. All rights reserved.

Wastewater utilization for poly-β-hydroxybutyrate production by the cyanobacterium Aulosira fertilissima in a recirculatory aquaculture system.

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Samantaray, Shilalipi; Nayak, Jitendra Kumar; Mallick, Nirupama

2011-12-01

Intensive aquaculture releases large quantities of nutrients into aquatic bodies, which can lead to eutrophication. The objective of this study was the development of a biological recirculatory wastewater treatment system with a diazotrophic cyanobacterium, Aulosira fertilissima, and simultaneous production of valuable product in the form of poly-β-hydroxybutyrate (PHB). To investigate this possible synergy, batch scale tests were conducted under a recirculatory aquaculture system in fiber-reinforced plastic tanks enhanced by several manageable parameters (e.g., sedimentation, inoculum size, depth, turbulence, and light intensity), an adequate combination of which showed better productivity. The dissolved-oxygen level increased in the range of 3.2 to 6.9 mg liter⁻¹ during the culture period. Nutrients such as ammonia, nitrite, and phosphate decreased to as low as zero within 15 days of incubation, indicating the system's bioremediation capability while yielding valuable cyanobacterial biomass for PHB production. Maximum PHB accumulation in A. fertilissima was found in sedimented fish pond discharge at 20-cm culture depth with stirring and an initial inoculum size of 80 mg dry cell weight (dcw) liter⁻¹. Under optimized conditions, the PHB yield was boosted to 92, 89, and 80 g m⁻², respectively for the summer, rainy, and winter seasons. Extrapolation of the result showed that a hectare of A. fertilissima cultivation in fish pond discharge would give an annual harvest of ∼17 tons dry biomass, consisting of 14 tons of PHB with material properties comparable to those of the bacterial polymer, with simultaneous treatment of 32,640 m³ water discharge.

3-Hydroxybutyrate regulates energy metabolism and induces BDNF expression in cerebral cortical neurons

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Marosi, Krisztina; Kim, Sang Woo; Moehl, Keelin

2016-01-01

During fasting and vigorous exercise, a shift of brain cell energy substrate utilization from glucose to the ketone 3-hydroxybutyrate (3OHB) occurs. Studies have shown that 3OHB can protect neurons against excitotoxicity and oxidative stress, but the underlying mechanisms remain unclear. Neurons ...... suggest cellular signaling mechanisms by which 3OHB may mediate adaptive responses of neurons to fasting, exercise, and ketogenic diets....

Biosynthesis and composition of bacterial poly(hydroxyalkanoates).

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Anderson, A J; Haywood, G W; Dawes, E A

1990-04-01

It is well established that Alcaligenes eutrophus can accumulate a copolymer containing 3-hydroxybutyrate and 3-hydroxyvalerate, but longer 3-hydroxyacid monomers have not been reported to occur in this organism. The properties of the enzymes of poly(hydroxyalkanoate) (PHA) biosynthesis are discussed and it is proposed that the substrate specificity of the polymerizing enzyme restricts the range of monomer units incorporated into PHA. Various other bacteria produce similar copolymers from propionic acid and/or valeric acid. A number of Pseudomonas species accumulate PHAs containing longer-chain monomer units from linear alkanoic acids, alkanes and alcohols.

COST-EFFECTIVE PRODUCTION OF THE BIO-PLASTIC POLY-β-HYDROXYBUTYRATE USING ACINETOBACTER BAUMANNII ISOLATE P39

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Noha Salah Elsayed

2016-06-01

Full Text Available Being biodegradable and biocompatible natural polymer, poly-β-hydroxybutyrate (PHB drew the attention of scientists to substitute synthetic plastics in our daily lives. However, its industrial production is hampered by its high cost. In this study, an extensive screening program was done to isolate bacteria with high PHB productivity from agricultural fields and develop a cost-effective PHB production. A promising bacterial isolate Acinetobacter baumannii P39 was recovered and identified using 16S ribosomal gene sequencing. It produced 24% PHB per dry weight after 48 h. Several experiments were conducted to optimize the composition of the culture medium and environmental factors for the selected isolate. Results revealed that 60% aeration, 28°C incubation temperature and initial pH 7.5 showed the highest productivity. Besides, 0.7% corn oil and 0.1 g/L peptone were the best carbon and nitrogen sources, respectively. Substituting glucose with corn oil led to a 23% reduction in total input cost and an estimate price for 1kg PHB is 20.5 L.E. Strain improvement by UV mutation succeeded in improving PHB production by two fold in the selected mutant P39M2. Finally, this study valorizes usage of Acinetobacter isolate in PHB production in addition to solving the critical problem of high cost of production.

Production of poly(hydroxybutyrate-hydroxyvalerate) from waste organics by the two-stage process: focus on the intermediate volatile fatty acids.

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Shen, Liang; Hu, Hongyou; Ji, Hongfang; Cai, Jiyuan; He, Ning; Li, Qingbiao; Wang, Yuanpeng

2014-08-01

The two-stage process, coupling volatile fatty acids (VFAs) fermentation and poly(hydroxybutyrate-hydroxyvalerate) (P(HB/HV)) biosynthesis, was investigated for five waste organic materials. The overall conversion efficiencies were glycerol>starch>molasses>waste sludge>protein, meanwhile the maximum P(HB/HV) (1.674 g/L) was obtained from waste starch. Altering the waste type brought more effects on VFAs composition other than the yield in the first stage, which in turn greatly changed the yield in the second stage. Further study showed that even-number carbon VFAs (or odd-number ones) had a good positive linear relationship with P(HB/HV) content of HB (or HV). Additionally, VFA producing microbiota was analyzed by pyrosequencing methods for five wastes, which indicated that specific species (e.g., Lactobacillus for protein; Ethanoligenens for starch; Ruminococcus and Limnobacter for glycerol) were dominant in the community for VFAs production. Potential competition among acidogenic bacteria specially involved to produce some VFA was proposed as well. Copyright © 2014 Elsevier Ltd. All rights reserved.

Isolation of a bacteriophage specific for a new capsular type of Klebsiella pneumoniae and characterization of its polysaccharide depolymerase.

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Chun-Ru Hsu

Full Text Available BACKGROUND: Klebsiella pneumoniae is one of the major pathogens causing hospital-acquired multidrug-resistant infections. The capsular polysaccharide (CPS is an important virulence factor of K. pneumoniae. With 78 capsular types discovered thus far, an association between capsular type and the pathogenicity of K. pneumoniae has been observed. METHODOLOGY/PRINCIPAL FINDINGS: To investigate an initially non-typeable K. pneumoniae UTI isolate NTUH-K1790N, the cps gene region was sequenced. By NTUH-K1790N cps-PCR genotyping, serotyping and determination using a newly isolated capsular type-specific bacteriophage, we found that NTUH-K1790N and three other isolates Ca0507, Ca0421 and C1975 possessed a new capsular type, which we named KN2. Analysis of a KN2 CPS(- mutant confirmed the role of capsule as the target recognized by the antiserum and the phage. A newly described lytic phage specific for KN2 K. pneumoniae, named 0507-KN2-1, was isolated and characterized using transmission electron microscopy. Whole-genome sequencing of 0507-KN2-1 revealed a 159 991 bp double-stranded DNA genome with a G+C content of 46.7% and at least 154 open reading frames. Based on its morphological and genomic characteristics, 0507-KN2-1 was classified as a member of the Myoviridae phage family. Further analysis of this phage revealed a 3738-bp gene encoding a putative polysaccharide depolymerase. A recombinant form of this protein was produced and assayed to confirm its enzymatic activity and specificity to KN2 capsular polysaccharides. KN2 K. pneumoniae strains exhibited greater sensitivity to this depolymerase than these did to the cognate phage, as determined by spot analysis. CONCLUSIONS/SIGNIFICANCE: Here we report that a group of clinical strains possess a novel Klebsiella capsular type. We identified a KN2-specific phage and its polysaccharide depolymerase, which could be used for efficient capsular typing. The lytic phage and depolymerase also have potential as

The effect of anaerobic-aerobic and feast-famine cultivation pattern on bacterial diversity during poly-β-hydroxybutyrate production from domestic sewage sludge.

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Liu, Changli; Liu, Di; Qi, Yingjie; Zhang, Ying; Liu, Xi; Zhao, Min

2016-07-01

The main objective of this work was to investigate the influence of different oxygen supply patterns on poly-β-hydroxybutyrate (PHB) yield and bacterial community diversity. The anaerobic-aerobic (A/O) sequencing batch reactors (SBR1) and feast-famine (F/F) SBR2 were used to cultivate activated sludge to produce PHB. The mixed microbial communities were collected and analyzed after 3 months cultivation. The PHB maximum yield was 64 wt% in SBR1 and 53 wt% in SBR2. Pyrosequencing analysis 16S rRNA gene of two microbial communities indicated there were nine and four bacterial phyla in SBR1 and SBR2, respectively. Specifically, Proteobacteria (36.4 % of the total bacterial community), Actinobacteria (19.7 %), Acidobacteria (14.1 %), Firmicutes (4.4 %), Bacteroidetes (1.7 %), Cyanobacteria/Chloroplast (1.5 %), TM7 (0.8 %), Gemmatimonadetes (0.2 %), and Nitrospirae (0.1 %) were present in SBR1. Proteobacteria (94.2 %), Bacteroidetes (2.9 %), Firmicutes (1.9 %), and Actinobacteria (0.7 %) were present in SBR2. Our results indicated the SBR1 fermentation system was more stable than that of SBR2 for PHB accumulation.

Acetone production by methylobacteria.

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Thomson, A W; O'Neill, J G; Wilkinson, J F

1976-09-01

An accumulation of acetone was observed during the metabolism of ethane and products of ethane oxidation by washed suspensions of Methylosinus trichosporium OB3B. This strain possessed an acetoacetate decarboxylase and 3-hydroxybutyrate dehydrogenase, and a decline in poly-beta-hydroxybutyric acid occurred under the same conditions as acetone formation. A pathway of acetone production from poly-beta-hydroxybutyric acid via 3-hydroxybutyrate and acetoacetate was suggested.

Probing the Kinetic Anabolism of Poly-Beta-Hydroxybutyrate in Cupriavidus necator H16 Using Single-Cell Raman Spectroscopy

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Zhanhua Tao

2016-08-01

Full Text Available Poly-beta-hydroxybutyrate (PHB can be formed in large amounts in Cupriavidus necator and is important for the industrial production of biodegradable plastics. In this investigation, laser tweezers Raman spectroscopy (LTRS was used to characterize dynamic changes in PHB content—as well as in the contents of other common biomolecule—in C. necator during batch growth at both the population and single-cell levels. PHB accumulation began in the early stages of bacterial growth, and the maximum PHB production rate occurred in the early and middle exponential phases. The active biosynthesis of DNA, RNA, and proteins occurred in the lag and early exponential phases, whereas the levels of these molecules decreased continuously during the remaining fermentation process until the minimum values were reached. The PHB content inside single cells was relatively homogenous in the middle stage of fermentation; during the late growth stage, the variation in PHB levels between cells increased. In addition, bacterial cells in various growth phases could be clearly discriminated when principle component analysis was performed on the spectral data. These results suggest that LTRS is a valuable single-cell analysis tool that can provide more comprehensive information about the physiological state of a growing microbial population.

Effects of gamma irradiation on the molecular structure and mechanical properties of biodegradable polymer poly(hydroxybutyrate)

International Nuclear Information System (INIS)

Oliveira, Leticia M. de; Araujo, Elmo S.

2005-01-01

The effects of gamma irradiation ( 60 Co) on the properties of the Brazilian biodegradable polymer, Poly(hydroxybutyrate), PHB, i.e. chemical, mechanical and structural properties were investigated. PHB is a natural polyester biosynthesized by different bacteria as a form to store carbon and energy. This new biopolymer shows a great potential in the medical and pharmaceutical applications due to the biocompatibility and biodegradation capacity, since it is reabsorbed by organism without liberation of toxic substances. As it.s well known, gamma irradiation have been considered the more functional sterilization mechanism applied to medical devices. This way, it is necessary to know the effects caused by energy transfer to the polymer system. The viscosity-average molar mass (Mv) of the irradiated PHB, measured using an Ostwald-type capillary viscometer, significantly decreased. The irradiated samples (test specimens) showed a molecular degradation degree, G (scissions/100 eV) value, in the sterilization dose range (0-25 kGy) about 11.4, and 20.9 to doses above 35 kGy. Other results also indicate that the gamma irradiation significantly affected the mechanical properties of PHB. Tensile strength, impact strength and elongation at break decreased dramatically, indicating increasing on the brittleness, because significant chain scissions take place in the amorphous region of irradiated PHB. On the other hand, Young modulus does not significantly change on irradiated polymer. 13 C NMR spectra of irradiated PHB at dose of 200 kGy did not show arising of new structural groups. (author)

In vivo effect of the D-(-) isomer or natural form of 3-hydroxybutyrate on initial release of lactate dehydrogenase from the acutely ischaemic myocardium.

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Lammerant, J; Huynh-Thu, T; Kolanowski, J

1986-07-01

D-(-)-3-hydroxybutyrate, the isomer found in the circulation and in the urine of diabetic patients, generally is believed to be the physiologically important form of 3-hydroxybutyrate [10]. Little is known concerning the effects of an elevated plasma level of the D-(-) isomer of 3-hydroxybutyrate upon the acutely ischaemic heart. Using anaesthetized intact dogs with a balloon catheter inserted into the proximal part of the left anterior descending coronary artery (LAD), we have recently demonstrated that a 1 mM ketonaemia induced with the arginine salt of D-(-)-3-hydroxybutyric acid reduces the uptake of non-esterified fatty acids (NEFA) in the myocardial area distal to the inflated balloon [4]. The question arises as to whether the concomitant increase in ketone uptake in this area could be detrimental to the acutely ischaemic myocardium. Indeed, a previous study on isolated coronary ligated hearts from normal rats has shown that the rate of release of lactate dehydrogenase (LDH) during the first 90 min of ischaemia can be enhanced by replacing glucose (11 mM) in the perfusion fluid with either albumin-bound palmitate (0.9 mM) or sodium DL-3-hydroxybutyrate (10 mM) as the sole energy substrate [11]. This would suggest that the ketone might be as deleterious as its metabolic precursors for membrane integrity in the acutely ischaemic myocardium. In the present report, we examine the effect of arginine D-(-)-3-hydroxybutyrate on LDH release from ischaemic myocardium in our in vivo preparation. The dogs were treated with lidocaine in order to minimize the frequency and, hence, the adverse metabolic effects of ectopic beats.(ABSTRACT TRUNCATED AT 250 WORDS)

GC-MS based Gestational Diabetes Mellitus longitudinal study: Identification of 2-and 3-hydroxybutyrate as potential prognostic biomarkers.

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Dudzik, Danuta; Zorawski, Marcin; Skotnicki, Mariusz; Zarzycki, Wieslaw; García, Antonia; Angulo, Santiago; Lorenzo, M Paz; Barbas, Coral; Ramos, M Pilar

2017-09-10

Gestational Diabetes Mellitus (GDM) causes severe short- and long-term complications for the mother, fetus and neonate, including type 2-diabetes (T2DM) later in life. In this pilot study, GC-Q/MS analysis was applied for plasma metabolomics fingerprinting of 24 healthy and 24 women with GDM at different stages of gestation (second and third trimester) and postpartum (one and three months). Multivariate (unsupervised and supervised) statistical analysis was performed to investigate variance in the data, identify outliers and for unbiased assessment of data quality. Plasma fingerprints allowed for the discrimination of GDM pregnant women from controls both in the 2nd and 3rd trimesters of gestation. However, metabolic profiles tended to be similar after delivery. Follow up of these women revealed that 4 of them developed T2DM within 2 years postpartum. Multivariate PLS-DA models limited to women with GDM showed clear separation 3 months postpartum. In the 2nd trimester of gestation there was also a clear separation between GDM women that were normoglycemic after pregnancy and those with recognized postpartum T2DM. Metabolites that had the strongest discriminative power between these groups in the 2nd trimester of gestation were 2-hydroxybutyrate, 3-hydroxybutyrate, and stearic acid. We have described, that early GDM comprises metabotypes that are associated with the risk of future complications, including postpartum T2DM. In this pilot study, we provide evidence that 2-hydroxybutyrate and 3-hydroxybutyrate may be considered as future prognostic biomarkers to predict the onset of diabetic complications in women with gestational diabetes after delivery. Copyright © 2017 Elsevier B.V. All rights reserved.

A balanced ATP driving force module for enhancing photosynthetic biosynthesis of 3-hydroxybutyrate from CO2.

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Ku, Jason T; Lan, Ethan I

2018-03-01

Using engineered photoautotrophic microorganisms for the direct chemical synthesis from CO 2 is an attractive direction for both sustainability and CO 2 mitigation. However, the behaviors of non-native metabolic pathways may be difficult to control due to the different intracellular contexts between natural and heterologous hosts. While most metabolic engineering efforts focus on strengthening driving forces in pathway design to favor biochemical production in these organisms, excessive driving force may be detrimental to product biosynthesis due to imbalanced cellular intermediate distribution. In this study, an ATP-hydrolysis based driving force module was engineered into cyanobacterium Synechococcus elongatus PCC 7942 to produce 3-hydroxybutyrate (3HB), a valuable chemical feedstock for the synthesis of biodegradable plastics and antibiotics. However, while the ATP driving force module is effective for increasing product formation, uncontrolled accumulation of intermediate metabolites likely led to metabolic imbalance and thus to cell growth inhibition. Therefore, the ATP driving force module was reengineered by providing a reversible outlet for excessive carbon flux. Upon expression of this balanced ATP driving force module with 3HB biosynthesis, engineered strain produced 3HB with a cumulative titer of 1.2 g/L, a significant increase over the initial strain. This result highlighted the importance of pathway reversibility as an effective design strategy for balancing driving force and intermediate accumulation, thereby achieving a self-regulated control for increased net flux towards product biosynthesis. Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Effects of poly-β-hydroxybutyrate (PHB) on Siberian sturgeon (Acipenser baerii) fingerlings performance and its gastrointestinal tract microbial community.

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Najdegerami, Ebrahim H; Tran, Tiet Ngoc; Defoirdt, Tom; Marzorati, Massimo; Sorgeloos, Patrick; Boon, Nico; Bossier, Peter

2012-01-01

Poly-β-hydroxybutyrate (PHB) is a natural polymer that can be depolymerized into water-soluble short-chain fatty acid monomers. These monomers can act as microbial control agents. In this study, the effects of partially replacing the diet of Siberian sturgeon fingerlings with 2% and 5% PHB were investigated. Replacing 2% of the diet with PHB improved weight gain, specific growth rate (SGR) and survival in the sturgeon fingerlings during the 10-week experimental period. Community-level physiological profiling and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) were used to analyze the microbial community diversity and community organization in the sturgeon gastrointestinal tract. DGGE analysis revealed that PHB affected the intestinal microbial species richness and diversity. The highest species richness was observed with 2% PHB. DNA sequencing of the dominant bands in 2% and 5% PHB treatments revealed that PHB stimulated bacteria belonging to the genera Bacillus and Ruminococcaceae. Principal component analysis, Lorenz curves and the Shannon index of Biolog Ecoplate data revealed that aerobic metabolic potential of the bacterial community was different in the PHB-treated fishes as compared with the control situation. Overall, our results indicate that PHB act as microbial control agents and replacement of 2% of Siberian sturgeon fingerling diet with PHB has beneficial effects.

Effective recovery of poly-β-hydroxybutyrate (PHB) biopolymer from Cupriavidus necator using a novel and environmentally friendly solvent system.

Science.gov (United States)

Fei, Tao; Cazeneuve, Stacy; Wen, Zhiyou; Wu, Lei; Wang, Tong

2016-05-01

This work demonstrates a significant advance in bioprocessing for a high-melting lipid polymer. A novel and environmental friendly solvent mixture, acetone/ethanol/propylene carbonate (A/E/P, 1:1:1 v/v/v) was identified for extracting poly-hydroxybutyrate (PHB), a high-value biopolymer, from Cupriavidus necator. A set of solubility curves of PHB in various solvents was established. PHB recovery of 85% and purity of 92% were obtained from defatted dry biomass (DDB) using A/E/P. This solvent mixture is compatible with water, and from non-defatted wet biomass, PHB recovery of 83% and purity of 90% were achieved. Water and hexane were evaluated as anti-solvents to assist PHB precipitation, and hexane improved recovery of PHB from biomass to 92% and the purity to 93%. A scale-up extraction and separation reactor was designed, built and successfully tested. Properties of PHB recovered were not significantly affected by the extraction solvent and conditions, as shown by average molecular weight (1.4 × 10(6) ) and melting point (175.2°C) not being different from PHB extracted using chloroform. Therefore, this biorenewable solvent system was effective and versatile for extracting PHB biopolymers. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:678-685, 2016. © 2016 American Institute of Chemical Engineers.

Stenotrophomonas sp. RZS 7, a novel PHB degrader isolated from plastic contaminated soil in Shahada, Maharashtra, Western India.

Science.gov (United States)

Wani, S J; Shaikh, S S; Tabassum, B; Thakur, R; Gulati, A; Sayyed, R Z

2016-12-01

This paper reports an isolation and identification of novel poly-β-hydroxybutyrate (PHB) degrading bacterium Stenotrophomonas sp. RZS 7 and studies on its extracellular PHB degrading depolymerase enzyme. The bacterium isolated from soil samples of plastic contaminated sites of municipal area in Shahada, Maharashtra, Western India. It was identified as Stenotrophomonas sp. RZS 7 based on polyphasic approach. The bacterium grew well in minimal salt medium (MSM) and produced a zone (4.2 mm) of PHB hydrolysis on MSM containing PHB as the only source of nutrient. An optimum yield of enzyme was obtained on the fifth day of incubation at 37 °C and at pH 6.0. Further increase in enzyme production was recorded with Ca 2+ ions, while other metal ions like Fe 2+ (1 mM) and chemical viz. mercaptoethanol severally affected the production of enzyme.

Composite hydrogel of chitosan-poly(hydroxybutyrate-co-valerate) with chondroitin sulfate nanoparticles for nucleus pulposus tissue engineering.

Science.gov (United States)

Nair, Manitha B; Baranwal, Gaurav; Vijayan, Prajuna; Keyan, Kripa S; Jayakumar, R

2015-12-01

Intervertebral disc degeneration, occurring mainly in nucleus pulposus (NP), is a leading cause of low back pain. In seeking to mitigate this condition, investigators in the field of NP tissue engineering have increasingly studied the use of hydrogels. However, these hydrogels should possess appropriate mechanical strength and swelling pressure, and concurrently support the proliferation of chondrocyte-like cells. The objective of this study was to develop and validate a composite hydrogel for NP tissue engineering, made of chitosan-poly(hydroxybutyrate-co-valerate) (CP) with chondroitin sulfate (CS) nanoparticles, without using a cross linker. The water uptake ability, as well as the viscoelastic properties of this composite hydrogel, was similar to native tissue, as reflected in the complex shear modulus and stress relaxation values. The hydrogel could withstand varying stress corresponding to daily activities like lying down (0.01 MPa), sitting (0.5 MPa) and standing (1.0 MPa) under dynamic conditions. The hydrogels were stable in PBS for 2 weeks and its stiffness, elastic and viscous modulus did not alter significantly during this period. Both CP and CP-CS hydrogels could assist the viability and adhesion of adipose derived rat mesenchymal stem cells (ADMSCs). The viability and chondrogenic differentiation of MSCs was significantly enhanced in presence of CS nanoparticles. Thus, CS nanoparticles-incorporated chitosan-PHBV hydrogels offer great potential for NP tissue engineering. Copyright © 2015 Elsevier B.V. All rights reserved.

Production of poly-β-hydroxybutyrate (PHB) by Methylobacterium organophilum isolated from a methanotrophic consortium in a two-phase partition bioreactor

International Nuclear Information System (INIS)

Zuniga, C.; Morales, M.; Le Borgne, S.; Revah, S.

2011-01-01

The biodegradation of methane, a greenhouse gas, and the accumulation of poly-β-hydroxybutyrate (PHB) were studied using a methanotrophic consortium and an isolated strain thereof. The specific rates for methane consumption were 100 and 17mg CH 4 g x -1 h -1 for the isolate and the consortium, respectively. Also the effect of including 10% (v v -1 ) of silicone oil in a two-phase partitioning bioreactor (TPPB) was assayed for the elimination of 1% methane in air stream. TPPB allowed a 33-45% increase of methane elimination under growing conditions. Nitrogen limitation was assayed in bioreactors to promote PHB production. Under this condition, the specific methane degradation rate remained unchanged for the consortium and decreased to 36mg CH 4 g x -1 h -1 for the isolated strain. The accumulated PHB in the reactor was 34% and 38% (w w -1 ) for the consortium and the isolate, respectively. The highest productivity was obtained in the TPPB and was 1.61mg PHB g x -1 h -1 . The CZ-2 isolate was identified as Methylobacterium organophilum, this is the first study that reports this species as being able to grow on methane and accumulate up to 57% (w w -1 ) of PHB under nitrogen limitation in microcosm experiments.

Generation of poly-β-hydroxybutyrate from acetate in higher plants: Detection of acetoacetyl CoA reductase- and PHB synthase- activities in rice.

Science.gov (United States)

Tsuda, Hirohisa; Shiraki, Mari; Inoue, Eri; Saito, Terumi

2016-08-20

It has been reported that Poly-β-hydroxybutyrate (PHB) is generated from acetate in the rice root. However, no information is available about the biosynthetic pathway of PHB from acetate in plant cells. In the bacterium Ralstonia eutropha H16 (R. eutropha), PHB is synthesized from acetyl CoA by the consecutive reaction of three enzymes: β-ketothiolase (EC: 2.3.1.9), acetoacetyl CoA reductase (EC: 1.1.1.36) and PHB synthase (EC: 2.3.1.-). Thus, in this study, we examined whether the above three enzymatic activities were also detected in rice seedlings. The results clearly showed that the activities of the above three enzymes were all detected in rice. In particular, the PHB synthase activity was detected specifically in the sonicated particulate fractions (2000g 10min precipitate (ppt) and the 8000g 30min ppt) of rice roots and leaves. In addition to these enzyme activities, several new experimental results were obtained on PHB synthesis in higher plants: (a) (14)C-PHB generated from 2-(14)C-acetate was mainly localized in the 2000g 10min ppt and the 8000g 30min ppt of rice root. (b) Addition of acetate (0.1-10mM) to culture medium of rice seedlings did not increase the content of PHB in the rice root or leaf. (c) In addition to C3 plants, PHB was generated from acetate in a C4 plant (corn) and in a CAM plant (Bryophyllum pinnatum). d) Washing with ethylenediaminetetraacetic acid (EDTA) strongly suggested that the PHB synthesized from acetate was of plant origin and was not bacterial contamination. Copyright © 2016 Elsevier GmbH. All rights reserved.

Poly-ε-caprolactone Coated and Functionalized Porous Titanium and Magnesium Implants for Enhancing Angiogenesis in Critically Sized Bone Defects.

Science.gov (United States)

Roland, Laura; Grau, Michael; Matena, Julia; Teske, Michael; Gieseke, Matthias; Kampmann, Andreas; Beyerbach, Martin; Murua Escobar, Hugo; Haferkamp, Heinz; Gellrich, Nils-Claudius; Nolte, Ingo

2015-12-22

For healing of critically sized bone defects, biocompatible and angiogenesis supporting implants are favorable. Murine osteoblasts showed equal proliferation behavior on the polymers poly-ε-caprolactone (PCL) and poly-(3-hydroxybutyrate)/poly-(4-hydroxybutyrate) (P(3HB)/P(4HB)). As vitality was significantly better for PCL, it was chosen as a suitable coating material for further experiments. Titanium implants with 600 µm pore size were evaluated and found to be a good implant material for bone, as primary osteoblasts showed a vitality and proliferation onto the implants comparable to well bottom (WB). Pure porous titanium implants and PCL coated porous titanium implants were compared using Live Cell Imaging (LCI) with Green fluorescent protein (GFP)-osteoblasts. Cell count and cell covered area did not differ between the implants after seven days. To improve ingrowth of blood vessels into porous implants, proangiogenic factors like Vascular Endothelial Growth Factor (VEGF) and High Mobility Group Box 1 (HMGB1) were incorporated into PCL coated, porous titanium and magnesium implants. An angiogenesis assay was performed to establish an in vitro method for evaluating the impact of metallic implants on angiogenesis to reduce and refine animal experiments in future. Incorporated concentrations of proangiogenic factors were probably too low, as they did not lead to any effect. Magnesium implants did not yield evaluable results, as they led to pH increase and subsequent cell death.

Marine fungi: Degraders of poly-3-hydroxyalkanoate based plastic materials

Directory of Open Access Journals (Sweden)

Matavulj Milan

2009-01-01

Full Text Available The search for new biosynthetic and biodegradable materials to save nonrenewable resources and reduce global pollution problems is an urgent task. Recently, materials like thermoplastic poly-3-hydroxyalkanoates (PHA, have been found synthesized by bacteria as storage materials. The major PHAs synthesized are poly-b-hydroxybutyrate (PHB, poly-b-hydroxyvalerate (PHV and their copolymers. They are already commercially produced and used as BIOPOLTM (ICI, England. Their complete degradability by bacteria has already been shown. Today, oceans and estuaries serve as major landfills, and since fungi are an important part of the degrading microbiota, in order to prove their participation in the degradation process, a simple degradation test suitable for fungi and marine conditions had to be developed. Several solid media based on artificial sea water, differing in the content of non-alkanoate organics and supplemented with 0.1% PHA (or BIOPOLTM as a main source of carbon have been tested. The testing principle consists of clearing the turbid medium in test tube or plates caused by suspended granules of PHA. All media tested supported the growth of fungi. For the discrete and transparent clearing of zones, a mineral medium with 0.01% peptone, 0.01% yeast extract, and 0.1% PHB or BIOPOLTM was finally chosen where the fine and evenly distributed turbidity is accomplished by a specific procedure. This method allows the investigation of degradability of PHA-based plastic materials as well as screening for fungal ability to depolymerise pure PHA homopolymers. Using this medium, 32 strains of marine yeasts and 102 strains of marine mycelial fungi belonging to different systematic and ecological groups were tested for their ability to degrade PHAs. Only about 4% of the strains were able to degrade BIOPOLTM and about 6% depolymerised pure PHB homopolymer. This is in sharp contrast to the results of our previous experiments with 143 strains of terrestrial fungi

Synthesis and aqueous phase behavior of thermoresponsive biodegradable poly(D,L-3-methylglycolide)-block-poly(ethyelene glycol)-block-poly(D,L-3-methylglycolide) triblock copolymers

NARCIS (Netherlands)

Zhong, Zhiyuan; Dijkstra, Pieter J.; Feijen, Jan; Kwon, Young-Min; Bae, You Han; Kim, Sung Wan

2002-01-01

Novel biodegradable thermosensitive triblock copolymers of poly(D,L-3-methylglycolide)-block-poly(ethylene glycol)-block-poly(D,L-3-methylglycolide) (PMG-PEG-PMG) have been synthesized. Ring-opening polymerization of D,L-3-methyl-glycolide (MG) initiated with poly(ethylene glycol) (PEG) and

The Use of Poly-4-Hydroxybutyrate (P4HB) Scaffold in the Ptotic Breast: A Multicenter Clinical Study.

Science.gov (United States)

Adams, William P; Baxter, Richard; Glicksman, Caroline; Mast, Bruce A; Tantillo, Michael; Van Natta, Bruce W

2018-04-06

Mastopexy and reduction mammaplasty are often limited by the patient's poor native soft tissue quality, resulting in ptosis recurrence and loss of rejuvenated surgical results. Surgical scaffolds and acellular dermal matrices are used in these procedures to provide physical and mechanical stabilization of weakened or compromised tissue. GalaFLEX scaffold, made from poly-4-hydroxybutyrate (P4HB), is a next-generation product for soft tissue reinforcement that resorbs gradually while aiding tissue regeneration to achieve excellent outcomes. To assess the clinical performance of GalaFLEX scaffold in soft tissue reinforcement during elective mastopexy and reduction mammaplasty. This multicenter, single-arm, observational study assessed product performance and outcomes of GalaFLEX scaffold when used in breast surgery. Outcomes included ptosis correction and maintenance, associated adverse events, patient and surgeon satisfaction, and mammographic and ultrasound imaging evaluation. At 6 centers in the US, 62 of 69 enrolled patients were treated. Of this population, 89.7% had successful ptosis correction and maintenance at 1 year, with high patient and surgeon satisfaction for breast shape, droop/sag of the breast, and maintenance of results at 1 year. There were 5 adverse events deemed related to the device (8.0%), including nerve pain, breast swelling, ptosis, and 2 instances of asymmetry. GalaFLEX scaffold safely and successfully supports and elevates breast tissue in mastopexy and reduction mammaplasty, with maintained support at 1 year. Surgeon and patient satisfaction were high. No mammogram or ultrasound interference was detected.

Development of curcumin-loaded poly(hydroxybutyrate- co-hydroxyvalerate) nanoparticles as anti-inflammatory carriers to human-activated endothelial cells

Science.gov (United States)

Simion, Viorel; Stan, Daniela; Gan, Ana-Maria; Pirvulescu, Monica Madalina; Butoi, Elena; Manduteanu, Ileana; Deleanu, Mariana; Andrei, Eugen; Durdureanu-Angheluta, Anamaria; Bota, Marian; Enachescu, Marius; Calin, Manuela; Simionescu, Maya

2013-12-01

Curcumin (Cm)-loaded poly(hydroxybutyrate- co-hydroxyvalerate) (PHBV) nanoparticles (CmPN) were obtained and characterized and their effect on human endothelial cells (HEC) was assessed. Different CmPN formulations have been prepared using the emulsion solvent evaporation technique, and characterized for size, structure, Zeta potential, Cm entrapment efficiency, and in vitro Cm release. CmPN cytotoxicity and cellular uptake have been followed using HEC. Also, the effect of CmPN treatment on the p38MAPK signaling pathway in endothelial cells was investigated. The results obtained by electron and atomic force microscopy revealed the spherical shape of the CmPN formulation. Based on size and encapsulation efficiency, the CmPN formulation with the average diameter of 186 nm and with the highest encapsulation efficiency (83 %) has been used in the further studies. The release of Cm from CmPN was 18 % after 8 h of incubation at 37 °C, followed by a slow release until 144 h, when it reached 44 %, indicating a controlled release. CmPN are taken up by HEC and exhibited low cytotoxicity at concentrations up to 10 μM. The pre-treatment of HEC with CmPN before exposure to tumor necrosis factor-alpha (TNF-α) determined a decrease of p38MAPK phosphorylation. In conclusion, Cm encapsulated into PHBV nanoparticles, at concentration up to 10 μM, has low cytotoxicity and display anti-inflammatory activity on TNF-α-activated HEC by suppressing the phosphorylation of p38MAPK.

Functional characterization of salt-tolerant microbial esterase WDEst17 and its use in the generation of optically pure ethyl (R)-3-hydroxybutyrate.

Science.gov (United States)

Wang, Yilong; Xu, Yongkai; Zhang, Yun; Sun, Aijun; Hu, Yunfeng

2018-06-01

The two enantiomers of ethyl 3-hydroxybutyrate are important intermediates for the synthesis of a great variety of valuable chiral drugs. The preparation of chiral drug intermediates through kinetic resolution reactions catalyzed by esterases/lipases has been demonstrated to be an efficient and environmentally friendly method. We previously functionally characterized microbial esterase PHE21 and used PHE21 as a biocatalyst to generate optically pure ethyl (S)-3-hydroxybutyrate. Herein, we also functionally characterized one novel salt-tolerant microbial esterase WDEst17 from the genome of Dactylosporangium aurantiacum subsp. Hamdenensis NRRL 18085. Esterase WDEst17 was further developed as an efficient biocatalyst to generate (R)-3-hydroxybutyrate, an important chiral drug intermediate, with the enantiomeric excess being 99% and the conversion rate being 65.05%, respectively, after process optimization. Notably, the enantio-selectivity of esterase WDEst17 was opposite than that of esterase PHE21. The identification of esterases WDEst17 and PHE21 through genome mining of microorganisms provides useful biocatalysts for the preparation of valuable chiral drug intermediates. © 2018 Wiley Periodicals, Inc.

Specific gamma-hydroxybutyrate-binding sites but loss of pharmacological effects of gamma-hydroxybutyrate in GABA-B1-deficient mice

DEFF Research Database (Denmark)

Kaupmann, Klemens; Cryan, John F; Wellendorph, Petrine

2003-01-01

gamma-Hydroxybutyrate (GHB), a metabolite of gamma-aminobutyric acid (GABA), is proposed to function as a neurotransmitter or neuromodulator. gamma-Hydroxybutyrate and its prodrug, gamma-butyrolactone (GBL), recently received increased public attention as they emerged as popular drugs of abuse. T...

Nanoparticles of poly(hydroxybutyrate-co-hydroxyvalerate) as support for the immobilization of Candida antarctica lipase (fraction B); Nanoparticulas de poli-hidroxibutirato-co-valerato como suporte para a imobilizacao da lipase de Candida antarctica fracao B

Energy Technology Data Exchange (ETDEWEB)

Fernandes, Ilizandra A.; Nyari, Nadia L.D. [Universidade Regional Integrada, Erechim, RS (Brazil). Departamento de Ciencias Agrarias; Oliveira, Jose Vladimir de; Oliveira, Debora de, E-mail: debora@enq.ufsc.br [Universidade Federal de Santa Catarina (UFSC), Florianopolis, SC (Brazil). Departamento de Engenharia Quimica e Engenharia de Alimentos; Rigo, Elisandra [Universidade do Estado de Santa Catarina (UDESC), Pinhalzinho, SC (Brazil). Departamento de Engenharia de Alimentos; Souza, Maria Cristiane M. de; Goncalves, Luciana R.B. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil). Departamento de Engenharia Quimica; Pergher, Sibele Berenice C. [Universidade Federal do Rio Grande do Norte (UFRN), RN (Brazil). Instituto de Quimica

2014-04-15

This work evaluates the immobilization of Candida antarctica lipase (Fraction B) using poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) nanoparticles as support. The effects of immobilization time (30-150 min) and pH (5-10) on lipase loading were evaluated. The stability of the immobilized enzyme towards temperature (40, 60, and 80 deg C), reuse and storage (at 4 deg C) were also determined. Furthermore, to assess its potential application in a system of interest, the immobilized lipase was used as a catalyst in the esterification of geraniol with oleic acid. The results indicated a time of 120 minutes and pH of 7 as optimal for immobilization. A 21 hour exposure of the PHBV-lipase derivative to 60 deg C showed a 33% reduction of the initial activity while storage at 4 deg C led to a residual activity (5% of the original activity). The derivative was used without significant loss of activity for 4 successive cycles. The use of the immobilized lipase as a catalyst in the production of geranyl oleate led to about 88% conversion of the initial reactants to products. (author)

Production of poly-{beta}-hydroxybutyrate (PHB) by Methylobacterium organophilum isolated from a methanotrophic consortium in a two-phase partition bioreactor

Energy Technology Data Exchange (ETDEWEB)

Zuniga, C., E-mail: cristal7n@gmail.com [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico); Morales, M., E-mail: mmorales@correo.cua.uam.mx [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico); Le Borgne, S., E-mail: sylvielb@correo.cua.uam.mx [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico); Revah, S., E-mail: srevah@correo.cua.uam.mx [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico)

2011-06-15

The biodegradation of methane, a greenhouse gas, and the accumulation of poly-{beta}-hydroxybutyrate (PHB) were studied using a methanotrophic consortium and an isolated strain thereof. The specific rates for methane consumption were 100 and 17mg{sub CH{sub 4}}g{sub x}{sup -1} h{sup -1} for the isolate and the consortium, respectively. Also the effect of including 10% (v v{sup -1}) of silicone oil in a two-phase partitioning bioreactor (TPPB) was assayed for the elimination of 1% methane in air stream. TPPB allowed a 33-45% increase of methane elimination under growing conditions. Nitrogen limitation was assayed in bioreactors to promote PHB production. Under this condition, the specific methane degradation rate remained unchanged for the consortium and decreased to 36mg{sub CH{sub 4}}g{sub x}{sup -1} h{sup -1} for the isolated strain. The accumulated PHB in the reactor was 34% and 38% (w w{sup -1}) for the consortium and the isolate, respectively. The highest productivity was obtained in the TPPB and was 1.61mg{sub PHB}g{sub x}{sup -1} h{sup -1}. The CZ-2 isolate was identified as Methylobacterium organophilum, this is the first study that reports this species as being able to grow on methane and accumulate up to 57% (w w{sup -1}) of PHB under nitrogen limitation in microcosm experiments.

Growth of Azotobacter vinelandii UWD in Fish Peptone Medium and Simplified Extraction of Poly-β-Hydroxybutyrate

Science.gov (United States)

Page, William J.; Cornish, Anthony

1993-01-01

Azotobacter vinelandii UWD was grown in a fermentor with glucose medium with and without 0.1% fish peptone (FP) in batch and fed-batch cultures for the production of the natural bioplastic poly-β-hydroxybutyrate (PHB). Strain UWD formed PHB five times faster than cell protein during growth in glucose and NH4+, but PHB synthesis stopped when NH4+ was depleted and nitrogen fixation started. When FP was added to the same medium, PHB accumulated 16 times faster than cell protein, which in turn was inhibited by 40%, and PHB synthesis was unaffected by NH4+ depletion. Thus, FP appeared to be used as a nitrogen source by these nitrogen-fixing cells, which permitted enhanced PHB synthesis, but it was not a general growth stimulator. The addition of FP to the medium led to the production of large, pleomorphic, osmotically sensitive cells that demonstrated impaired growth and partial lysis, with the leakage of DNA into the culture fluid, but these cells were still able to synthesize PHB at elevated rates and efficiency. When FP was continuously present in fed-batch culture, the yield in grams of polymer per gram of glucose consumed was calculated to range from 0.43 g/g, characteristic of nongrowing cells, to an unprecedented 0.65 g/g. Separation of an FP-free growth phase from an FP-containing growth phase in fed-batch culture resulted in better growth of these pleomorphic cells and good production of PHB (yield, 0.32 g/g). The fragility of these cells was exploited in a simple procedure for the extraction of high-molecular-weight PHB. The cells were treated with 1 N aqueous NH3 (pH 11.4) at 45°C for 10 min. This treatment removed about 10% of the non-PHB mass from the pellet, of which 60 to 77% was protein. The final product consisted of 94% PHB, 2% protein, and 4% nonprotein residual mass. The polymer molecular weight (1.7 × 106 to 2.0 × 106) and dispersity (1.0 to 1.9) were not significantly affected (P = 0.05) by this treatment. In addition, the NH3 extraction waste

The Opportunity for High-Performance Biomaterials from Methane.

Science.gov (United States)

Strong, Peter James; Laycock, Bronwyn; Mahamud, Syarifah Nuraqmar Syed; Jensen, Paul Douglas; Lant, Paul Andrew; Tyson, Gene; Pratt, Steven

2016-02-03

Polyhydroxyalkanoate (PHA) biopolymers are widely recognised as outstanding candidates to replace conventional petroleum-derived polymers. Their mechanical properties are good and can be tailored through copolymer composition, they are biodegradable, and unlike many alternatives, they do not rely on oil-based feedstocks. Further, they are the only commodity polymer that can be synthesised intracellularly, ensuring stereoregularity and high molecular weight. However, despite offering enormous potential for many years, they are still not making a significant impact. This is broadly because commercial uptake has been limited by variable performance (inconsistent polymer properties) and high production costs of the raw polymer. Additionally, the main type of PHA produced naturally is poly-3-hydroxybutyrate (PHB), which has limited scope due to its brittle nature and low thermal stability, as well as its tendency to embrittle over time. Production cost is strongly impacted by the type of the feedstock used. In this article we consider: the production of PHAs from methanotrophs using methane as a cost-effective substrate; the use of mixed cultures, as opposed to pure strains; and strategies to generate a poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer (PHBV), which has more desirable qualities such as toughness and elasticity.

Blood glucose, lactate, pyruvate, glycerol, 3-hydroxybutyrate and acetoacetate measurements in man using a centrifugal analyser with a fluorimetric attachment.

Science.gov (United States)

Harrison, J; Hodson, A W; Skillen, A W; Stappenbeck, R; Agius, L; Alberti, K G

1988-03-01

Methods are described for the analysis of glucose, lactate, pyruvate, alanine, glycerol, 3-hydroxybutyrate and acetoacetate in perchloric acid extracts of human blood, using the Cobas Bio centrifugal analyser fitted with a fluorimetric attachment. Intra-assay and inter-assay coefficients of variation ranged from 1.9 to 7.9% and from 1.0 to 7.2% respectively. Correlation coefficients ranged from 0.96 to 0.99 against established continuous-flow and manual spectrophotometric methods. All seven metabolites can be measured using a single perchloric acid extract of 20 microliter of blood. The versatility of the assays is such that as little as 100 pmol pyruvate, 3-hydroxybutyrate or as much as 15 nmol glucose can be measured in the same 20 microliter extract.

Preparo e caracterização de micropartículas de acetobutirato de celulose e poli(3-hidroxibutirato contendo piroxicam - DOI: 10.4025/actascihealthsci.v31i1.4428 Preparation and characterization of cellulose acetate butyrate and poly(3-hydroxybutyrate microparticles containing piroxicam - DOI: 10.4025/actascihealthsci.v31i1.4428

Directory of Open Access Journals (Sweden)

Melissa Zétola

2009-05-01

Full Text Available O objetivo deste trabalho foi avaliar a influência da massa molar do acetobutirato de celulose (ABC e da adição de poli(3-hidroxibutirato [PHB] sobre a morfologia das micropartículas, a eficiência de encapsulação e os perfis de liberação do piroxicam. As micropartículas foram preparadas por meio da técnica de emulsão/evaporação do solvente O/A e caracterizadas quanto à morfologia por microscopia eletrônica de varredura. O teor de fármaco nas micropartículas foi determinado utilizando o método de espectrofotometria de absorção na região do ultravioleta; os ensaios de liberação realizados, utilizando tampão fosfato pH 6,8. As micropartículas obtidas apresentaram formas irregulares, e aquelas preparadas a partir do ABC com maior massa molar apresentaram maior tamanho. Mediante planejamento fatorial, observou-se que as variáveis analisadas (massa molar do ABC e adição de PHB não influenciaram a eficiência de encapsulação do piroxicam, mas exerceram influência sobre a quantidade inicial de piroxicam liberada a partir das micropartículas.This work aims to evaluate the influence of the cellulose acetate butyrate (CAB molar weight and the addition of poly(3-hydroxybutyrate [PHB] on microparticle morphology, encapsulation efficiency and release profile of piroxicam. The microparticles were prepared using the O/W emulsion/solvent evaporation technique and characterized according to the morphology using scanning electron microscopy. The drug content in the microparticles was determined through UV spectrophotometry and a dissolution assay was conducted using phosphate buffer pH 6.8. The obtained microparticles presented irregular shape; the ones prepared with CAB with large molar weight presented a larger size. Through a factorial design, it was observed that the analyzed variables (CAB molar weight and PHB addition did not influence the encapsulation efficiency, but did influence the initial release of piroxicam from the

Poly-ε-caprolactone Coated and Functionalized Porous Titanium and Magnesium Implants for Enhancing Angiogenesis in Critically Sized Bone Defects

Directory of Open Access Journals (Sweden)

Laura Roland

2015-12-01

Full Text Available For healing of critically sized bone defects, biocompatible and angiogenesis supporting implants are favorable. Murine osteoblasts showed equal proliferation behavior on the polymers poly-ε-caprolactone (PCL and poly-(3-hydroxybutyrate/poly-(4-hydroxybutyrate (P(3HB/P(4HB. As vitality was significantly better for PCL, it was chosen as a suitable coating material for further experiments. Titanium implants with 600 µm pore size were evaluated and found to be a good implant material for bone, as primary osteoblasts showed a vitality and proliferation onto the implants comparable to well bottom (WB. Pure porous titanium implants and PCL coated porous titanium implants were compared using Live Cell Imaging (LCI with Green fluorescent protein (GFP-osteoblasts. Cell count and cell covered area did not differ between the implants after seven days. To improve ingrowth of blood vessels into porous implants, proangiogenic factors like Vascular Endothelial Growth Factor (VEGF and High Mobility Group Box 1 (HMGB1 were incorporated into PCL coated, porous titanium and magnesium implants. An angiogenesis assay was performed to establish an in vitro method for evaluating the impact of metallic implants on angiogenesis to reduce and refine animal experiments in future. Incorporated concentrations of proangiogenic factors were probably too low, as they did not lead to any effect. Magnesium implants did not yield evaluable results, as they led to pH increase and subsequent cell death.

(3-Aminopropyl)-4-methylpiperazine End-capped Poly(1,4-butanediol diacrylate-co-4-amino-1-butanol)-based Multilayer Films for Gene Delivery

Science.gov (United States)

Li, Cuicui; Tzeng, Stephany Y; Tellier, Liane E.; Green, Jordan J

2013-01-01

Biodegradable polyelectrolyte surfaces for gene delivery were created through electrospinning of biodegradable polycations combined with iterative solution-based multilayer coating. Poly(β-amino ester) (PBAE) poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) end-capped with 1-(3-aminopropyl)-4-methylpiperazine was utilized due to its ability to electrostatically interact with anionic molecules like DNA, its biodegradability, and its low cytotoxicity. A new DNA release system was developed for sustained release of DNA over 24 hours, accompanied by high exogenous gene expression in primary human glioblastoma (GB) cells. Electrospinning a different PBAE, poly(1,4-butanediol diacrylate-co-4,4′-trimethylenedipiperidine), and its combination with polyelectrolyte 1-(3-aminopropyl)-4-methylpiperazine end-capped poly(1,4-butanediol diacrylate-co-4-amino-1-butanol)-based multilayers are promising for DNA release and intracellular delivery from a surface. PMID:23755861

(3-aminopropyl)-4-methylpiperazine end-capped poly(1,4-butanediol diacrylate-co-4-amino-1-butanol)-based multilayer films for gene delivery.

Science.gov (United States)

Li, Cuicui; Tzeng, Stephany Y; Tellier, Liane E; Green, Jordan J

2013-07-10

Biodegradable polyelectrolyte surfaces for gene delivery were created through electrospinning of biodegradable polycations combined with iterative solution-based multilayer coating. Poly(β-amino ester) (PBAE) poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) end-capped with 1-(3-aminopropyl)-4-methylpiperazine was utilized because of its ability to electrostatically interact with anionic molecules like DNA, its biodegradability, and its low cytotoxicity. A new DNA release system was developed for sustained release of DNA over 24 h, accompanied by high exogenous gene expression in primary human glioblastoma (GB) cells. Electrospinning a different PBAE, poly(1,4-butanediol diacrylate-co-4,4'-trimethylenedipiperidine), and its combination with polyelectrolyte 1-(3-aminopropyl)-4-methylpiperazine end-capped poly(1,4-butanediol diacrylate-co-4-amino-1-butanol)-based multilayers are promising for DNA release and intracellular delivery from a surface.

Designing of Collagen Based Poly(3-hydroxybutyrate-co-4-hydroxybutyrate Scaffolds for Tissue Engineering

Directory of Open Access Journals (Sweden)

S. Vigneswari

2015-01-01

Full Text Available P(3HB-co-4HB copolymer was modified using collagen by adapting dual solvent system. The surface properties of samples were characterized by Fourier transform infrared spectroscopy (FTIR, scanning electron microscopy (SEM, organic elemental analysis (CHN analysis, and water contact angle measurements. The effects of collagen concentration, scaffold thickness, and 4HB molar fraction on the hydrophilicity were optimized by the Taguchi method. The orthogonal array experiment was conducted to obtain the response for a hydrophilic scaffold. Analysis of variance (ANOVA was used to determine the significant parameters and determine the optimal level for each parameter. The results also showed that the hydrophilicity of P(3HB-co-4HB/collagen blend scaffolds increased as the collagen concentration increased up to 15 wt% with a molar fraction of 50 mol% at 0.1 mm scaffold thickness. The biocompatibility of the P(3HB-co-4HB/collagen blend surface was evaluated by fibroblast cell (L929 culture. The collagen blend scaffold surfaces showed significant cell adhesion and growth as compared to P(3HB-co-4HB copolymer scaffolds.

rRNA and Poly-β-Hydroxybutyrate Dynamics in Bioreactors Subjected to Feast and Famine Cycles

Science.gov (United States)

Frigon, Dominic; Muyzer, Gerard; van Loosdrecht, Mark; Raskin, Lutgarde

2006-01-01

Feast and famine cycles are common in activated sludge wastewater treatment systems, and they select for bacteria that accumulate storage compounds, such as poly-β-hydroxybutyrate (PHB). Previous studies have shown that variations in influent substrate concentrations force bacteria to accumulate high levels of rRNA compared to the levels in bacteria grown in chemostats. Therefore, it can be hypothesized that bacteria accumulate more rRNA when they are subjected to feast and famine cycles. However, PHB-accumulating bacteria can form biomass (grow) throughout a feast and famine cycle and thus have a lower peak biomass formation rate during the cycle. Consequently, PHB-accumulating bacteria may accumulate less rRNA when they are subjected to feast and famine cycles than bacteria that are not capable of PHB accumulation. These hypotheses were tested with Wautersia eutropha H16 (wild type) and W. eutropha PHB-4 (a mutant not capable of accumulating PHB) grown in chemostat and semibatch reactors. For both strains, the cellular RNA level was higher when the organism was grown in semibatch reactors than when it was grown in chemostats, and the specific biomass formation rates during the feast phase were linearly related to the cellular RNA levels for cultures. Although the two strains exhibited maximum uptake rates when they were grown in semibatch reactors, the wild-type strain responded much more rapidly to the addition of fresh medium than the mutant responded. Furthermore, the chemostat-grown mutant culture was unable to exhibit maximum substrate uptake rates when it was subjected to pulse-wise addition of fresh medium. These data show that the ability to accumulate PHB does not prevent bacteria from accumulating high levels of rRNA when they are subjected to feast and famine cycles. Our results also demonstrate that the ability to accumulate PHB makes the bacteria more responsive to sudden increases in substrate concentrations, which explains their ecological

rRNA and poly-beta-hydroxybutyrate dynamics in bioreactors subjected to feast and famine cycles.

Science.gov (United States)

Frigon, Dominic; Muyzer, Gerard; van Loosdrecht, Mark; Raskin, Lutgarde

2006-04-01

Feast and famine cycles are common in activated sludge wastewater treatment systems, and they select for bacteria that accumulate storage compounds, such as poly-beta-hydroxybutyrate (PHB). Previous studies have shown that variations in influent substrate concentrations force bacteria to accumulate high levels of rRNA compared to the levels in bacteria grown in chemostats. Therefore, it can be hypothesized that bacteria accumulate more rRNA when they are subjected to feast and famine cycles. However, PHB-accumulating bacteria can form biomass (grow) throughout a feast and famine cycle and thus have a lower peak biomass formation rate during the cycle. Consequently, PHB-accumulating bacteria may accumulate less rRNA when they are subjected to feast and famine cycles than bacteria that are not capable of PHB accumulation. These hypotheses were tested with Wautersia eutropha H16 (wild type) and W. eutropha PHB-4 (a mutant not capable of accumulating PHB) grown in chemostat and semibatch reactors. For both strains, the cellular RNA level was higher when the organism was grown in semibatch reactors than when it was grown in chemostats, and the specific biomass formation rates during the feast phase were linearly related to the cellular RNA levels for cultures. Although the two strains exhibited maximum uptake rates when they were grown in semibatch reactors, the wild-type strain responded much more rapidly to the addition of fresh medium than the mutant responded. Furthermore, the chemostat-grown mutant culture was unable to exhibit maximum substrate uptake rates when it was subjected to pulse-wise addition of fresh medium. These data show that the ability to accumulate PHB does not prevent bacteria from accumulating high levels of rRNA when they are subjected to feast and famine cycles. Our results also demonstrate that the ability to accumulate PHB makes the bacteria more responsive to sudden increases in substrate concentrations, which explains their ecological

Burkholderia sacchari DSM 17165: A source of compositionally-tunable block-copolymeric short-chain poly(hydroxyalkanoates) from xylose and levulinic acid.

Science.gov (United States)

Ashby, Richard D; Solaiman, Daniel K Y; Nuñez, Alberto; Strahan, Gary D; Johnston, David B

2018-04-01

Burkholderia sacchari was used to produce poly-3-hydroxybutyrate-co-3-hydroxyvalerate block copolymers from xylose and levulinic acid. Levulinic acid was the preferred substrate resulting in 3-hydroxyvalerate (3HV) contents as high as 95 mol% at 24 h. The 3HB:3HV ratios were controlled by the initial levulinic acid media concentration and fermentation length. Higher levulinic acid concentrations and longer durations, resulted in polymers with two glass transition temperatures, each approximating those associated with poly-3HB and poly-3HV. 13 C NMR confirmed the presence of high concentrations of 3HB-3HB and 3HV-3HV homopolymeric dyads, while mass spectrometry of the partial hydrolysis products did not conform to Bernoullian statistics for randomness, confirming block sequences. MS/MS analysis of specific oligomers showed the mass-loss of 86 amu (a 3HB unit) and 100 amu (a 3HV unit) attesting to some randomness within the polymers. This study verifies the potential for producing Poly-3HB-block-3HV copolymers from inexpensive biorenewable feedstocks without sequential addition of carbon sources. Published by Elsevier Ltd.

Effect of light-dark cycles on hydrogen and poly-β-hydroxybutyrate production by a photoheterotrophic culture and Rhodobacter capsulatus using a dark fermentation effluent as substrate.

Science.gov (United States)

Montiel Corona, Virginia; Le Borgne, Sylvie; Revah, Sergio; Morales, Marcia

2017-02-01

A Rhodobacter capsulatus strain and a photoheterotrophic culture (IZT) were cultivated to produce hydrogen under different light-dark cycles. A dark fermentation effluent (DFE) was used as substrate. It was found that IZT culture had an average cumulative hydrogen production (Paccum H 2 ) of 1300±43mLH 2 L -1 under continuous illumination and light-dark cycles of 30 or 60min. In contrast, R. capsulatus reduced its Paccum H 2 by 20% under 30:30min light-dark cycles, but tripled its poly-β-hydroxybutyrate (PHB) content (308±2mgPHB gdw -1 ) compared to continuous illumination. The highest PHB content by IZT culture was 178±10mgPHB gdw -1 under 15:15min light-dark cycles. PCR-DGGE analysis revealed that the IZT culture was mainly composed of Rhodopseudomonas palustris identified with high nucleotide similarity (99%). The evaluated cultures might be used for hydrogen and PHB production. They might provide energy savings by using light-dark cycles and DFE valorization. Copyright © 2016 Elsevier Ltd. All rights reserved.

Neuronal uptake and intracellular superoxide scavenging of a fullerene (C60)-poly(2-oxazoline)s nanoformulation

KAUST Repository

Tong, Jing

2011-05-01

Fullerene, the third allotrope of carbon, has been referred to as a "radical sponge" because of its powerful radical scavenging activities. However, the hydrophobicity and toxicity associated with fullerene limits its application as a therapeutic antioxidant. In the present study, we sought to overcome these limitations by generating water-soluble nanoformulations of fullerene (C(60)). Fullerene (C(60)) was formulated with poly(N-vinyl pyrrolidine) (PVP) or poly(2-alkyl-2-oxazoline)s (POx) homopolymer and random copolymer to form nano-complexes. These C(60)-polymer complexes were characterized by UV-vis spectroscopy, infrared spectroscopy (IR), dynamic light scattering (DLS), atomic force microscopy (AFM) and transmission electron microscopy (TEM). Cellular uptake and intracellular distribution of the selected formulations in catecholaminergic (CATH.a) neurons were examined by UV-vis spectroscopy, immunofluorescence and immunogold labeling. Electron paramagnetic resonance (EPR) spectroscopy was used to determine the ability of these C(60)-polymer complexes to scavenge superoxide. Their cytotoxicity was evaluated in three different cell lines. C(60)-POx and C(60)-PVP complexes exhibited similar physicochemical properties and antioxidant activities. C(60)-poly(2-ethyl-2-oxazoline) (PEtOx) complex, but not C(60)-PVP complex, were efficiently taken up by CATH.a neurons and attenuated the increase in intra-neuronal superoxide induced by angiotensin II (Ang II) stimulation. These results show that C(60)-POx complexes are non-toxic, neuronal cell permeable, superoxide scavenging antioxidants that might be promising candidates for the treatment of brain-related diseases associated with increased levels of superoxide.

Studies on poly-3-hydroxyoctanoate biosynthesis by a consortium of microorganisms

Directory of Open Access Journals (Sweden)

Eremia Mihaela Carmen

2016-06-01

Full Text Available Polyhydroxyalcanoates (PHAs are specifically produced by a wide variety of bacteria, as an intracellular energy reserve in the form of homo- and copolymers of [R]-β-hydroxyalkanoic acids, depending on the C source used for microorganism growth, when the cells are grown under stressing conditions. In this paper we present microbiological accumulation of poly-3-hydroxyoctanoate (PHO by using a consortium of bacterial strains, Pseudomonas putida and Bacillus subtilis, in a rate of 3:1, grown on a fermentation medium based on sodium octanoate as the sole carbon source. The experiments performed in the above mentioned conditions led to the following results: from 18.70 g sodium octanoate (7.72 g/L in the fermentation medium used up during the bioprocess, 3.93-3.96 g/L dry bacterial biomass and 1.834 - 1.884 g/L PHA, containing 85.83 - 86.8% PHO, were obtained.

Multifunctional triblock co-polymer mP3/4HB-b-PEG-b-lPEI for efficient intracellular siRNA delivery and gene silencing.

Science.gov (United States)

Zhou, Li; Chen, Zhifei; Wang, Feifei; Yang, Xiuqun; Zhang, Biliang

2013-04-01

A non-viral siRNA carrier composed of mono-methoxy-poly (3-hydroxybutyrate-co-4-hydroxybutyrate)-block-polyethylene glycol-block-linear polyethyleneimine (mP3/4HB-b-PEG-b-lPEI) was synthesized using 1800 Da linear polyethyleneimine and evaluated for siRNA delivery. Our study demonstrated that siRNA could be efficiently combined with mP3/4HB-b-PEG-b-lPEI (mAG) co-polymer and was protected from nuclease degradation. The combined siRNA were released from the complexes easily under heparin competition. The particle size of the mAG/siRNA complexes was 158 nm, with a ζ-potential of around 28 mV. Atomic force microscopy images displayed spherical and homogeneously distributed complexes. The mAG block co-polymer displayed low cytotoxicity and efficient cellular uptake of Cy3-siRNA in A549 cells by flow cytometry and confocal microscopy. In vitro transfection efficiency of the block co-polymer was assessed using siRNA against luciferase in cultured A549-Luc, HeLa-Luc, HLF-Luc, A375-Luc and MCF-7-Luc cells. A higher transfection efficiency and lower cytotoxicity was obtained by mAG block co-polymer in five cell lines. Furthermore, a remarkable improvement in luciferase gene silencing efficiency of the mAG complex (up to 90-95%) over that of Lipofectamine™ 2000 (70-82%) was observed in HLF-Luc and A375-Luc cells. Additionally, a mAG/p65-siRNA complex also showed a better capability than Lipofectamine™ 2000/p65-siRNA complex to drastically reduce the p65 mRNA level down to 10-16% in HeLa, U251 and HUVEC cells at an N/P ratio of 70. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

Effect of dietary poly-β-hydroxybutyrate (PHB) on growth performance, intestinal health status and body composition of Pacific white shrimp Litopenaeus vannamei (Boone, 1931).

Science.gov (United States)

Duan, Yafei; Zhang, Yue; Dong, Hongbiao; Zheng, Xiaoting; Wang, Yun; Li, Hua; Liu, Qingsong; Zhang, Jiasong

2017-01-01

In the present study, the effect of dietary supplementation of poly-β-hydroxybutyrate (PHB) on the growth performance, intestinal digestive and immune function, intestinal short-chain fatty acids (SCFA) content and body composition of Pacific white shrimp Litopenaeus vannamei (Boone, 1931) was evaluated. The shrimp was fed for 35 days with four different diets: 0%, 1%, 3% and 5% PHB supplemented feed. The results indicated that supplementation of PHB significantly increased the growth performance of the shrimp, and the feed conversion rate (FCR) in 3%PHB treatment group was significantly lower than the control (P PHB treatment groups were all significantly higher than that of the control (P PHB treatment (P > 0.05). The activities of intestinal immune enzymes such as total antioxidant capacity (T-AOC) and inducible nitric oxide synthase (iNOS) was significantly induced by 3%PHB treatment (P PHB treatment and nitric oxide (NO) content was significantly induced in three PHB treatments. Meanwhile, PHB induced significantly the expression level of intestinal heat shock protein 70 (HSP70), Toll and immune deficiency (Imd) gene. HE staining showed that PHB induced the intestinal health status of L. vannamei. Intestinal SCFA content analysis revealed that the content of propionic and butyric acid of 3%PHB treatment were significantly higher than that of the control (P PHB treatments, and the crude lipid in 1% and 5%PHB treatments were all significantly higher than the control (P PHB could improve the growth performance, modulated intestinal digestive and immune function, increased intestinal SCFA content and body composition in L. vannamei, and the optimum dietary PHB requirement by L. vannamei was estimated at 3% (w/w) diet. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Opportunity for High-Performance Biomaterials from Methane

Directory of Open Access Journals (Sweden)

Peter James Strong

2016-02-01

Full Text Available Polyhydroxyalkanoate (PHA biopolymers are widely recognised as outstanding candidates to replace conventional petroleum-derived polymers. Their mechanical properties are good and can be tailored through copolymer composition, they are biodegradable, and unlike many alternatives, they do not rely on oil-based feedstocks. Further, they are the only commodity polymer that can be synthesised intracellularly, ensuring stereoregularity and high molecular weight. However, despite offering enormous potential for many years, they are still not making a significant impact. This is broadly because commercial uptake has been limited by variable performance (inconsistent polymer properties and high production costs of the raw polymer. Additionally, the main type of PHA produced naturally is poly-3-hydroxybutyrate (PHB, which has limited scope due to its brittle nature and low thermal stability, as well as its tendency to embrittle over time. Production cost is strongly impacted by the type of the feedstock used. In this article we consider: the production of PHAs from methanotrophs using methane as a cost-effective substrate; the use of mixed cultures, as opposed to pure strains; and strategies to generate a poly(3-hydroxybutyrate-co-3-hydroxyvalerate copolymer (PHBV, which has more desirable qualities such as toughness and elasticity.

Synthesis of zinc sulfide nanoparticles and their incorporation into poly(hydroxybutyrate) matrix in the formation of a novel nanocomposite

Science.gov (United States)

Riaz, Shahina; Raza, Zulfiqar Ali; Majeed, Muhammad Irfan; Jan, Tariq

2018-05-01

In the present study, zinc sulfide (ZnS) nanoparticles (NPs) were successfully synthesized through a modified chemical precipitation protocol and then mediated into poly(hydroxybutyrate) (PHB) matrix to get ZnS/PHB nanocomposite. Mean diameter and zeta potential of ZnS NPs, as determined using dynamic light scattering technique (DLS), were observed to be 53 nm and ‑89 mV, respectively. The structural investigations performed using x-ray diffraction (XRD) technique depicted the phase purity of ZnS NPs exhibiting cubic crystal structure. Fourier transform infrared (FTIR) spectroscopic analysis was conducted to identify the presence or absence of bonding vibrational modes on the surface of synthesized single phase ZnS NPs. The FTIR analysis confirmed the metal to sulphur bond formation by showing the characteristic band at 1123 cm‑1. The UV–vis absorption spectra of ZnS NPs confirmed the synthesis of particles in nanoscale regime showing a λ max of 302 nm. These NPs were then successfully incorporated into PHB matrix to synthesize ZnS/PHB nanocomposite. The synthesis of nanocomposite was confirmed by EDX analysis. The chemical bonding and structural properties of ZnS/PHB nanocomposite were determined by FTIR and XRD analysis, respectively. The FTIR analysis confirmed the synthesis of ZnS/PHB nanocomposite. Moreover, XRD analysis showed that structure of nanocomposite was completely controlled by ZnS NPs as pure PHB exhibited orthorhombic crystal structure while the nanocomposite demonstrated cubic crystal structure of ZnS. Thermal properties of nanocomposite were studied through thermogravimetric analysis revealing that the incorporation of ZnS NPs into PHB matrix lead to enhance heat resistance properties of PHB.

Environmental analysis of plastic production processes: comparing petroleum-based polypropylene and polyethylene with biologically-based poly-beta-hydroxybutyric acid using life cycle analysis.

Science.gov (United States)

Harding, K G; Dennis, J S; von Blottnitz, H; Harrison, S T L

2007-05-31

Polymers based on olefins have wide commercial applicability. However, they are made from non-renewable resources and are characterised by difficulty in disposal where recycle and re-use is not feasible. Poly-beta-hydroxybutyric acid (PHB) provides one example of a polymer made from renewable resources. Before motivating its widespread use, the advantages of a renewable polymer must be weighed against the environmental aspects of its production. Previous studies relating the environmental impacts of petroleum-based and bio-plastics have centred on the impact categories of global warming and fossil fuel depletion. Cradle-to-grave studies report equivalent or reduced global warming impacts, in comparison to equivalent polyolefin processes. This stems from a perceived CO(2) neutral status of the renewable resource. Indeed, no previous work has reported the results of a life cycle assessment (LCA) giving the environmental impacts in all major categories. This study investigates a cradle-to-gate LCA of PHB production taking into account net CO(2) generation and all major impact categories. It compares the findings with similar studies of polypropylene (PP) and polyethylene (PE). It is found that, in all of the life cycle categories, PHB is superior to PP. Energy requirements are slightly lower than previously observed and significantly lower than those for polyolefin production. PE impacts are lower than PHB values in acidification and eutrophication.

A life cycle assessment of poly-hydroxybutyrate extraction from microbial biomass using dimethyl carbonate

DEFF Research Database (Denmark)

Righi, Serena; Baioli, Filippo; Samorì, Chiara

2017-01-01

Poly-hydroxyalkanoates are an example of biodegradable and biocompatible polymers, produced from renewable raw materials. With respect to other bioplastics the market share of poly-hydroxyalkanoates is still limited because of their commercial costs. To develop more cost-effective processes, a mu...... by evaporation appears to be the most promising in terms of environmental sustainability performance....

Production of (R)-3-hydroxybutyric acid by fermentation and bioconversion processes with Azohydromonas lata.

Science.gov (United States)

Ugwu, Charles U; Tokiwa, Yutaka; Ichiba, Toshio

2011-06-01

Feasibility of producing (R)-3-hydroxybutyric acid ((R)-3-HB) using wild type Azohydromonas lata and its mutants (derived by UV mutation) was investigated. A. lata mutant (M5) produced 780 m g/l in the culture broth when sucrose was used as the carbon source. M5 was further studied in terms of its specificity with various bioconversion substrates for production of (R)-3-HB. (R)-3-HB concentration produced in the culture broth by M5 mutant was 2.7-fold higher than that of the wild type strain when sucrose (3% w/v) and (R,S)-1,3-butanediol (3% v/v) were used as carbon source and bioconversion substrate, respectively. Bioconversion of resting cells (M5) with glucose (1% v/w), ethylacetoacetate (2% v/v), and (R,S)-1,3-butanediol (3% v/v), resulted in (R)-3-HB concentrations of 6.5 g/l, 7.3g/l and 8.7 g/l, respectively. Copyright © 2011 Elsevier Ltd. All rights reserved.

Non-isothermal cold crystallization kinetics of poly(3-hydoxybutyrate) filled with zinc oxide

Energy Technology Data Exchange (ETDEWEB)

Ries, Andreas, E-mail: ries750@yahoo.com.br [Electrical Engineering Department, Federal University of Paraíba, João Pessoa, PB 58051-900 (Brazil); Canedo, Eduardo L. [Materials Engineering Department, Federal University of Campina Grande, Campina Grande, PB 58429-900 (Brazil); Souto, Cícero R. [Electrical Engineering Department, Federal University of Paraíba, João Pessoa, PB 58051-900 (Brazil); Wellen, Renate M.R. [Materials Engineering Department, Federal University of Paraíba, João Pessoa, PB 58051-900 (Brazil)

2016-08-10

Highlights: • Non-isothermal cold crystallization kinetics of PHB filled with ZnO is presented. • Pseudo-Avrami model is best for describing an individual crystallization condition. • Mo model is allows to judge the kinetics of a condition untested in this work. • ZnO affects the kinetics irregularly. - Abstract: The non-isothermal cold crystallization kinetics of poly(3-hydroxybutyrate) (PHB) and PHB-ZnO composites, with ZnO content of 1%, 5% and 10% per weight, was investigated at different heating rates (5, 7.5, 10, 15, 20 and 30 °C/min) using differential scanning calorimetry. Both, Kissinger and Friedman activation energies predict correctly the slowest and fastest crystallizing composition. It was further found, that ZnO can neither be classified as a crystallization accelerator, nor as a crystallization inhibitor; its action is strongly concentration dependent. The empirical Pseudo-Avrami model has the best overall capability for fitting the experimental kinetic data. However, since the Pseudo-Avrami exponent was found to vary irregularly with heating rate and filler content, this model should not be applied for kinetic predictions of an arbitrary composition or an untested heating rate. In such cases, Mo's model should be used.

Poly-β-hydroxybutyrate (PHB) accumulating Bacillus spp. improve the survival, growth and robustness of Penaeus monodon (Fabricius, 1798) postlarvae.

Science.gov (United States)

Laranja, Joseph Leopoldo Q; Ludevese-Pascual, Gladys L; Amar, Edgar C; Sorgeloos, Patrick; Bossier, Peter; De Schryver, Peter

2014-10-10

Low larval survival resulting from suboptimal culture conditions and luminous vibriosis poses a major problem for the larviculture of penaeid shrimp. In this study, a poly-β-hydroxybutyrate (PHB) accumulating mixed bacterial culture (mBC; 48.5% PHB on cell dry weight) and two PHB accumulating bacterial isolates, Bacillus sp. JL47 (54.7% PHB on cell dry weight) and Bacillus sp. JL1 (45.5% PHB on cell dry weight), were obtained from a Philippine shrimp culture pond and investigated for their capacity to improve growth, survival and robustness of Penaeus monodon postlarvae (PL). Shrimp PL1 and shrimp PL30 were provided with the PHB containing bacterial cultures in the feed for 30 days followed by, respectively, a challenge with pathogenic Vibrio campbellii and exposure to a lethal dose of ammonia. Prior to the pathogenic challenge or ammonia stress, growth and survival were higher for shrimp receiving the PHB accumulating bacteria as compared to shrimp receiving diets without bacterial additions. After exposure to the pathogenic challenge the shrimp fed PHB accumulating bacteria showed a higher survival as compared to non-treated shrimp, suggesting an increase in robustness for the shrimp. Similar effects were observed when shrimp PL30 were provided with the PHB accumulating bacterial cultures during a challenge with pathogenic V. campbellii through the water. The survival of shrimp exposed to lethal ammonia stress showed no significant difference between PHB accumulating bacteria-fed shrimp and non-PHB treated shrimp. The data illustrate that bacilli capable of accumulating PHB can provide beneficial effects to P. monodon post-larvae during culture in terms of growth performance, survival and resistance against pathogenic infection and ammonia stress. Further investigations are required to verify the PHB effect of the bacterial cultures on the shrimp. Copyright © 2014 Elsevier B.V. All rights reserved.

Evaluation of a fully absorbable poly-4-hydroxybutyrate/absorbable barrier composite mesh in a porcine model of ventral hernia repair.

Science.gov (United States)

Scott, Jeffrey R; Deeken, Corey R; Martindale, Robert G; Rosen, Michael J

2016-09-01

The objective of this study was to evaluate the mechanical and histological properties of a fully absorbable poly-4-hydroxybutyrate/absorbable barrier composite mesh (Phasix™ ST) compared to partially absorbable (Ventralight™ ST), fully absorbable (Phasix™), and biologically derived (Strattice™) meshes in a porcine model of ventral hernia repair. Bilateral abdominal surgical defects were created in twenty-four Yucatan pigs, repaired with intraperitoneal (Phasix™ ST, Ventralight™ ST) or retromuscular (Phasix™, Strattice™) mesh, and evaluated at 12 and 24 weeks (n = 6 mesh/group/time point). Prior to implantation, Strattice™ demonstrated significantly higher (p weeks, mesh/repair strength was significantly greater than NAW (p weeks (p > 0.05). Phasix™ mesh/repair strength was significantly greater than Strattice™ (p weeks, and Ventralight™ ST mesh/repair strength was significantly greater than Phasix™ ST mesh (p weeks. At 12 and 24 weeks, Phasix™ ST and Ventralight™ ST were associated with mild inflammation and minimal-mild fibrosis/neovascularization, with no significant differences between groups. At both time points, Phasix™ was associated with minimal-mild inflammation/fibrosis and mild neovascularization. Strattice™ was associated with minimal inflammation/fibrosis, with minimal neovascularization at 12 weeks, which increased to mild by 24 weeks. Strattice™ exhibited significantly less neovascularization than Phasix™ at 12 weeks and significantly greater inflammation at 24 weeks due to remodeling. Phasix™ ST demonstrated mechanical and histological properties comparable to partially absorbable (Ventralight™ ST) and fully resorbable (Phasix™) meshes at 12 and 24 weeks in this model. Data also suggest that fully absorbable meshes with longer-term resorption profiles may provide improved mechanical and histological properties compared to biologically derived scaffolds.

3-Hydroxybutyrate regulates energy metabolism and induces BDNF expression in cerebral cortical neurons.

Science.gov (United States)

Marosi, Krisztina; Kim, Sang Woo; Moehl, Keelin; Scheibye-Knudsen, Morten; Cheng, Aiwu; Cutler, Roy; Camandola, Simonetta; Mattson, Mark P

2016-12-01

During fasting and vigorous exercise, a shift of brain cell energy substrate utilization from glucose to the ketone 3-hydroxybutyrate (3OHB) occurs. Studies have shown that 3OHB can protect neurons against excitotoxicity and oxidative stress, but the underlying mechanisms remain unclear. Neurons maintained in the presence of 3OHB exhibited increased oxygen consumption and ATP production, and an elevated NAD + /NADH ratio. We found that 3OHB metabolism increases mitochondrial respiration which drives changes in expression of brain-derived neurotrophic factor (BDNF) in cultured cerebral cortical neurons. The mechanism by which 3OHB induces Bdnf gene expression involves generation of reactive oxygen species, activation of the transcription factor NF-κB, and activity of the histone acetyltransferase p300/EP300. Because BDNF plays important roles in synaptic plasticity and neuronal stress resistance, our findings suggest cellular signaling mechanisms by which 3OHB may mediate adaptive responses of neurons to fasting, exercise, and ketogenic diets. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

A new disease? Persistent isolated beta.hydroxybutyrate ketosis and mild congenital hyperinsulinism

DEFF Research Database (Denmark)

Christesen, Henrik Thybo; Brusgaard, Klaus; Ditkovskaya, L

A new disease? Persistent isolated beta.hydroxybutyrate ketosis and mild congenital hyperinsulinism.......A new disease? Persistent isolated beta.hydroxybutyrate ketosis and mild congenital hyperinsulinism....

Novel Sustainable Composites Based on Poly(hydroxybutyrate-co-hydroxyvalerate) and Seagrass Beach-CAST Fibers: Performance and Degradability in Marine Environments

Science.gov (United States)

Mallegni, Norma; Stefanelli, Eleonora; Rossi, Alessia

2018-01-01

In order to produce sustainable, bio-based and highly biodegradable materials, composites based on poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and fibers of Posidonia oceanica (PO), a dominant Mediterranean seagrass, were produced by simple melt mixing and characterized in terms of thermal stability, morphology and rheological/mechanical properties. In view of their potential application in marine environments, degradation of the developed composites was evaluated under simulated and real marine environmental conditions for 1 year. Using 10 wt % of acetyl tributyl citrate (ATBC) as a plasticizer, smooth processing was achieved for up to 30 wt % of PO fibers, despite the reduction of the melt fluidity observed with increasing fiber loading. The tensile modulus slightly increased (from 2 to 2.4 GPa) while the tensile strength and the elongation decreased (from 23.6 to 21.5 MPa and from 3.2 to 1.9%, respectively) by increasing the PO fiber content from 0 to 30 wt %. Interestingly, the impact resistance of the composites increased with the increasing of the PO content: the Charpy’s impact energy increased from 3.6 (without fiber) to 4.4 kJ/m2 for the composite with 30 wt %. The results of the aerobic biodegradation under simulated marine conditions showed that the presence of PO fibers favored the physical disintegration of the composite increasing the biodegradation rate of the polymeric matrix: after 216 days, the composite with 20 wt % PO fibers showed a biodegradability of about 30% compared to 20% of the composite without fibers. Under real marine conditions, the specimens containing PO fibers showed higher weight losses and deterioration of tensile properties compared to those without fibers. Presumably, biodegradation occurred after colonization of the specimen, and the specimens with 20 wt % PO fibers showed well-developed biofilm consisting of bacteria and fungi on the surface after only 3 months of incubation in marine sediments, unlike the no

Electro-oxidation of chlorophenols on poly(3,4-ethylenedioxythiophene)-poly(styrene sulphonate) composite electrode

International Nuclear Information System (INIS)

Pigani, L.; Musiani, M.; Pirvu, C.; Terzi, F.; Zanardi, C.; Seeber, R.

2007-01-01

The electrochemical behaviour of chlorinated phenols on Pt/poly(3,4-ethylenedioxy)thiophene,LiClO 4 and on Pt/poly(3,4-ethylenedioxy)thiophene,poly(sodium-4-styrenesulphonate) electrodes has been investigated in phosphate buffer solution. Poly(sodium-4-styrenesulphonate) exerts remarkable effect against the electrode fouling induced by oxidation of chlorophenols, allowing us to record the relevant anodic response even after repeated potential cycles. Hypotheses about the role exerted by poly(sodium 4-styrenesulphonate) are made, on the basis of evidences provided by several techniques, such as cyclic voltammetry, electrochemical impedance spectroscopy, electrochemical microgravimetry and atomic force microscopy. Thanks to the fact that different chlorophenols show differences in the voltammetric responses, depending on number and position of the chloro substituents on the aromatic ring, applications of the modified electrode in the analysis of mixtures of chlorinated phenols are possible

Glicólise do poli(3-hidroxibutirato por via enzimática

Directory of Open Access Journals (Sweden)

Everton Luiz de Paul

2014-06-01

Full Text Available Poly(3-hydroxybutyrate, PHB, is a polymer with broad potential applications because of its biodegradability and biocompatibility. However, its high crystallinity is a limiting factor for many applications. To overcome this drawback, one strategy currently employed involves the reduction of the molecular weight of PHB with the concomitant formation of end-functionalized chains, such as those obtained via glycolysis. The glycolysis of PHB can be catalyzed by acid, base, or organometallic compounds. However, to our knowledge, there are no reports regarding PHB glycolysis catalyzed enzymatically. Among the major types of enzymes used in biocatalysis, the lipases stand out because they have the ability to catalyze reactions in both aqueous and organic media. Thus, in this study, we performed the enzymatic glycolysis of PHB using the lipase Amano PS (Pseudomonas cepacia with ethane-1,2-diol (ethylene glycol as the functionalizing agent. The results indicated that the glycolysis was successful and afforded hydroxyl-terminated oligomeric PHB polyols. Nuclear magnetic resonance spectra of the products showed characteristic signals for the terminal hydroxyl groups of the polyols, while thermogravimetric and differential scanning calorimetry analyses confirmed an increase in the thermal stability and a decrease in the crystallinity of the polyols compared with the starting PHB polymer, which were both attributed to the reduction in the molecular weight due to glycolysis.

Avaliação da Biodegradação de Poli-beta-(Hidroxibutirato, Poli-beta-(Hidroxibutirato-co-valerato e Poli-épsilon-(caprolactona em Solo Compostado Evaluation of PHB, PHB-V and PCL biodegradation in composted soil

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Derval S. Rosa

2002-10-01

Full Text Available Na sociedade contemporânea, uma solução ou um equacionamento minimamente satisfatório para os graves problemas ambientais decorrentes da larga utilização de polímeros tem se constituído num dos grandes desafios colocados para a comunidade científica. Na busca por soluções desejáveis, e como alternativa aos polímeros convencionais, os polímeros biodegradáveis têm sido alçados a uma posição de destaque. Dessa maneira, as pesquisas em torno desses compostos também vão adquirindo fundamental importância. Nesse trabalho, apresentamos os resultados da avaliação da biodegradação do poli-beta-(hidroxibutirato (PHB, o poli-beta-(hidroxibutirato-co-valerato (PHB-V e o poli-(épsilon-caprolactona (PCL em solo compostado, utilizando a técnica de biodegradação aeróbia conhecida como Teste de Sturm. As análises térmicas destes polímeros foram realizadas usando calorimetria exploratória diferencial (DSC e a temperatura de fusão e a cristalinidade, para cada polímero, foram determinadas. Os resultados mostraram que o PHB degradou mais rápido que os dois outros polímeros, provavelmente por causa da sua estrutura química que pode ser atacada mais facilmente pelos microorganismos.In contemporary society, a solution or a minimally satisfying approach to the serious environmental problems stemming from the widespread use of polymers has been one of the great challenges the scientific community has had to cope with. The studies around these compounds, including their biodegradability, have acquired fundamental importance. In this work, we present the results from an evaluation of the biodegradation of poly-beta-(hydroxybutyrate (PHB, poly-beta-(hydroxybutyrate-co-valerate (PHB-V and poly-(epsilon-caprolactone (PCL, with products obtained from composting of municipal solid wastes, using the technique of aerobic biodegradation characterization known as Sturm Test. The thermal analysis of these polymers was done using differential

Discovery of the beta-form crystal structure in electrospun nanofibers of bio-based poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] and its implication on properties

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Gong, Liang

Bacterially produced poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (PHBHx) is a new type of bioplastic which not only inherits the excellent biodegradability and biocompatibility of its parent homopolymer, polyhydroxybutyrate (PHB), but also overcomes PHB’s brittleness and stiffness with the incorporation of 3-hydroxyhexanoate (Hx) comonomer units with medium-chain-length (mcl) side chains. The tough and ductile PHBHx, with a much lower crystallinity and melting temperature, is well-suited for many practical applications. Efforts have been made to broaden the application range of PHBHx by introducing the beta-form crystalline structure, where the molecular chains adopt a planar zig-zag conformation. However, it is extremely difficult to produce this beta-form in PHBHx due to its much lower crystallinity and much more flexible molecular chains. In this study, we report an approach using the technique of electrospinning. The strain-induced metastable β-form crystalline structure was successfully introduced in PHBHx by collecting the macroscopically aligned electrospun PHBHx nanofibers across the air gap on a piece of aluminum foil and on the tapered edge of a high-speed rotary disk. The presence of the β-form crystal structure in electrospun fiber mats was confirmed by wide-angle X-ray diffraction (WAXD) and Fourier transform infrared spectroscopy (FTIR), with molecular orientation of the polymer chains along the fiber axis revealed by polarized FTIR. Selected area electron diffraction (SAED) and AFM-IR were utilized to investigate the morphological and structural details of individual PHBHx nanofibers. The results demonstrated a coexistence of the thermodynamically stable α-form crystalline structure, where molecular chains adopt a left-handed 21 helical conformation, and the β-form in single fibers. The molecular orientation level and the relative amounts of the two crystalline polymorphs were found to be highly dependent on fiber collection methods

Glycolysis of poly (3-hydroxybutyrate) catalyzed by an enzyme system

International Nuclear Information System (INIS)

Campos, T.F.; Mano, V.

2010-01-01

In this work we report the studies of PHB glycolysis catalyzed by lipase Amano PS (Pseudomonas cepacia) in the presence of 1,2-ethanediol (ethylene glycol). The reactions were performed in toluene:dichloroethane 3:1 (v/v) at 60 deg C, varying reaction time and concentration of ethylene glycol. PHB and the products of glycolysis (polyols) were characterized by FTIR, 1 H-NMR, and TG. The FTIR spectra of polyols showed no significant change compared to the spectrum of PHB. The 1 H-NMR spectra of the products of glycolysis showed signs of interest between 3 and 4.7 ppm, related to the ethylene glycol protons inserted in the polymer chain. By analyzing the thermograms we observed that the polyols are more thermally stable than PHB. (author)

Intracellular Ca2+ release mediates cationic but not anionic poly(amidoamine) (PAMAM) dendrimer-induced tight junction modulation.

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Avaritt, Brittany R; Swaan, Peter W

2014-09-01

Poly(amidoamine) (PAMAM) dendrimers show great promise for utilization as oral drug delivery vehicles. These polymers are capable of traversing epithelial barriers, and have been shown to translocate by both transcellular and paracellular routes. While many proof-of-concept studies have shown that PAMAM dendrimers improve intestinal transport, little information exists on the mechanisms of paracellular transport, specifically dendrimer-induced tight junction modulation. Using anionic G3.5 and cationic G4 PAMAM dendrimers with known absorption enhancers, we investigated tight junction modulation in Caco-2 monolayers by visualization and mannitol permeability and compared dendrimer-mediated tight junction modulation to that of established permeation enhancers. [(14)C]-Mannitol permeability in the presence and absence of phospholipase C-dependent signaling pathway inhibitors was also examined and indicated that this pathway may mediate dendrimer-induced changes in permeability. Differences between G3.5 and G4 in tight junction protein staining and permeability with inhibitors were evident, suggesting divergent mechanisms were responsible for tight junction modulation. These dissimilarities are further intimated by the intracellular calcium release caused by G4 but not G3.5. Based on our results, it is apparent that the underlying mechanisms of dendrimer permeability are complex, and the complexities are likely a result of the density and sign of the surface charges of PAMAM dendrimers. The results of this study will have implications on the future use of PAMAM dendrimers for oral drug delivery.

Poly-ß-hydroxybutyrate content and dose of the bacterial carrier for Artemia enrichment determine the performance of giant freshwater prawn larvae.

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Thai, Truong Quoc; Wille, Mathieu; Garcia-Gonzalez, Linsey; Sorgeloos, Patrick; Bossier, Peter; De Schryver, Peter

2014-06-01

The beneficial effects of poly-β-hydroxybutyrate (PHB) for aquaculture animals have been shown in several studies. The strategy of applying PHB contained in a bacterial carrier has, however, hardly been considered. The effect of administering PHB-accumulated Alcaligenes eutrophus H16 containing 10 or 80 % PHB on dry weight, named A10 and A80, respectively, through the live feed Artemia was investigated on the culture performance of larvae of the giant freshwater prawn (Macrobrachium rosenbergii). Feeding larvae with Artemia nauplii enriched in a medium containing 100 and 1,000 mg L(-1) A80 significantly increased the survival with about 15 % and the development of the larvae with a larval stage index of about 1 as compared to feeding non-enriched Artemia. The survival of the larvae also significantly increased with about 35 % in case of a challenge with Vibrio harveyi. The efficiency of these treatments was equal to a control treatment of Artemia enriched in an 800 mg L(-1) PHB powder suspension, while Artemia enriched in 10 mg L(-1) A80, 100 mg L(-1) A10, and 1,000 mg L(-1) A10 did not bring similar effects. From our results, it can be concluded that PHB supplemented in a bacterial carrier (i.e., amorphous PHB) can increase the larviculture efficiency of giant freshwater prawn similar to supplementation of PHB in powdered form (i.e., crystalline PHB). When the level of PHB in the bacterial carrier is high, similar beneficial effects can be achieved as crystalline PHB, but at a lower live food enrichment concentration expressed on PHB basis.

Effects of β-hydroxybutyrate and isoproterenol on lipolysis in isolated adipocytes from periparturient dairy cows and cows with clinical ketosis.

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van der Drift, S G A; Everts, R R; Houweling, M; van Leengoed, L A M G; Stegeman, J A; Tielens, A G M; Jorritsma, R

2013-06-01

An in vitro model was used to investigate effects of β-hydroxybutyrate and isoproterenol (β-adrenergic receptor agonist) on lipolysis in isolated adipocytes from late pregnant and recently calved dairy cows (n=5) and cows with clinical ketosis (n=3). Incubation with 3.0 mmol/L β-hydroxybutyrate reduced lipolysis in isolated adipocytes. This inhibitory effect was lower in the first lactation week (47%±16%) compared with late pregnancy (71%±6.5%). Incubation with 0.3 μmol/L isoproterenol stimulated lipolysis in isolated adipocytes from periparturient dairy cows. Basal lipolysis resulted in non-esterified fatty acid to glycerol ratios in the incubation media of 2.0±0.23 in prepartum samples, 2.1±0.23 in the first lactation week and 2.2±0.09 in cows with clinical ketosis. β-Hydroxybutyrate reduced lipolysis by 45%±9.6% in isolated adipocytes from cows with clinical ketosis, indicating that impaired feedback of β-hydroxybutyrate may not play a role in the disease etiology. Copyright © 2012 Elsevier Ltd. All rights reserved.

Fasting mitigates immediate hypersensitivity: a pivotal role of endogenous D-beta-hydroxybutyrate.

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Nakamura, Shigeru; Hisamura, Ryuji; Shimoda, Sachiko; Shibuya, Izumi; Tsubota, Kazuo

2014-01-01

Fasting is a rigorous type of dietary restriction that is associate with a number of health benefits. During fasting, ketone bodies significantly increase in blood and become major body fuels, thereby sparing glucose. In the present study, we investigated effects of fasting on hypersensitivity. In addition, we also investigated the possible role of D-beta-hydroxybutyrate provoked by fasting in the attenuation of immediate hypersensitivity by fasting. Effects of fasting on systemic anaphylaxis were examined using rat model of toluene 2, 4-diisocyanate induced nasal allergy. In addition to food restriction, a ketogenic high-fat and low-carbohydrate diet that accelerates fatty acid oxidation and systemic instillation of D-beta-hydroxybutyrate were employed to elevate internal D-beta-hydroxybutyrate concentration. We assessed relationship between degranulation of rat peritoneal mast cells and internal D-beta-hydroxybutyrate concentration in each treatment. Changes in [Ca(2+)]i responses to compound 48/80 were analyzed in fura 2-loaded rat peritoneal mast cells derived from the ketogenic diet and fasting. Immediate hypersensitivity reaction was significantly suppressed by fasting. A significant reduction in mast cells degranulation, induced by mast cell activator compound 48/80, was observed in rat peritoneal mast cells delivered from the 24 hours fasting treatment. In addition, mast cells delivered from a ketogenic diet and D-beta-hydroxybutyrate infusion treatment also had reduced mast cell degranulation and systemic D-beta-hydroxybutyrate concentrations were elevated to similar extent as the fasting state. The peak increase in [Ca(2+)]i was significantly lower in the ketogenic diet and fasting group than that in the control diet group. The results of the present study demonstrates that fasting suppress hypersensitivity reaction, and indicate that increased level of D-beta-hydroxybutyrate by fasting plays an important role, via the stabilization of mast cells, in

Intracellular ion monitoring using a gold-core polymer-shell nanosensor architecture

Energy Technology Data Exchange (ETDEWEB)

Stanca, S E; Cranfield, C G; Biskup, C [Biomolecular Photonics Group, University Hospital Jena, Teichgraben 8, 07743 Jena (Germany); Nietzsche, S [Centre for Electron Microscopy, University Hospital Jena, Ziegel-muehlenweg 1, 07743 Jena (Germany); Fritzsche, W, E-mail: sarmiza.stanca@mti.uni-jena.de, E-mail: charles.cranfield@mti.uni-jena.de, E-mail: christoph.biskup@mti.uni-jena.de [Institute of Photonic Technology, Albert-Einstein-Strasse 9, 07745 Jena (Germany)

2010-02-05

In this study, we describe the design of new ratiometric fluorescent nanosensors, whose architecture is based on a gold core surrounded by a poly(vinyl alcohol)-polyacetal shell. To the gold core, indicator dyes and reference dyes are attached via a cysteine linker. This nanosensor architecture is flexible with regards to the number and types of fluorophore linkages possible. The robust poly(vinyl alcohol)-polyacetal shell protects the fluorophores linked to the core from non-specific interactions with intracellular proteins. The nanosensors developed in this way are biocompatible and can be easily incorporated into mammalian cells without the use of transfection agents. Here, we show the application of these nanosensors for intracellular pH and sodium ion measurements.

Novel Sustainable Composites Based on Poly(hydroxybutyrate-co-hydroxyvalerate and Seagrass Beach-CAST Fibers: Performance and Degradability in Marine Environments

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Maurizia Seggiani

2018-05-01

Full Text Available In order to produce sustainable, bio-based and highly biodegradable materials, composites based on poly(hydroxybutyrate-co-hydroxyvalerate (PHBV and fibers of Posidonia oceanica (PO, a dominant Mediterranean seagrass, were produced by simple melt mixing and characterized in terms of thermal stability, morphology and rheological/mechanical properties. In view of their potential application in marine environments, degradation of the developed composites was evaluated under simulated and real marine environmental conditions for 1 year. Using 10 wt % of acetyl tributyl citrate (ATBC as a plasticizer, smooth processing was achieved for up to 30 wt % of PO fibers, despite the reduction of the melt fluidity observed with increasing fiber loading. The tensile modulus slightly increased (from 2 to 2.4 GPa while the tensile strength and the elongation decreased (from 23.6 to 21.5 MPa and from 3.2 to 1.9%, respectively by increasing the PO fiber content from 0 to 30 wt %. Interestingly, the impact resistance of the composites increased with the increasing of the PO content: the Charpy’s impact energy increased from 3.6 (without fiber to 4.4 kJ/m2 for the composite with 30 wt %. The results of the aerobic biodegradation under simulated marine conditions showed that the presence of PO fibers favored the physical disintegration of the composite increasing the biodegradation rate of the polymeric matrix: after 216 days, the composite with 20 wt % PO fibers showed a biodegradability of about 30% compared to 20% of the composite without fibers. Under real marine conditions, the specimens containing PO fibers showed higher weight losses and deterioration of tensile properties compared to those without fibers. Presumably, biodegradation occurred after colonization of the specimen, and the specimens with 20 wt % PO fibers showed well-developed biofilm consisting of bacteria and fungi on the surface after only 3 months of incubation in marine sediments, unlike the

A life-cycle assessment of poly-hydroxybutyrate extraction from microbial biomass using dimethylcarbonate

DEFF Research Database (Denmark)

Righi, Serena; Baioli, Filippo; Samorì, Chiara

2016-01-01

Plastic materials have wide commercial applicability. However, they are made from non-renewable resources and are characterised by resistance to degradation. Poly-hydroxyalkanoates (PHAs) provides one example of a polymer biodegradable, biocompatible and produced from renewable raw materials...

Viability of HEK 293 cells on poly-β-hydroxybutyrate (PHB) biosynthesized from a mutant Azotobacter vinelandii strain. Cast film and electrospun scaffolds.

Science.gov (United States)

Romo-Uribe, Angel; Meneses-Acosta, Angelica; Domínguez-Díaz, Maraolina

2017-12-01

Sterilization, cytotoxicity and cell viability are essential properties defining a material for medical applications and these characteristics were investigated for poly(β-hydroxybutyrate) (PHB) of 230kDa obtained by bacterial synthesis from a mutant strain of Azotobacter vinelandii. Cell viability was investigated for two types of PHB scaffolds, solution cast films and non-woven electrospun fibrous membranes, and the efficiency was compared against a culture dish. The biosynthesized PHB was sterilized by ultraviolet radiation and autoclave, it was found that the thermal properties and intrinsic viscosity remained unchanged indicating that the sterilization methods did not degrade the polymer. Sterilized scaffolds were then seeded with human embryonic kidney 293 (HEK 293) cells to evaluate the cytotoxic response. The cell viability of these cells was evaluated for up to six days, and the results showed that the cell morphology was normal, with no cytotoxic effects. The films and electrospun membranes exhibited over 95% cell viability whereas the viability in culture dishes reached only ca. 90%. The electrospun membrane, however, exhibited significantly higher cell density than the cast film suggesting that the fibrous morphology enables better nutrients transfer. The results indicate that the biosynthesized PHB stands UV and autoclave sterilization methods, it is biocompatible and non-toxic for cell growth of human cell lines. Furthermore, cell culture for up to 18 days showed that 62% and 90% of mass was lost for the film and fibrous electrospun scaffold, respectively. This is a favorable outcome for use in tissue engineering where material degradation, as tissue regenerates, is desirable. Copyright © 2017 Elsevier B.V. All rights reserved.

Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems: distinct tools for intracellular delivery of cDNA and siRNA.

Science.gov (United States)

McLaggan, Debra; Adjimatera, Noppadon; Sepcić, Kristina; Jaspars, Marcel; MacEwan, David J; Blagbrough, Ian S; Scott, Roderick H

2006-01-16

Haplosclerid marine sponges produce pore forming polyalkylpyridinium salts (poly-APS), which can be used to deliver macromolecules into cells. The aim of this study was to investigate the delivery of DNA, siRNA and lucifer yellow into cells mediated by poly-APS and its potential mechanisms as compared with other lipofection systems (lipofectamine and N4,N9-dioleoylspermine (LipoGen)). DNA condensation was evaluated and HEK 293 and HtTA HeLa cells were used to investigate pore formation and intracellular delivery of cDNA, siRNA and lucifer yellow. Poly-APS and LipoGen were both found to be highly efficient DNA condensing agents. Fura-2 calcium imaging was used to measure calcium transients indicative of cell membrane pore forming activity. Calcium transients were evoked by poly-APS but not LipoGen and lipofectamine. The increases in intracellular calcium produced by poly-APS showed temperature sensitivity with greater responses being observed at 12 degrees C compared to 21 degrees C. Similarly, delivery of lucifer yellow into cells with poly-APS was enhanced at lower temperatures. Transfection with cDNA encoding for the expression enhanced green fluorescent protein was also evaluated at 12 degrees C with poly-APS, lipofectamine and LipoGen. Intracellular delivery of siRNA was achieved with knockdown in beta-actin expression when lipofectamine and LipoGen were used as transfection reagents. However, intracellular delivery of siRNA was not achieved with poly-APS. Poly-APS mediated pore formation is critical to its activity as a transfection reagent, but lipofection systems utilise distinct mechanisms to enable delivery of DNA and siRNA into cells.

Pore forming polyalkylpyridinium salts from marine sponges versus synthetic lipofection systems: distinct tools for intracellular delivery of cDNA and siRNA

Directory of Open Access Journals (Sweden)

Blagbrough Ian S

2006-01-01

Full Text Available Abstract Background Haplosclerid marine sponges produce pore forming polyalkylpyridinium salts (poly-APS, which can be used to deliver macromolecules into cells. The aim of this study was to investigate the delivery of DNA, siRNA and lucifer yellow into cells mediated by poly-APS and its potential mechanisms as compared with other lipofection systems (lipofectamine and N4,N9-dioleoylspermine (LipoGen. DNA condensation was evaluated and HEK 293 and HtTA HeLa cells were used to investigate pore formation and intracellular delivery of cDNA, siRNA and lucifer yellow. Results Poly-APS and LipoGen were both found to be highly efficient DNA condensing agents. Fura-2 calcium imaging was used to measure calcium transients indicative of cell membrane pore forming activity. Calcium transients were evoked by poly-APS but not LipoGen and lipofectamine. The increases in intracellular calcium produced by poly-APS showed temperature sensitivity with greater responses being observed at 12°C compared to 21°C. Similarly, delivery of lucifer yellow into cells with poly-APS was enhanced at lower temperatures. Transfection with cDNA encoding for the expression enhanced green fluorescent protein was also evaluated at 12°C with poly-APS, lipofectamine and LipoGen. Intracellular delivery of siRNA was achieved with knockdown in beta-actin expression when lipofectamine and LipoGen were used as transfection reagents. However, intracellular delivery of siRNA was not achieved with poly-APS. Conclusion Poly-APS mediated pore formation is critical to its activity as a transfection reagent, but lipofection systems utilise distinct mechanisms to enable delivery of DNA and siRNA into cells.

Breakfast, plasma glucose and beta-hydroxybutyrate, body mass index and academic performance in children from Extremadura, Spain.

Science.gov (United States)

Torres, M D; Carmona, I; Campillo, C; Pérez, G; Campillo, J E

2007-01-01

Nutritional aspects of breakfast, plasma levels of glucose and beta-hydroxybutyrate, body mass index and academic performance have been studied in urban and rural children (Extremadura, Spain). Representative samples of schoolchildren (3 to 12 years old, random cluster-sampling in schools). Children's mean caloric intake with breakfast was 331 kcal. Rural population ingested more carbohydrates (46,9 +/- 12,3% versus 43,3 +/- 13,2% of the total caloric intake) and fewer lipids (40,5 +/- 11,8% versus 43,9 +/- 12,8% of the total caloric intake) than the urban population. Academic performance was significantly better in the children inhabiting the rural zone than in those of the urban zone. The glycaemia was higher in the urban than in the rural children, and that the contrary was the case for the beta-hydroxybutyrate values. Neither glucose nor beta-hydroxybutyrate levels were correlated with academic performance values. BMI was significantly increased in the urban versus rural children. The present results emphasize the importance of breakfast and life style in the weight and the academic performance of children.

Synthesis of racemic, R- and S-[1-11C]-β-hydroxybutyric acid

International Nuclear Information System (INIS)

Thorell, J.-O.; Stone-Elander, S.; Karolinska Hospital and Inst., Stockholm; Koenig, W.A.; Halldin, C.; Widen, L.

1991-01-01

Racemic, R- and S-β-hydroxybutyric acid were labelled with 11 C in the carboxylic position by a two-step stereospecific synthesis starting with carrier-added [ 11 C]cyanide and R/S, R- or S-propylene oxide. Hydrolysis of the intermediate nitrile with hydrochloric acid gave racemic [1- 11 C]-β-hydroxybutyric acid and R- or S-[1- 11 C]-β-hydroxybutyric acid with an enantiomeric excess of 87-97%. The total synthesis time (including HPLC purification) was 45-50 min from end of trapping. The isolated decay-corrected radiochemical yield was 20-30% based on [ 11 C]cyanide. The radiochemical purity of the products was > 99%]. (author)

A novel autolysis system controlled by magnesium and its application to poly (3-hydroxypropionate) production in engineered Escherichia coli.

Science.gov (United States)

Tamekou Lacmata, Stephen; Yao, Lan; Xian, Mo; Liu, Hui; Kuiate, Jules-Roger; Liu, Huizhou; Feng, Xinjun; Zhao, Guang

2017-09-03

The release of intracellular products, especially polyhydroxyalkanoates, is still a great challenge in industry. To solve this bottleneck, a novel autolysis system strictly controlled with magnesium was constructed and applied to poly(3-hydroxypropionate) production in engineered Escherichia coli. The autolysis system was constructed by inserting the 5'untranslated region (5'UTR) behind promoter PmgtA with lysis genes (S, R, and Rz, from E. coli) overexpressed. The autolysis system functioned well (lysis efficiency of more than 90%) in the P3HP producer with double plasmids containing lysis genes and P3HP biosynthesis genes, whereas the P3HP production was reduced due to plasmid losses. After the autolysis genes and P3HP biosynthesis genes were integrated into one plasmid, the P3HP content of 72.7% (2.4 times of the control) and the plasmid stability of 79.8 ± 3.1% were achieved in strain Q2646 with promoter PmgtA-UTR. However, the strain Q2647 with promoter PmgtA could not accumulate P3HP because of rapid cell lysis. The novel autolysis system activated in Mg 2+ -depleted conditions proves to be feasible for polyhydroxyalkanoates production, which may have great application potential for other intracellular products.

Radiation effects on biodegradable polyesters

International Nuclear Information System (INIS)

Hiroshi Mitomo; Darmawan Darwis; Fumio Yoshii; Keizo Makuuchi

1999-01-01

Poly(3-hydroxybutyrate) [P(3HB)] and its copolymer poly(3-hydroxybutyrate-co-3hydroxyvalerate) [P(3HB-co-3HV)] are microbial biodegradable polyesters produced by many types of bacteria. Poly(butylene succinate) (PBS) and poly(E-caprolactone) (PCL) are also biodegradable synthetic polyesters which have been commercialized. These thermoplastics are expected for wide usage in environmental protection and blocompatible applications. Radiation grafting of hydrophilic monomers onto many polymers, e.g., polyethylene and polypropylene has been studied mainly for biomedical applications. In the present study, radiation-induced graft polymerization of vinyl monomers onto PHB and P(3HB-co-3HV) was carried out and improvement of their properties was studied. Changes in the properties and biodegradability were compared with the degree of grafting. Radiation-induced crosslinking of PBS and PCL which relatively show thermal and irradiation stability was also carried out to improve their thermal stability or processability. Irradiation to PBS and PCL mainly resulted in crosslinking and characterization of these crosslinked polyesters was investigated

Psychiatric comorbidity, psychological distress, and quality of life in gamma-hydroxybutyrate-dependent patients.

Science.gov (United States)

Kamal, Rama M; Dijkstra, Boukje A G; de Weert-van Oene, Gerdien H; van Duren, Josja A M; de Jong, Cornelis A J

2017-01-01

Understanding the psychiatric state and psychological distress level of patients with gamma-hydroxybutyrate dependence is important to develop effective detoxification and relapse management methods. The aim of the current study was to assess the prevalence among gamma-hydroxybutyrate-dependent individuals of psychiatric comorbidity and psychological distress levels and their association with the individuals' pattern of misuse and quality of life. There were 98 patients tested with the Mini International Neuropsychiatric Interview-plus, the Brief Symptom Inventory, the Depression Anxiety Stress scale, and the EuroQoL-5D as a part of the Dutch gamma-hydroxybutyrate detoxification monitor in 7 addiction treatment centers. Participants were selected from those undergoing inpatient gamma-hydroxybutyrate detoxification treatment between March 2011 and September 2012. Males accounted for 68% of the participants and the average age was 28-years-old. A high rate of psychiatric comorbidity (79%) was detected, including anxiety (current 38%, lifetime 40%), mood (13%, 31%), and psychotic disorders (13%, 21%). The level of psychological distress was significantly higher than the standard outpatient reference group, especially in patients with current psychiatric comorbidity (Brief Symptom Inventory Global Severity Index mean 1.61 versus 1.09, p ≤ 0.01). Increased gamma-hydroxybutyrate misuse (higher dose and shorter interval between doses) was associated with the presence of lifetime psychosis, current mood disorders (r pb = 0.23, p = 0.025), and psychoticism as a symptom of psychological distress. Current anxiety, mood disorders and high psychological stress had a negative effect on participants' quality of life. Gamma-hydroxybutyrate dependence is characterized by serious psychiatric comorbidity and psychological distress, both of which are, in turn, associated with increased gamma-hydroxybutyrate use and a lower quality of life. This needs to be considered during

Glycolysis of poly (3-hydroxybutyrate) catalyzed by an enzyme system; Glicolise do poli(3-hidroxibutirato) por via enzimatica

Energy Technology Data Exchange (ETDEWEB)

Campos, T.F.; Mano, V., E-mail: mano@ufsj.edu.b [Universidade Federal de Sao Joao del Rei (UFSJ), MG (Brazil). Dept. de Ciencias Naturais

2010-07-01

In this work we report the studies of PHB glycolysis catalyzed by lipase Amano PS (Pseudomonas cepacia) in the presence of 1,2-ethanediol (ethylene glycol). The reactions were performed in toluene:dichloroethane 3:1 (v/v) at 60 deg C, varying reaction time and concentration of ethylene glycol. PHB and the products of glycolysis (polyols) were characterized by FTIR, {sup 1}H-NMR, and TG. The FTIR spectra of polyols showed no significant change compared to the spectrum of PHB. The {sup 1}H-NMR spectra of the products of glycolysis showed signs of interest between 3 and 4.7 ppm, related to the ethylene glycol protons inserted in the polymer chain. By analyzing the thermograms we observed that the polyols are more thermally stable than PHB. (author)

Packaging related properties of commercially available biopolymers – An overview of the status quo

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V. Jost

2018-05-01

Full Text Available Several commercially available thermoplastic biopolymers were processed in a continuous extrusion line. The molecular weight, crystallinity, and mechanical and permeation properties of the cast films were determined in order to evaluate the status quo of biopolymers currently commercially available. The biopolymers that were evaluated were polylactic acid (PLA, several polyhydroxyalkanoates (PHAs (Poly(3-hydroxybutyrate (PHB, poly(3-hydroxybutyrate-co-4-hydroxybutyrate (PHBHB, poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV, thermoplastic starch (TPS, polybutylene adipate terephthalate (PBAT, polybutylene succinate (PBS, polycaprolactone (PCL and biobased polyethylene (BioPE. Due to its potential for biobased production, thermoplastic polyurethane elastomer (TPU was also analysed. Mechanical analysis showed the PLA and PHA films had high strength and extremely low elongation at break. These were also the materials with the highest molecular weights. Films made of TPU, PCL, TPS, PBAT and BioPE had a significantly lower Young’s modulus and significantly higher elongation at break; these films had comparatively low molecular weights. Permeation measurements showed that PHA films, and particularly PHBV, had the lowest oxygen and water vapour permeability of the biopolymers that were analysed. The biopolymers BioPE, TPS, PCL, TPU and PBAT were highly permeable to oxygen, and had comparatively low molecular weight. The biopolymers TPU, PBS, PBAT, PCL and TPS were highly permeable to water vapour.

Environmental biodegradation of haloarchaea-produced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in activated sludge

DEFF Research Database (Denmark)

Liu, Xiao-Bin; Wu, Linping; Hou, Jing

2016-01-01

-produced PHBHV films, with 10~60 mol% 3-hydroxyvalerate (3HV) composition and different microchemical structures, was studied in nutrition-depleted activated sludge. The changes in mass, molar mass, chemical composition, thermal properties, and surface morphology were monitored. The mass and molar mass of each...

Intracellular organisation of polyhydroxyalkanoate inclusion bodies: a role for small angle neutron scattering?

International Nuclear Information System (INIS)

Foster, L.J.R.; Holden, P.J.; Garvey, C.J.; Russell, R.A.; Stone, D.J.M.

2003-01-01

Full text: Polyhydroxyalkanoates (PHAs) are a diverse family of bacterially produced biopolyesters. Their biodegradability, and in some cases biocompatibility, suggest applications ranging from bioplastics to biomedical implantation devices. Despite extensive interest in their production and potential applications, little is known about their intracellular organisation. Microbial PHAs are synthesised by microorganisms under conditions of nutrient stress and can comprise up to 90% of the dry cell mass. The formation and organisation of these PHA inclusion bodies requires clarification. Such investigations have important implications for the biotechnological production of PHAs in microbes and other organisms, for downstream processing and in vitro precision polymerisation. Morphological and biochemical evidence supports two different models for the intracellular organisation of PHAs. Steinbuchel and coworkers propose a simple model of amorphous PHA enclosed by a single protein membrane consisting of structural proteins (PHAsins) and enzymes responsible for synthesis and degradation. In contrast, Fuller and coworkers have theorised a more complex system of PHA encompassed by a PHAsin bilayer separated by phospholipid. The polymerase and depolymerase enzymes are proposed to be associated with an incomplete inner PHAsin layer. It may be that such models are genera or species specific, since both proposals were derived from research on different species producing different types of PHA. Our initial investigations have focussed on in vivo deuteration of polyhydroxyoctanoate, produced by Pseudomonas oleovorans, both in fermentation on natural and deuterated substrates and during Small Angle Neutron Scattering by whole cells using AUSANS. The nature of the structural questions and our preliminary findings including contrast variation data will be discussed

Nitrate removal performance of Diaphorobacter nitroreducens using biodegradable plastics as the source of reducing power

Energy Technology Data Exchange (ETDEWEB)

Khan, S. T. [Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi 441-8580, Japan and Department of Zoology, College of Science, King Saud University, Riyadh (Saudi Arabia); Nagao, Y. [Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi 441-8580 (Japan); Hiraishi, A., E-mail: hiraishi@ens.tut.ac.jp [Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi 441-8580, Japan and Electronics-Inspired Interdisciplinary Research Institute (EIIRIS), Toyohashi University of Technology, Toyohashi 441-8580 (Japan)

2015-02-27

Strain NA10B{sup T} and other two strains of the denitrifying betaproteobacterium Diaphorobacter nitroreducens were studied for the performance of solid-phase denitrification (SPD) using poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and some other biodegradable plastics as the source of reducing power in wastewater treatment. Sequencing-batch SPD reactors with these organisms and PHBV granules or flakes as the substrate exhibited good nitrate removal performance. Vial tests using cultures from these parent reactors showed higher nitrate removal rates with PHBV granules (ca. 20 mg-NO{sub 3}{sup −}‐N g{sup −1} [dry wt cells] h{sup −1}) than with PHBV pellets and flakes. In continuous-flow SPD reactors using strain NA10B{sup T} and PHBV flakes, nitrate was not detected even at a loading rate of 21 mg-NO{sub 3}{sup −}‐N L{sup −1} h{sup −1}. This corresponded to a nitrate removal rate of 47 mg-NO{sub 3}{sup −}‐N g{sup −1} (dry wt cells) h{sup −1}. In the continuous-flow reactor, the transcription level of the phaZ gene, coding for PHB depolymerase, decreased with time, while that of the nosZ gene, involved in denitrificaiton, was relatively constant. These results suggest that the bioavailability of soluble metabolites as electron donor and carbon sources increases with time in the continuous-flow SPD process, thereby having much higher nitrate removal rates than the process with fresh PHBV as the substrate.

Large electrically induced height and volume changes in poly(3,4- ethylenedioxythiophene) /poly(styrenesulfonate) thin films

NARCIS (Netherlands)

Charrier, D.S.H.; Janssen, R.A.J.; Kemerink, M.

2010-01-01

We demonstrate large, partly reversible height and volume changes of thin films of poly(3,4-ethylenedioxythiophene)/poly(styrenesulfonate) (PEDOT:PSS) on the anode of interdigitating gold electrodes under ambient conditions by applying an electrical bias. The height and volume changes were monitored

Preparation of poly (styrene)-b-poly (acrylic acid)/{gamma}-Fe{sub 2}O{sub 3} composites

Energy Technology Data Exchange (ETDEWEB)

Zhang, L.D. [School of Materials Science and Engineering, Shandong Polytechnic University, Key Laboratory of Processing and Testing Technology of Glass Functional Ceramics of Shandong Province, Daxue Road, Western University Science Park, Jinan 250353 (China); Liu, W.L., E-mail: wlliu@sdu.edu.cn [School of Materials Science and Engineering, Shandong Polytechnic University, Key Laboratory of Processing and Testing Technology of Glass Functional Ceramics of Shandong Province, Daxue Road, Western University Science Park, Jinan 250353 (China); Xiao, C.L.; Yao, J.S.; Fan, Z.P.; Sun, X.L.; Zhang, X.; Wang, L. [School of Materials Science and Engineering, Shandong Polytechnic University, Key Laboratory of Processing and Testing Technology of Glass Functional Ceramics of Shandong Province, Daxue Road, Western University Science Park, Jinan 250353 (China); Wang, X.Q. [State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100 (China)

2011-12-15

The use of a block copolymer, poly (styrene)-b-poly (acrylic acid) (PS-b-PAA) to prepare a magnetic nanocomposite was investigated. Poly (styrene)-poly (t-butyl acrylate) block copolymer, being synthesized by atom transfer radical polymerization, was hydrolyzed with hydrochloric acid for obtaining PS-b-PAA. The obtained PS-b-PAA was then compounded with the modified {gamma}-Fe{sub 2}O{sub 3}, and subsequently the magnetic nanocomposite was achieved. The products were characterized by {sup 1}H NMR, FTIR, gel permeation chromatography, thermogravimetric analysis, transmission electron microscopy and vibrating sample magnetometer. The results showed that the nanocomposites exhibited soft magnetism, with the mean diameter of 100 nm approximately. - Highlights: > Magnetic composites were prepared using {gamma}-Fe{sub 2}O{sub 3} and PS-b-PAA. > PS-b-PAA was synthesized by atom transfer radical polymerization. > The obtained composite exhibited soft magnetism.

A kinetic model for growth and biosynthesis of medium-chain-length poly-(3-hydroxyalkanoates in Pseudomonas putida

Directory of Open Access Journals (Sweden)

M. S. M. Annuar

2008-06-01

Full Text Available A kinetic model is presented giving a mathematical description of batch culture of Pseudomonas putida PGA1 grown using saponified palm kernel oil as carbon source and ammonium as the limiting nutrient. The growth of the micro-organism is well-described using Tessier-type model which takes into account the inhibitory effect of ammonium at high concentrations. The ammonium consumption rate by the cells is related in proportion to the rate of growth. The intracellular production of medium-chain-length poly-(3-hydroxyalkanoates (PHA MCL by P. putida PGA1 cells is reasonably modeled by the modified Luedeking-Piret kinetics, which incorporate a function of product synthesis inhibition (or reduction by ammonium above a threshold level.

ACCUMULATION OF POLYHYDROXYALKANOIC ACIDS BY AZOTOBACTER CHROOCOCCUM MAL-201 FROM ORGANIC WASTE

Directory of Open Access Journals (Sweden)

Soma Pal Saha

2013-08-01

Full Text Available Azotobacter chroococcum MAL-201 (MTCC 3853, a free-living nitrogen-fixing bacterium accumulated intracellular poly(3-hydroxybutyric acid [P(3HB] accounting 69% of cell dry weight (CDW when grown in nitrogrn-free Stockdale medium containing 2% (w/v glucose. It also produced copolymer of poly(3-hydroxybutyrate co-3-hydroxyvalerate [P(3HB-co-3HV] using glucose as primary carbon source and valerate cas cosubstrate. To make the polymer production cost effective four types of waste material of different origin were tested for growth and polymer production. Stockdale medium supplemented with 1% (w/v waste materials failed to yield good growth and polymer accumulation. Two–step cultivation was adopted for better growth and enhanced polymer accumulation. The candy factory waste was most suitable for synthesis of P(3HB accounting 17.8 and 40.58% using single and two-step cultivation conditions respectively. Wastes of domestic and poultry origin produced P(3HB-co-3HV with 3HV content 28.8 and 21.5 mol% respectively in two-step cultivation. Increase concentration of these wastes resulted in further upliftment of 3HV content of polymer with reduced growth and polymer accumulation. However, at optimum incubation the strain MAL-201 cells accumulated P(3HB 48.5% of CDW (at 40h from candy factory waste and P(3HB-co-3HV 24.75 % of CDW with 3HV 34.65 mol % from domestic waste. Intrinsic viscosity, molecular weight and thermal degradation of the polymers accumulated in the cells grown in glucose, glucose with valerate and glucose with waste were compared.

Radiation-induced G/sub 2/-arrest is reduced by inhibitors of poly(adenosine diphosphoribose) synthetase

International Nuclear Information System (INIS)

Rowley, R.

1985-01-01

Experiments are in progress to test whether poly(adenosine diphosphoribose) synthesis is required for the induction of G/sub 2/-arrest in growing mammalian cells following X-irradiation. A variety of poly(ADPR) synthetase inhibitors have been tested to determine: 1) whether addition of an inhibitor to X-irradiated CHO cells reduces G/sub 2/-arrest; 2) whether compounds structurally similar to poly-(ADPR) synthetase inhibitors but inactive against this enzyme affect radiation-induced G/sub 2/-arrest and 3) whether the concentration dependence for poly(ADPR) synthetase inhibition matches that for G/sub 2/-arrest reduction. G/sub 2/-arrest was measured in X-irradiated (1.5 Gy) CHO cells using the mitotic cell selection technique. Poly(ADPR) synthetase activity was measured in permeabilized cells by /sup 3/H-NAD incorporation. The synthetase inhibitors used were 3-aminobenzamide, benzamide, nicotinamide, 4-acetyl pyridine, caffeine and theophylline. The inactive compounds used were 3-aminobenzoic acid, benzoic acid, nicotinic acid, adenine, adenosine and 3'-deoxyadenosine. Inhibitors of poly(ADPR) synthetase reduced G/sub 2/-arrest while related compounds which produced no enzyme inhibition did not. The concentration dependencies for G/sub 2/-arrest reduction and enzyme inhibition were similar only for methyl xanthines. Further analysis awaits the determination of intracellular drug concentrations

Localization of poly(3-hydroxybutyrate) (PHB) granule-associated proteins during PHB granule formation and identification of two new phasins, PhaP6 and PhaP7, in Ralstonia eutropha H16.

Science.gov (United States)

Pfeiffer, Daniel; Jendrossek, Dieter

2012-11-01

Poly(3-hydroxybutyrate) (PHB) granules are covered by a surface layer consisting of mainly phasins and other PHB granule-associated proteins (PGAPs). Phasins are small amphiphilic proteins that determine the number and size of accumulated PHB granules. Five phasin proteins (PhaP1 to PhaP5) are known for Ralstonia eutropha. In this study, we identified three additional potential phasin genes (H16_B1988, H16_B2296, and H16_B2326) by inspection of the R. eutropha genome for sequences with "phasin 2 motifs." To determine whether the corresponding proteins represent true PGAPs, fusions with eYFP (enhanced yellow fluorescent protein) were constructed. Similar fusions of eYFP with PhaP1 to PhaP5 as well as fusions with PHB synthase (PhaC1), an inactive PhaC1 variant (PhaC1-C319A), and PhaC2 were also made. All fusions were investigated in wild-type and PHB-negative backgrounds. Colocalization with PHB granules was found for all PhaC variants and for PhaP1 to PhaP5. Additionally, eYFP fusions with H16_B1988 and H16_B2326 colocalized with PHB. Fusions of H16_B2296 with eYFP, however, did not colocalize with PHB granules but did colocalize with the nucleoid region. Notably, all fusions (except H16_B2296) were soluble in a ΔphaC1 strain. These data confirm that H16_B1988 and H16_B2326 but not H16_B2296 encode true PGAPs, for which we propose the designation PhaP6 (H16_B1988) and PhaP7 (H16_B2326). When localization of phasins was investigated at different stages of PHB accumulation, fusions of PhaP6 and PhaP7 were soluble in the first 3 h under PHB-permissive conditions, although PHB granules appeared after 10 min. At later time points, the fusions colocalized with PHB. Remarkably, PHB granules of strains expressing eYFP fusions with PhaP5, PhaP6, or PhaP7 localized predominantly near the cell poles or in the area of future septum formation. This phenomenon was not observed for the other PGAPs (PhaP1 to PhaP4, PhaC1, PhaC1-C319A, and PhaC2) and indicated that some phasins

Processing and characterization of solid and microcellular PHBV/PBAT blend and its RWF/nanoclay composites

Science.gov (United States)

Alireza Javadi; Yottha Srithep; Jungjoo Lee; Srikanth Pilla; Craig Clemons; Shaoqin Gong; Lih-Sheng Turng

2010-01-01

Solid and microcellular components made of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)/ poly (butylenes adipate-co-terephthalate) (PBAT) blend (weight ration of PHBV:PBAT = 30:70), recycled wood fiber (RWF), and nanoclay (NC) were prepared via a conventional and microcellular-injection molding process, respectively. Morphology, thermal properties, and...

Combined Determination of Poly-β-Hydroxyalkanoic and Cellular Fatty Acids in Starved Marine Bacteria and Sewage Sludge by Gas Chromatography with Flame Ionization or Mass Spectrometry Detection

Science.gov (United States)

Odham, Göran; Tunlid, Anders; Westerdahl, Gunilla; Mårdén, Per

1986-01-01

Extraction of lipids from bacterial cells or sewage sludge samples followed by simple and rapid extraction procedures and room temperature esterification with pentafluorobenzylbromide allowed combined determinations of poly-β-hydroxyalkanoate constituents and fatty acids. Capillary gas chromatography and flame ionization or mass spectrometric detection was used. Flame ionization permitted determination with a coefficient of variation ranging from 10 to 27% at the picomolar level, whereas quantitative chemical ionization mass spectrometry afforded sensitivities for poly-β-hydroxyalkanoate constituuents in the attomolar range. The latter technique suggests the possibility of measuring such components in bacterial assemblies with as few as 102 cells. With the described technique using flame ionization detection, it was possible to study the rapid formation of poly-β-hydroxyalkanoate during feeding of a starved marine bacterium isolate with a complex medium or glucose and correlate the findings to changes in cell volumes. Mass spectrometric detection of short β-hydroxy acids in activated sewage sludge revealed the presence of 3-hydroxybutyric, 3-hydroxyhexanoic, and 3-hydroxyoctanoic acids in the relative proportions of 56, 5 and 39%, respectively. No odd-chain β-hydroxy acids were found. PMID:16347181

Exercise promotes the expression of brain derived neurotrophic factor (BDNF) through the action of the ketone body β-hydroxybutyrate.

Science.gov (United States)

Sleiman, Sama F; Henry, Jeffrey; Al-Haddad, Rami; El Hayek, Lauretta; Abou Haidar, Edwina; Stringer, Thomas; Ulja, Devyani; Karuppagounder, Saravanan S; Holson, Edward B; Ratan, Rajiv R; Ninan, Ipe; Chao, Moses V

2016-06-02

Exercise induces beneficial responses in the brain, which is accompanied by an increase in BDNF, a trophic factor associated with cognitive improvement and the alleviation of depression and anxiety. However, the exact mechanisms whereby physical exercise produces an induction in brain Bdnf gene expression are not well understood. While pharmacological doses of HDAC inhibitors exert positive effects on Bdnf gene transcription, the inhibitors represent small molecules that do not occur in vivo. Here, we report that an endogenous molecule released after exercise is capable of inducing key promoters of the Mus musculus Bdnf gene. The metabolite β-hydroxybutyrate, which increases after prolonged exercise, induces the activities of Bdnf promoters, particularly promoter I, which is activity-dependent. We have discovered that the action of β-hydroxybutyrate is specifically upon HDAC2 and HDAC3, which act upon selective Bdnf promoters. Moreover, the effects upon hippocampal Bdnf expression were observed after direct ventricular application of β-hydroxybutyrate. Electrophysiological measurements indicate that β-hydroxybutyrate causes an increase in neurotransmitter release, which is dependent upon the TrkB receptor. These results reveal an endogenous mechanism to explain how physical exercise leads to the induction of BDNF.

Availability of neurotransmitter glutamate is diminished when beta-hydroxybutyrate replaces glucose in cultured neurons.

Science.gov (United States)

Lund, Trine M; Risa, Oystein; Sonnewald, Ursula; Schousboe, Arne; Waagepetersen, Helle S

2009-07-01

Ketone bodies serve as alternative energy substrates for the brain in cases of low glucose availability such as during starvation or in patients treated with a ketogenic diet. The ketone bodies are metabolized via a distinct pathway confined to the mitochondria. We have compared metabolism of [2,4-(13)C]beta-hydroxybutyrate to that of [1,6-(13)C]glucose in cultured glutamatergic neurons and investigated the effect of neuronal activity focusing on the aspartate-glutamate homeostasis, an essential component of the excitatory activity in the brain. The amount of (13)C incorporation and cellular content was lower for glutamate and higher for aspartate in the presence of [2,4-(13)C]beta-hydroxybutyrate as opposed to [1,6-(13)C]glucose. Our results suggest that the change in aspartate-glutamate homeostasis is due to a decreased availability of NADH for cytosolic malate dehydrogenase and thus reduced malate-aspartate shuttle activity in neurons using beta-hydroxybutyrate. In the presence of glucose, the glutamate content decreased significantly upon activation of neurotransmitter release, whereas in the presence of only beta-hydroxybutyrate, no decrease in the glutamate content was observed. Thus, the fraction of the glutamate pool available for transmitter release was diminished when metabolizing beta-hydroxybutyrate, which is in line with the hypothesis of formation of transmitter glutamate via an obligatory involvement of the malate-aspartate shuttle.

Influence of biodegradable polymer coatings on corrosion, cytocompatibility and cell functionality of Mg-2.0Zn-0.98Mn magnesium alloy.

Science.gov (United States)

Witecka, Agnieszka; Yamamoto, Akiko; Idaszek, Joanna; Chlanda, Adrian; Święszkowski, Wojciech

2016-08-01

Four kinds of biodegradable polymers were employed to prepare bioresorbable coatings on Mg-2.0Zn-0.98Mn (ZM21) alloy to understand the relationship between polymer characteristics, protective effects on substrate corrosion, cytocompatibility and cell functionality. Poly-l-lactide (PLLA), poly(3-hydroxybutyrate) (PHB), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) or poly(lactic-co-glycolic) acid (PLGA) was spin-coated on ZM21, obtaining a smooth, non-porous coating less than 0.5μm in thickness. Polymer coating characterization, a degradation study, and biocompatibility evaluations were performed. After 4 w of immersion into cell culture medium, degradation of PLGA and PLLA coatings were confirmed by ATR-FTIR observation. The coatings of PLLA, PHB and PHBV, which have lower water permeability and slower degradation than PLGA, provide better suppression of initial ZM21 degradation and faster promotion of human osteosarcoma cell growth and differentiation. Copyright © 2016 Elsevier B.V. All rights reserved.

Optimization PHAs production from dairy industry wastewater (cheese whey) by Azohydromonas lata DSMZ 1123

OpenAIRE

M. Sharifzadeh Baei; G.D. Najafpour; Z. Lasemi; F. Tabandeh; H. Younesi; H. Issazadeh; M. Khodabandeh

2010-01-01

In the present research, whey was used as useful substrate which retained from permeates of dairy industry. The obtained whey was hydrolyzed to cleave its main carbon source, lactose to glucose and galactose.The hydrolyzed products were chosen as carbon sources for the production of poly-3-hydroxybutyric acid (PHB) by Azohydromonas lata DSMZ 1123. The biosynthesis of PHA copolyesters containing 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) units from hydrolyzed whey permeate and valerat...

Improvement of Accuracy in Flow Immunosensor System by Introduction of Poly-2-[3-(methacryloylaminopropylammonio]ethyl 3-aminopropyl Phosphate

Directory of Open Access Journals (Sweden)

Yusuke Fuchiwaki

2011-01-01

Full Text Available In order to improve the accuracy of immunosensor systems, poly-2-[3-(methacryloylaminopropylammonio]ethyl 3-aminopropyl phosphate (poly-3MAm3AP, which includes both phosphorylcholine and amino groups, was synthesized and applied to the preparation of antibody-immobilized beads. Acting as an antibody-immobilizing material, poly-3MAm3AP is expected to significantly lower nonspecific adsorption due to the presence of the phosphorylcholine group and recognize large numbers of analytes due to the increase in antibody-immobilizing sites. The elimination of nonspecific adsorption was compared between the formation of a blocking layer on antibody-immobilized beads and the introduction of a material to combine antibody with beads. Determination with specific and nonspecific antibodies was then investigated for the estimation of signal-to-noise ratio. Signal intensities with superior signal-to-noise ratios were obtained when poly-3MAm3AP was introduced. This may be due to the increase in antibody-immobilizing sites and the extended space for antigen-antibody interaction resulting from the electrostatic repulsion of poly-3MAm3AP. Thus, the application of poly-3MAm3AP coatings to immunoassay beads was able to improve the accuracy of flow immunosensor systems.

Biodegradation of films of low density polyethylene (LDPE, poly(hydroxibutyrate-co-valerate (PHBV, and LDPE/PHBV (70/30 blend with Paecilomyces variotii

Directory of Open Access Journals (Sweden)

Thayse Marques Passos

2015-02-01

Full Text Available The increased consumption of plastics in the world has been a subject of great concern and special attention by the scientific community. The aim is to promote development of materials that are biodegradable in a shorter time upon disposal in the environment. The most used synthetic plastics are difficult to biodegrade because they are made of long hydrocarbon chains, such as polyethylene (PE, polypropylene (PP, poly(vinyl chloride (PVC, which are hydrophobic and resistant to the action of microbial enzymes. The use of alternative materials (natural polyesters can minimize the harm to dumps and landfills upon their disposal, because they are susceptible to the action of microorganisms. In this study we evaluated the biodegradation/biodeterioration of PHBV (poly(3-hydroxybutyrate-co-hydroxyvalerate films, LDPE (low density polyethylene and the blend of LDPE/PHBV (70/30 by the fungus Paecilomyces variotii, using different methods: optical microscopy (OM, scanning electronic microscopy (SEM and Fourier Transform Infrared spectroscopy (FTIR.

Poly(3,4-ethylenedioxythiophene)-based glucose biosensors

NARCIS (Netherlands)

Kros, A.; Hövell, W.F.M. van; Sommerdijk, N.A.J.M.; Nolte, R.J.M.

2001-01-01

Amperometric biosensors for the recognition of glucose oxidase (GOx) based on poly(3,4-ethylenedioxythiophene) (PEDOT) were fabricated for the first time. The resulting biosensor has potential applications for long-term glucose measurements.

Dip-coating of poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) anodes for efficient polymer solar cells

International Nuclear Information System (INIS)

Huang, Like; Hu, Ziyang; Zhang, Ke; Chen, Peipei; Zhu, Yuejin

2015-01-01

The fabrication of anodes and active layers by dip-coating in indium tin oxide (ITO)-free polymer solar cells (PSCs) is investigated. A highly conductive poly(3, 4-ethylenedioxythiophene):poly(styrenesulfonate)(PEDOT:PSS) layer was used as an anode while a blend film of poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C61 butyric acid methyl ester (PCBM) was employed as an active layer. The transmittance and sheet resistance of dip-coated PEDOT:PSS layers prepared with different thickness were studied. These layers were integrated into PSCs. The PSCs with the dip-coated PEDOT:PSS and P3HT:PCBM films exhibited power conversion efficiencies of 3.21% and 3.03% on glass and polyethylene terephthalate substrates, respectively, comparable to those of conventional ITO-based cells. Our research results suggest the feasibility of fabricating PSCs without a traditional spin-coating process and the possibility to substitute the ITO electrodes for conducting polymer films using the facile dip-coating method. - Highlights: • ITO-free polymer solar cells (PSCs) were fabricated by dip coating method. • Highly conductive PEDOT:PSS films used as anode were prepared. • The ITO-free PSCs performance was comparable with that of the spin coated devices. • Our results suggest the possibility of replacing ITO with dip coated PEDOT:PSS

Dip-coating of poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) anodes for efficient polymer solar cells

Energy Technology Data Exchange (ETDEWEB)

Huang, Like; Hu, Ziyang, E-mail: huziyang@nbu.edu.cn; Zhang, Ke; Chen, Peipei; Zhu, Yuejin, E-mail: zhuyuejin@nbu.edu.cn

2015-03-02

The fabrication of anodes and active layers by dip-coating in indium tin oxide (ITO)-free polymer solar cells (PSCs) is investigated. A highly conductive poly(3, 4-ethylenedioxythiophene):poly(styrenesulfonate)(PEDOT:PSS) layer was used as an anode while a blend film of poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C61 butyric acid methyl ester (PCBM) was employed as an active layer. The transmittance and sheet resistance of dip-coated PEDOT:PSS layers prepared with different thickness were studied. These layers were integrated into PSCs. The PSCs with the dip-coated PEDOT:PSS and P3HT:PCBM films exhibited power conversion efficiencies of 3.21% and 3.03% on glass and polyethylene terephthalate substrates, respectively, comparable to those of conventional ITO-based cells. Our research results suggest the feasibility of fabricating PSCs without a traditional spin-coating process and the possibility to substitute the ITO electrodes for conducting polymer films using the facile dip-coating method. - Highlights: • ITO-free polymer solar cells (PSCs) were fabricated by dip coating method. • Highly conductive PEDOT:PSS films used as anode were prepared. • The ITO-free PSCs performance was comparable with that of the spin coated devices. • Our results suggest the possibility of replacing ITO with dip coated PEDOT:PSS.

Poly(3-hexylthiophene)/ZnO hybrid pn junctions for microelectronics applications

DEFF Research Database (Denmark)

Katsia, E.; Huby, N.; Tallarida, G.

2009-01-01

Hybrid poly(3-hexylthiophene)/ZnO devices are investigated as rectifying heterojunctions for microelectronics applications. A low-temperature atomic layer deposition of ZnO on top of poly(3-hexylthiophene) allows the fabrication of diodes featuring a rectification ratio of nearly 105 at ±4 V...

Dual-layered nanogel-coated hollow lipid/polypeptide conjugate assemblies for potential pH-triggered intracellular drug release.

Directory of Open Access Journals (Sweden)

Wen-Hsuan Chiang

Full Text Available To achieve effective intracellular anticancer drug delivery, the polymeric vesicles supplemented with the pH-responsive outlayered gels as a delivery system of doxorubicin (DOX were developed from self-assembly of the lipid/polypeptide adduct, distearin grafted poly(γ-glutamic acid (poly(γ-GA, followed by sequential deposition of chitosan and poly(γ-GA-co-γ-glutamyl oxysuccinimide-g-monomethoxy poly(ethylene glycol in combination with in situ covalent cross-linking on assembly surfaces. The resultant gel-caged polymeric vesicles (GCPVs showed superior performance in regulating drug release in response to the external pH change. Under typical physiological conditions (pH 7.4 and 37 °C at which the γ-GA/DOX ionic pairings remained mostly undisturbed, the dense outlayered gels of GCPVs significantly reduced the premature leakage of the uncomplexed payload. With the environmental pH being reduced from pH 7.4 to 4.7, the drug liberation was appreciably promoted by the massive disruption of the ionic γ-GA/DOX complexes along with the significant swelling of nanogel layers upon the increased protonation of chitosan chain segments. After being internalized by HeLa cells via endocytosis, GCPVs exhibited cytotoxic effect comparable to free DOX achieved by rapidly releasing the payload in intracellular acidic endosomes and lysosomes. This strongly implies the great promise of such unique GCPVs as an intracellular drug delivery carrier for potential anticancer treatment.

Galectin-3 guides intracellular trafficking of some human serotransferrin glycoforms

DEFF Research Database (Denmark)

Carlsson, Carl Michael; Bengtson, Per; Cucak, Helena

2013-01-01

these transferrin glycoforms differently after preloading with exogenously added galectin-3. In all, this study provides the first evidence of a functional role for transferrin glycans, in intracellular trafficking after uptake. Moreover, the galectin-3 bound glycoform increased in cancer, suggesting...

[Treatment of gamma-hydroxybutyrate withdrawal].

Science.gov (United States)

Strand, Niels August Willer; Petersen, Tonny Studsgaard; Nielsen, Lars Martin; Boegevig, Soren

2017-12-11

Gamma-hydroxybutyrate (GHB) is a drug of abuse, for which physical addiction develops quickly. GHB withdrawal can develop into a life-threatening condition and has previously been treated mainly with benzodiazepines. These have not always proven effective, leading to long hospitalizations in intensive care units. Based on successful Dutch treatment results for using GHB to treat GHB withdrawal symptoms, we propose to implement a similar method in Denmark. The method requires an interdisciplinary effort for which The Danish Poison Information Centre should be consulted for expertise.

The metabolic impact of β-hydroxybutyrate on neurotransmission: Reduced glycolysis mediates changes in calcium responses and KATP channel receptor sensitivity

DEFF Research Database (Denmark)

Lund, Trine Meldgaard; Ploug, K.B.; Iversen, Anne

2015-01-01

-hydroxybutyrate might change neuronal function as there is a known coupling between metabolism and neurotransmission. The purpose of this study was to shed light on the effects of the ketone body β-hydroxybutyrate on glycolysis and neurotransmission in cultured murine glutamatergic neurons. Previous studies have shown...... an effect of β-hydroxybutyrate on glucose metabolism, and the present study further specified this by showing attenuation of glycolysis when β-hydroxybutyrate was present in these neurons. In addition, the NMDA receptor-induced calcium responses in the neurons were diminished in the presence of β...... to a combination of glucose and R-β-hydroxybutyrate in cultured neurons. Using the latter combination, glycolysis was diminished, NMDA receptor-induced calcium responses were lower, and the KATP channel blocker glibenclamide caused a higher transmitter release....

Expression of human poly (ADP-ribose) polymerase 1 in Saccharomyces cerevisiae: Effect on survival, homologous recombination and identification of genes involved in intracellular localization

Energy Technology Data Exchange (ETDEWEB)

La Ferla, Marco; Mercatanti, Alberto; Rocchi, Giulia; Lodovichi, Samuele; Cervelli, Tiziana; Pignata, Luca [Yeast Genetics and Genomics, Institute of Clinical Physiology, National Council of Research (CNR), via Moruzzi 1, 56122 Pisa (Italy); Caligo, Maria Adelaide [Section of Genetic Oncology, University Hospital and University of Pisa, via Roma 57, 56125 Pisa (Italy); Galli, Alvaro, E-mail: alvaro.galli@ifc.cnr.it [Yeast Genetics and Genomics, Institute of Clinical Physiology, National Council of Research (CNR), via Moruzzi 1, 56122 Pisa (Italy)

2015-04-15

Highlights: • The human poly (ADP-ribose) polymerase 1 (PARP-1) gene affects growth and UV-induced homologous recombination in yeast. • PARP-1 chemical inhibition impacts yeast growth and UV-induced recombination. • A genome-wide screen identifies 99 yeast genes that suppress the growth defect inferred by PARP-1. • Bioinformatics analysis identifies 41 human orthologues that may have a role in PARP-1 intracellular localization. • The findings suggest that PARP-1 nuclear localization may affect the response to PARP inhibitors in cancer therapy. - Abstract: The poly (ADP-ribose) polymerase 1 (PARP-1) actively participates in a series of functions within the cell that include: mitosis, intracellular signaling, cell cycle regulation, transcription and DNA damage repair. Therefore, inhibition of PARP1 has a great potential for use in cancer therapy. As resistance to PARP inhibitors is starting to be observed in patients, thus the function of PARP-1 needs to be studied in depth in order to find new therapeutic targets. To gain more information on the PARP-1 activity, we expressed PARP-1 in yeast and investigated its effect on cell growth and UV induced homologous recombination. To identify candidate genes affecting PARP-1 activity and cellular localization, we also developed a yeast genome wide genetic screen. We found that PARP-1 strongly inhibited yeast growth, but when yeast was exposed to the PARP-1 inhibitor 6(5-H) phenantridinone (PHE), it recovered from the growth suppression. Moreover, we showed that PARP-1 produced PAR products in yeast and we demonstrated that PARP-1 reduced UV-induced homologous recombination. By genome wide screening, we identified 99 mutants that suppressed PARP-1 growth inhibition. Orthologues of human genes were found for 41 of these yeast genes. We determined whether the PARP-1 protein level was altered in strains which are deleted for the transcription regulator GAL3, the histone H1 gene HHO1, the HUL4 gene, the

Investigation of Poly(3,4-ethylenedioxythiophene:Poly(styrenesulfonate Hole Transport Layer for Solution-Processed Polymer Solar Cells

Directory of Open Access Journals (Sweden)

Chengxi Zhang

2015-01-01

Full Text Available The inverted polymer solar cell was prepared by self-made spray-coating system, and the poly(3,4-ethylenedioxythiophene:poly(styrenesulfonate (PEDOT:PSS hole transport layer was studied. 220 nm poly-(3-hexylthiophene:[6,6]-phenylC61butyric-acid methyl-ester (P3HT:PCBM and 40 nm PEDOT:PSS were deposited on ZnO thin film subsequently by solution spray coating. Different volume of isopropyl alcohol was introduced into PEDOT:PSS to decrease the contact angle and obtain the optimum Marangoni flow. The surface morphology and roughness of PEDOT:PSS films were characterized by atomic force microscopy with varied deposition temperature from 70°C to 160°C. The improvement of power conversion efficiency (PCE was attributed to the enhancement of vertical phase separation in PEDOT:PSS film, which improved the charge transfer in the bulk cell. The highest PCE of spray-coated PSCs reached 2.80% after postannealing for 10 min.

Optimized methods to measure acetoacetate, 3-hydroxybutyrate, glycerol, alanine, pyruvate, lactate and glucose in human blood using a centrifugal analyser with a fluorimetric attachment

OpenAIRE

Stappenbeck, R.; Hodson, A. W.; Skillen, A. W.; Agius, L.; Alberti, K. G. M. M.

1990-01-01

Optimized methods are described for the analysis of glucose, lactate, pyruvate, alanine, glycerol, D-3-hydroxybutyrate and acetoacetate in perchloric acid extracts of human blood using the Cobas Bio centrifugal analyser. Glucose and lactate are measured using the photometric mode and other metabolites using the fluorimetric mode. The intra-assay coefficients of variation ranged from 0.7 to 4.1%, except with very low levels of pyruvate and acetoacetate where the coefficients of variation were ...

Effect of ageing time on mechanical properties of plasticized poly(hydroxybutyrate) (PHB)

Science.gov (United States)

Farris, Giuseppe; Cinelli, Patrizia; Anguillesi, Irene; Salvadori, Sara; Coltelli, Maria-Beatrice; Lazzeri, Andrea

2014-05-01

Polyhydroxybutyrate (PHB) based materials were prepared by melt extrusion by using different plasticizers, such as poly(ethylene glycol)s (PEG)s having different molecular weight (400, 1500 and 4000). The plasticizers content was varied in the range 10-20% by weight versus the PHB polymeric matrix. The variation of tensile properties of the different samples was monitored as a function of time of ageing to study the stability of the material. The elastic modulus and tensile strength increased as a function of time, whereas the strain at break decreased. The experimental results were explained by considering both the demixing of the plasticizers and the occurring of secondary crystallization. Moreover the variation in mechanical properties was correlated to the structure and concentration of the different plasticizers employed.

PLGA and PHBV Microsphere Formulations and Solid-State Characterization

DEFF Research Database (Denmark)

Yang, Chiming; Plackett, David; Needham, David

2009-01-01

To develop and characterize the solid-state properties of poly(DL-lactic-co-glycolic acid) (PLGA) and poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) microspheres for the localized and controlled release of fusidic acid (FA). The effects of FA loading and polymer composition on the me...... of a DCM-FA-rich phase in the forming microsphere....

Unusual poly(3-hydroxyalkanoate) (PHA) biosynthesis behavior of Pseudomonas putida Bet001 and Delftia tsuruhatensis Bet002 isolated from palm oil mill effluent.

Science.gov (United States)

Razaif-Mazinah, Mohd Rafais Mohd; Anis, Siti Nor Syairah; Harun, Hazwani Izzati; Rashid, Khairunnisa Abdul; Annuar, Mohamad Suffian Mohamad

2017-03-01

Pseudomonas putida Bet001 and Delftia tsuruhatensis Bet002, isolated from palm oil mill effluent, accumulated poly(3-hydroxyalkanoates) (PHAs) when grown on aliphatic fatty acids, sugars, and glycerol. The substrates were supplied at 20:1 C/N mole ratio. Among C-even n-alkanoic acids, myristic acid gave the highest PHA content 26 and 28 wt% in P. putida and D. tsuruhatensis, respectively. Among C-odd n-alkanoic acids, undecanoic gave the highest PHA content at 40 wt% in P. putida and 46 wt% in D. tsuruhatensis on pentadecanoic acid. Sugar and glycerol gave <10 wt% of PHA content for both bacteria. Interestingly, D. tsuruhatensis accumulated both short- and medium-chain length PHA when supplied with n-alkanoic acids ranging from octanoic to lauric, sucrose, and glycerol with 3-hydroxybutyrate as the major monomer unit. In P. putida, the major hydroxyalkanoates unit was 3-hydroxyoctanoate and 3-hydroxydecanoate when grown on C-even acids. Conversely, 3-hydroxyheptanoate, 3-hydrxoynonanoate, and 3-hydroxyundecanoate were accumulated with C-odd acids. Weight-averaged molecular weight (M w ) was in the range of 53-81 kDa and 107-415 kDa for P. putida and D. tsuruhatensis, respectively. Calorimetric analyses indicated that both bacteria synthesized semicrystalline polymer with good thermal stability with degradation temperature (T d ) ranging from 178 to 282 °C. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Glyphosate and AMPA inhibit cancer cell growth through inhibiting intracellular glycine synthesis.

Science.gov (United States)

Li, Qingli; Lambrechts, Mark J; Zhang, Qiuyang; Liu, Sen; Ge, Dongxia; Yin, Rutie; Xi, Mingrong; You, Zongbing

2013-01-01

Glycine is a nonessential amino acid that is reversibly converted from serine intracellularly by serine hydroxymethyltransferase. Glyphosate and its degradation product, aminomethylphosphonic acid (AMPA), are analogs to glycine, thus they may inhibit serine hydroxymethyltransferase to decrease intracellular glycine synthesis. In this study, we found that glyphosate and AMPA inhibited cell growth in eight human cancer cell lines but not in two immortalized human normal prostatic epithelial cell lines. AMPA arrested C4-2B and PC-3 cancer cells in the G1/G0 phase and inhibited entry into the S phase of the cell cycle. AMPA also promoted apoptosis in C4-2B and PC-3 cancer cell lines. AMPA upregulated p53 and p21 protein levels as well as procaspase 9 protein levels in C4-2B cells, whereas it downregulated cyclin D3 protein levels. AMPA also activated caspase 3 and induced cleavage of poly (adenosine diphosphate [ADP]-ribose) polymerase. This study provides the first evidence that glyphosate and AMPA can inhibit proliferation and promote apoptosis of cancer cells but not normal cells, suggesting that they have potentials to be developed into a new anticancer therapy.

Thermal, mechanical and morphological characterization of plasticized PLA-PHB blends

Science.gov (United States)

A blend of poly(lactic acid) (PLA) (75% by weight) and poly(3-hydroxybutyrate) (PHB) (25% by weight) with a polyester plasticizer (Lapol 108) at two different concentrations (5 and 7% by weight per 100 parts of the blends) were investigated by TGA, DSC, XRD, SEM, mechanical testing and biodegradatio...

Utilization of D-beta-hydroxybutyrate and oleate as alternate energy fuels in brain cell cultures of newborn mice after hypoxia at different glucose concentrations.

Science.gov (United States)

Bossi, E; Kohler, E; Herschkowitz, N

1989-11-01

In dissociated whole brain cell cultures from newborn mice, we have previously shown that during glucose deprivation under normoxia, D-beta-hydroxybutyrate and oleic acid are increasingly used for energy production. We now asked whether this glucose dependency of the utilization of D-beta-hydroxybutyrate and oleic acid as alternate energy fuels is also present after a hypoxic phase. 3-Hydroxy[3-14C]butyrate or [U-14C]oleic acid were added to 7- and 14-d-old cultures and 14CO2-production compared after hypoxia in normal and glucose-deprived conditions. After hypoxia, the ability of the cells 7 d in culture to increase D-beta-hydroxybutyrate consumption in response to glucose deprivation is diminished, 14-d-old cells lose this ability. In contrast, after hypoxia, both 7- and 14-d-old cultures maintain or even improve the ability to increase oleate consumption, when glucose is lacking.

Synthesis of Poly(3,4-Ethylenedioxy thiophene)-Poly(Styrene-4-Sulfonate) Composites for Support Fuel Cell Catalyst Layer

International Nuclear Information System (INIS)

Eko Sulistiyono; Murni Handayani

2009-01-01

Synthesis of poly(3,4-ethylenedioxy thiophene)-poly(styrene-4-sulfonate) composites for support fuel cell catalyst layer are synthesis composites which become fuel cell catalyst support so that catalyst has optimal performance. Main function of composites is support platinum particle for application in fuel cell. This article explains the result of composites production process from ( 3,4 Ethylenedioxy thiophene) and Sodium poly( styrene - 4-sulfonate) using two methods Jingning Shan method (method 1) and Zhigang Qi and Peter G.Pickup method (method 2). Analysis of the synthesis results used Scanning Electron Microscopic –Electron Dispersive X – Ray Spectrophotometer (SEM-EDS ). The analysis result show that both methods produce polymer agglomerate into a sponge-like morphology. Composite from method 1 has morphology, pores and proton transport better than composite produced by method 2. (author)

Reliability of poly 3,4-ethylenedioxythiophene strain gauge

DEFF Research Database (Denmark)

Mateiu, Ramona Valentina; Lillemose, Michael; Hansen, Thomas Steen

2007-01-01

We report on the experimentally observed reliability of the piezoresistive effect in strained poly 3,4-ethylenedioxythiophene (PEDT). PEDT is an intrinsic conductive polymer which can be patterned by conventional Cleanroom processing, and thus presents a promising material for all-polymer Microsy......We report on the experimentally observed reliability of the piezoresistive effect in strained poly 3,4-ethylenedioxythiophene (PEDT). PEDT is an intrinsic conductive polymer which can be patterned by conventional Cleanroom processing, and thus presents a promising material for all......-polymer Microsystems. The measurements are made on microfabricated test chips with PEDT resistors patterned by conventional UV-lithography and reactive ion etching (RIE). We determine a gauge factor of 3.41 ± 0.42 for the strained PEDT and we see an increase in resistivity from 1.98 · 104 X m to 2.22 · 104 X m when...

Tough, semiconducting polyethylene-poly(3-hexylthiophene) diblock copolymers

DEFF Research Database (Denmark)

Müller, C.; Goffri, S.; Breiby, Dag Werner

2007-01-01

Semiconducting diblock copolymers of polyethylene (PE) and regioregular poly(3-hexylthiophene) (P3HT) are demonstrated to exhibit a rich phase behaviour, judicious use of which permitted us to fabricate field-effect transistors that show saturated charge carrier mobilities, mu(FET), as high as 2 x...

Quantification of the Force of Nanoparticle-Cell Membrane Interactions and Its Influence on Intracellular Trafficking of Nanoparticles

Science.gov (United States)

Vasir, Jaspreet K.; Labhasetwar, Vinod

2008-01-01

Understanding the interaction of nanoparticles (NPs) with the cell membrane and their trafficking through cells is imperative to fully explore the use of NPs for efficient intracellular delivery of therapeutics. Here, we report a novel method of measuring the force of NP-cell membrane interactions using atomic force microscopy (AFM). Poly(dl-lactide co-glycolide, PLGA) NPs functionalized with poly-l-lysine were used as a model system, to demonstrate that this force determines the adhesive interaction of NPs with the cell membrane and in turn the extent of cellular uptake of NPs, and hence that of the encapsulated therapeutic. Cellular uptake of NPs was monitored using AFM imaging, and the dynamics of their intracellular distribution was quantified using confocal microscopy. Results demonstrated that the functionalized NPs have a five-fold greater force of adhesion with the cell membrane and the time-lapse AFM images show their rapid internalization than unmodified NPs. The intracellular trafficking study showed that the functionalized NPs escape more rapidly and efficiently from late endosomes than unmodified NPs and result in 10-fold higher intracellular delivery of the encapsulated model protein. The findings described herein enhance our basic understanding of the NP-cell membrane interaction on the basis of physical phenomena that could have wider applications in developing efficient nanocarrier systems for intracellular delivery of therapeutics. PMID:18692238

Conditions With High Intracellular Glucose Inhibit Sensing Through Glucose Sensor Snf3 in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Karhumaa, Kaisa; Wu, B.Q.; Kielland-Brandt, Morten

2010-01-01

as for amino acids. An alternating-access model of the function of transporter-like sensors has been previously suggested based on amino acid sensing, where intracellular ligand inhibits binding of extracellular ligand. Here we studied the effect of intracellular glucose on sensing of extracellular glucose...... through the transporter-like sensor Snf3 in yeast. Sensing through Snf3 was determined by measuring degradation of Mth1 protein. High intracellular glucose concentrations were achieved by using yeast strains lacking monohexose transporters which were grown on maltose. The apparent affinity...... of extracellular glucose to Snf3 was measured for cells grown in non-fermentative medium or on maltose. The apparent affinity for glucose was lowest when the intracellular glucose concentration was high. The results conform to an alternating-access model for transporter-like sensors. J. Cell. Biochem. 110: 920...

Intracellular Trafficking Modulation by Ginsenoside Rg3 Inhibits Brucella abortus Uptake and Intracellular Survival within RAW 264.7 Cells.

Science.gov (United States)

Huy, Tran Xuan Ngoc; Reyes, Alisha Wehdnesday Bernardo; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, WonGi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee; Kim, Suk

2017-03-28

Ginsenoside Rg3, a saponin extracted from ginseng, has various pharmacological and biological activities; however, its effects against Brucella infection are still unclear. Herein, the inhibitory effects of ginsenoside Rg3 against intracellular parasitic Brucella infection were evaluated through bacterial infection, adherence assays, and LAMP-1 colocalization, as well as immunoblotting and FACS for detecting MAPK signaling proteins and F-actin polymerization, respectively. The internalization, intracellular growth, and adherence of Brucella abortus in Rg3-treated RAW 264.7 cells were significantly decreased compared with the Rg3-untreated control. Furthermore, an apparent reduction of F-actin content and intensity of F-actin fluorescence in Rg3-treated cells was observed compared with B. abortus -infected cells without treatment by flow cytometry analysis and confocal microscopy, respectively. In addition, treating cells with Rg3 decreased the phosphorylation of MAPK signaling proteins such as ERK 1/2 and p38 compared with untreated cells. Moreover, the colocalization of B. abortus -containing phagosomes with LAMP-1 was markedly increased in Rg3-treated cells. These findings suggest that ginsenoside Rg3 inhibits B. abortus infection in mammalian cells and can be used as an alternative approach in the treatment of brucellosis.

Exploring proton doping in poly-3-methylpyrrole by infrared spectroscopy

International Nuclear Information System (INIS)

Sanchez de la Blanca, E.; Carrillo, I.; Redondo, M.I.; Gonzalez-Tejera, M.J.; Garcia, M.V.

2007-01-01

Structural changes induced by electrochemical redox processes or by pH variations on conducting poly-3-methylpyrrole, electrochemically synthesized in NaClO 4 acetonitrile solution, have been studied by infrared spectroscopy. With this aim infrared spectra of perchlorate doped poly-3-methylpyrrole films at different oxidation states as well as after immersion in acid (pH = 1) and basic (pH = 12.6) aqueous solutions have been analysed. The existence of proton-doping mechanism in this polymer has been confirmed from the comparative study of spectra of oxidized/reduced and acid/basic treated polymer

Unraveling the Order and Disorder in Poly(3,4-ethylenedioxythiophene)/Poly(styrenesulfonate) Nanofilms

KAUST Repository

Zhou, Jian; Anjum, Dalaver H.; Lubineau, Gilles; Li, Erqiang; Thoroddsen, Sigurdur T

2015-01-01

Conductive polymer poly(3,4-ethylenedioxythiophene)/poly(styrenesulfonate) (PEDOT/PSS) exhibits a tunable conductivity ranging from 0.1 to 4380 S·cm–1 under different doping and/or dedoping strategies. However, the dependence of macroscopic electrical properties on the evolution of the microstructure is not clearly understood. This is the first study that systematically investigated the spatial arrangement of the ordered and disordered phases in PEDOT/PSS nanofilms by bright-field (BF), high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) combined with electron energy loss spectroscopy (EELS) and element-thickness mapping. Our observations clarify how amorphous PSS hinders electrical transport at various length scales in the PEDOT/PSS films. Moreover, the mechanism for an enhancement in 3 orders of magnitude in electrical conductivity was proved by TEM investigation, which is mainly due to a more uniform dispersion by dedoping that opens PEDOT nanoparticle clusters in PEDOT/PSS films. Our microstructural and electrical studies show that the change in spatial arrangement and interaction of small PEDOT domains plays a considerable role in the final electron transport.

Unraveling the Order and Disorder in Poly(3,4-ethylenedioxythiophene)/Poly(styrenesulfonate) Nanofilms

KAUST Repository

Zhou, Jian

2015-08-03

Conductive polymer poly(3,4-ethylenedioxythiophene)/poly(styrenesulfonate) (PEDOT/PSS) exhibits a tunable conductivity ranging from 0.1 to 4380 S·cm–1 under different doping and/or dedoping strategies. However, the dependence of macroscopic electrical properties on the evolution of the microstructure is not clearly understood. This is the first study that systematically investigated the spatial arrangement of the ordered and disordered phases in PEDOT/PSS nanofilms by bright-field (BF), high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) combined with electron energy loss spectroscopy (EELS) and element-thickness mapping. Our observations clarify how amorphous PSS hinders electrical transport at various length scales in the PEDOT/PSS films. Moreover, the mechanism for an enhancement in 3 orders of magnitude in electrical conductivity was proved by TEM investigation, which is mainly due to a more uniform dispersion by dedoping that opens PEDOT nanoparticle clusters in PEDOT/PSS films. Our microstructural and electrical studies show that the change in spatial arrangement and interaction of small PEDOT domains plays a considerable role in the final electron transport.

The metabolic impact of β-hydroxybutyrate on neurotransmission: Reduced glycolysis mediates changes in calcium responses and KATP channel receptor sensitivity.

Science.gov (United States)

Lund, Trine M; Ploug, Kenneth B; Iversen, Anne; Jensen, Anders A; Jansen-Olesen, Inger

2015-03-01

Glucose is the main energy substrate for neurons, and ketone bodies are known to be alternative substrates. However, the capacity of ketone bodies to support different neuronal functions is still unknown. Thus, a change in energy substrate from glucose alone to a combination of glucose and β-hydroxybutyrate might change neuronal function as there is a known coupling between metabolism and neurotransmission. The purpose of this study was to shed light on the effects of the ketone body β-hydroxybutyrate on glycolysis and neurotransmission in cultured murine glutamatergic neurons. Previous studies have shown an effect of β-hydroxybutyrate on glucose metabolism, and the present study further specified this by showing attenuation of glycolysis when β-hydroxybutyrate was present in these neurons. In addition, the NMDA receptor-induced calcium responses in the neurons were diminished in the presence of β-hydroxybutyrate, whereas a direct effect of the ketone body on transmitter release was absent. However, the presence of β-hydroxybutyrate augmented transmitter release induced by the KATP channel blocker glibenclamide, thus giving an indirect indication of the involvement of KATP channels in the effects of ketone bodies on transmitter release. Energy metabolism and neurotransmission are linked and involve ATP-sensitive potassium (KATP ) channels. However, it is still unclear how and to what degree available energy substrate affects this link. We investigated the effect of changing energy substrate from only glucose to a combination of glucose and R-β-hydroxybutyrate in cultured neurons. Using the latter combination, glycolysis was diminished, NMDA receptor-induced calcium responses were lower, and the KATP channel blocker glibenclamide caused a higher transmitter release. © 2014 International Society for Neurochemistry.

Surface sulfonamide modification of poly(N-isopropylacrylamide)-based block copolymer micelles to alter pH and temperature responsive properties for controlled intracellular uptake.

Science.gov (United States)

Cyphert, Erika L; von Recum, Horst A; Yamato, Masayuki; Nakayama, Masamichi

2018-06-01

Two different surface sulfonamide-functionalized poly(N-isopropylacrylamide)-based polymeric micelles were designed as pH-/temperature-responsive vehicles. Both sulfadimethoxine- and sulfamethazine-surface functionalized micelles were characterized to determine physicochemical properties, hydrodynamic diameters, zeta potentials, temperature-dependent size changes, and lower critical solution temperatures (LCST) in both pH 7.4 and 6.8 solutions (simulating both physiological and mild low pH conditions), and tested in the incorporation of a proof-of-concept hydrophobic antiproliferative drug, paclitaxel. Cellular uptake studies were conducted using bovine carotid endothelial cells and fluorescently labeled micelles to evaluate if there was enhanced cellular uptake of the micelles in a low pH environment. Both variations of micelles showed enhanced intracellular uptake under mildly acidic (pH 6.8) conditions at temperatures slightly above their LCST and minimal uptake at physiological (pH 7.4) conditions. Due to the less negative zeta potential of the sulfamethazine-surface micelles compared to sulfadimethoxine-surface micelles, and the proximity of their LCST to physiological temperature (37°C), the sulfamethazine variation was deemed more amenable for clinically relevant temperature and pH-stimulated applications. Nevertheless, we believe both polymeric micelle variations have the capacity to be implemented as an intracellular drug or gene delivery system in response to mildly acidic conditions. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1552-1560, 2018. © 2018 Wiley Periodicals, Inc.

Intracellular trafficking of superparamagnetic iron oxide nanoparticles conjugated with TAT peptide: 3-dimensional electron tomography analysis

International Nuclear Information System (INIS)

Nair, Baiju G.; Fukuda, Takahiro; Mizuki, Toru; Hanajiri, Tatsuro; Maekawa, Toru

2012-01-01

Highlights: ► We study the intracellular localisation of TAT-SPIONs using 3-D electron tomography. ► 3-D images of TAT-SPIONs in a cell are clearly shown. ► Release of TAT-SPIONs from endocytic vesicles into the cytoplasm is clearly shown. -- Abstract: Internalisation of nanoparticles conjugated with cell penetrating peptides is a promising approach to various drug delivery applications. Cell penetrating peptides such as transactivating transcriptional activator (TAT) peptides derived from HIV-1 proteins are effective intracellular delivery vectors for a wide range of nanoparticles and pharmaceutical agents thanks to their amicable ability to enter cells and minimum cytotoxicity. Although different mechanisms of intracellular uptake and localisation have been proposed for TAT conjugated nanoparticles, it is necessary to visualise the particles on a 3-D plane in order to investigate the actual intracellular uptake and localisation. Here, we study the intracellular localisation and trafficking of TAT peptide conjugated superparamagnetic ion oxide nanoparticles (TAT-SPIONs) using 3-D electron tomography. 3-D tomograms clearly show the location of TAT-SPIONs in a cell and their slow release from the endocytic vesicles into the cytoplasm. The present methodology may well be utilised for further investigations of the behaviours of nanoparticles in cells and eventually for the development of nano drug delivery systems.

Effects of beta-hydroxybutyrate and isoproterenol on lipolysis in isolated adipocytes from periparturient dairy cows and cows with clinical ketosis

NARCIS (Netherlands)

van der Drift, S. G. A.; Everts, R. R.; Houweling, M.; van Leengoed, L. A. M. G.; Stegeman, J. A.; Tielens, A. G. M.; Jorritsma, R.

An in vitro model was used to investigate effects of beta-hydroxybutyrate and isoproterenol (beta-adrenergic receptor agonist) on lipolysis in isolated adipocytes from late pregnant and recently calved dairy cows (n = 5) and cows with clinical ketosis (n =3). Incubation with 3.0 mmol/L

Comparative analysis of poly-glycolic acid-based hybrid polymer starter matrices for in vitro tissue engineering.

Science.gov (United States)

Generali, Melanie; Kehl, Debora; Capulli, Andrew K; Parker, Kevin K; Hoerstrup, Simon P; Weber, Benedikt

2017-10-01

Biodegradable scaffold matrixes form the basis of any in vitro tissue engineering approach by acting as a temporary matrix for cell proliferation and extracellular matrix deposition until the scaffold is replaced by neo-tissue. In this context several synthetic polymers have been investigated, however a concise systematic comparative analyses is missing. Therefore, the present study systematically compares three frequently used polymers for the in vitro engineering of extracellular matrix based on poly-glycolic acid (PGA) under static as well as dynamic conditions. Ultra-structural analysis was used to examine the polymers structure. For tissue engineering (TE) three human fibroblast cell lines were seeded on either PGA-poly-4-hydroxybutyrate (P4HB), PGA-poly-lactic acid (PLA) or PGA-poly-caprolactone (PCL) patches. These patches were analyzed after 21days of culture qualitative by histology and quantitative by determining the amount of DNA, glycosaminoglycan and hydroxyproline. We found that PGA-P4HB and PGA-PLA scaffolds enhance tissue formation significantly higher than PGA-PCL scaffolds (p<0.05). Polymer remnants were visualized by polarization microscopy. In addition, biomechanical properties of the tissue engineered patches were determined in comparison to native tissue. This study may allow future studies to specifically select certain polymer starter matrices aiming at specific tissue properties of the bioengineered constructs in vitro. Copyright © 2017 Elsevier B.V. All rights reserved.

Poly(3,4-ethylenedioxythiophene)-Poly(styrenesulfonate) Interlayer Insertion Enables Organic Quaternary Memory.

Science.gov (United States)

Cheng, Xue-Feng; Hou, Xiang; Qian, Wen-Hu; He, Jing-Hui; Xu, Qing-Feng; Li, Hua; Li, Na-Jun; Chen, Dong-Yun; Lu, Jian-Mei

2017-08-23

Herein, for the first time, quaternary resistive memory based on an organic molecule is achieved via surface engineering. A layer of poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT-PSS) was inserted between the indium tin oxide (ITO) electrode and the organic layer (squaraine, SA-Bu) to form an ITO/PEDOT-PSS/SA-Bu/Al architecture. The modified resistive random-access memory (RRAM) devices achieve quaternary memory switching with the highest yield (∼41%) to date. Surface morphology, crystallinity, and mosaicity of the deposited organic grains are greatly improved after insertion of a PEDOT-PSS interlayer, which provides better contacts at the grain boundaries as well as the electrode/active layer interface. The PEDOT-PSS interlayer also reduces the hole injection barrier from the electrode to the active layer. Thus, the threshold voltage of each switching is greatly reduced, allowing for more quaternary switching in a certain voltage window. Our results provide a simple yet powerful strategy as an alternative to molecular design to achieve organic quaternary resistive memory.

Poly(I:C) induces expressions of MMP-1, -2, and -3 through various signaling pathways including IRF3 in human skin fibroblasts.

Science.gov (United States)

Yao, Cheng; Lee, Dong Hun; Oh, Jang-Hee; Kim, Min-Kyoung; Kim, Kyu Han; Park, Chi-Hyun; Chung, Jin Ho

2015-10-01

Ultraviolet (UV) irradiation can result in premature skin aging (photoaging) which is characterized by decreased expression of collagen and increased expression of matrix metalloproteinases (MMPs). Double-stranded RNAs (dsRNAs) can be generated at various conditions including virally infected cells or UV-damaged skin cells. Recent studies have shown that a synthetic dsRNA, polyinosinic-polycytidylic acid (poly(I:C)), can reduce procollagen expression in human skin fibroblasts. However, little is known about the effect of poly(I:C) on the expression of MMPs in skin fibroblasts and its underlying mechanisms. We examined the effect of poly(I:C) on MMP-1, -2, and -3 expressions in human skin fibroblasts. Then, we further explored the underlying signaling pathways involved in the processes. Human skin fibroblasts were treated with poly(I:C) for the indicated times in the presence or the absence of various chemical inhibitors or small interfering RNAs (siRNAs) at the indicated concentrations. Protein and mRNA levels of various target molecules were examined by Western blotting and quantitative real-time PCR, respectively. Poly(I:C) induced MMP-1, -2, and -3 expressions, which were dependent on TLR3. Poly(I:C) also induced activations of the mitogen-activated protein kinases (MAPKs), the nuclear factor-kappaB (NF-κB) and the interferon regulatory factor 3 (IRF3) pathways. By using specific inhibitors, we found that poly(I:C)-induced expressions of MMP-1, -2, and -3 were differentially regulated by these signaling pathways. In particular, we found that the inhibition of IRF3 signaling pathways attenuated poly(I:C)-induced expressions of all the three MMPs. Our data show that the expressions of MMP-1, -2, and -3 are induced by poly(I:C) through various signaling pathways in human skin fibroblasts and suggest that TLR3 and/or IRF3 may be good targets for regulating the expressions of MMP-1, -2, and -3 induced by dsRNAs. Copyright © 2015 Elsevier Ireland Ltd. All rights

Intracellular trafficking of superparamagnetic iron oxide nanoparticles conjugated with TAT peptide: 3-dimensional electron tomography analysis

Energy Technology Data Exchange (ETDEWEB)

Nair, Baiju G.; Fukuda, Takahiro; Mizuki, Toru; Hanajiri, Tatsuro [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan); Maekawa, Toru, E-mail: maekawa@toyo.jp [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan)

2012-05-18

Highlights: Black-Right-Pointing-Pointer We study the intracellular localisation of TAT-SPIONs using 3-D electron tomography. Black-Right-Pointing-Pointer 3-D images of TAT-SPIONs in a cell are clearly shown. Black-Right-Pointing-Pointer Release of TAT-SPIONs from endocytic vesicles into the cytoplasm is clearly shown. -- Abstract: Internalisation of nanoparticles conjugated with cell penetrating peptides is a promising approach to various drug delivery applications. Cell penetrating peptides such as transactivating transcriptional activator (TAT) peptides derived from HIV-1 proteins are effective intracellular delivery vectors for a wide range of nanoparticles and pharmaceutical agents thanks to their amicable ability to enter cells and minimum cytotoxicity. Although different mechanisms of intracellular uptake and localisation have been proposed for TAT conjugated nanoparticles, it is necessary to visualise the particles on a 3-D plane in order to investigate the actual intracellular uptake and localisation. Here, we study the intracellular localisation and trafficking of TAT peptide conjugated superparamagnetic ion oxide nanoparticles (TAT-SPIONs) using 3-D electron tomography. 3-D tomograms clearly show the location of TAT-SPIONs in a cell and their slow release from the endocytic vesicles into the cytoplasm. The present methodology may well be utilised for further investigations of the behaviours of nanoparticles in cells and eventually for the development of nano drug delivery systems.

Preparation of graphene oxide/poly (3,4-ethylenedioxytriophene): Poly (styrene sulfonate) (PEDOT:PSS) electrospun nanofibers

International Nuclear Information System (INIS)

Efelina, Vita; Widianto, Eri; Rusdiana, Dadi; Nugroho, A. A.; Kusumaatmaja, Ahmad; Triyana, Kuwat; Santoso, Iman

2016-01-01

Graphene oxide (GO)/Poly (3,4-Ethylenedioxytriophene):Poly (styrene Sulfonate) (PEDOT:PSS) nanofibers have been successfully fabricated by a simple electrospinning technique to develop conductive nanofibers with polyvinyl alcohol (PVA) act as a carrier solution. Graphene oxide has been synthesized by Hummer’s method and has been confirmed by Raman Spectroscopy, FTIR and UV-Vis Spectroscopy. GO/PEDOT:PSS composite nanofibers. The structural and morphological properties were characterized by Scanning Electron Microscopy (SEM). The result of SEM show that GO/PEDOT:PSS nanofibers has a relatively uniform morphology nanofiber with diameter between 180 nm - 340 nm with smooth nanofiber surface. The produced nanofibers from this study can be utilized for various applications such as flexible, conductive and transparent electrode.

Preparation of graphene oxide/poly (3,4-ethylenedioxytriophene): Poly (styrene sulfonate) (PEDOT:PSS) electrospun nanofibers

Energy Technology Data Exchange (ETDEWEB)

Efelina, Vita; Widianto, Eri [Department of Physics, Universitas Gadjah Mada, Sekip Utara BLS.21 Yogyakarta, 55281 Indonesia (Indonesia); Rusdiana, Dadi [Department of Physical Education, Universitas Pendidikan Indonesia, Bandung (Indonesia); Nugroho, A. A. [Department of Physics, Institut Teknologi Bandung, Jl. Ganesha 10 Bandung (Indonesia); Kusumaatmaja, Ahmad; Triyana, Kuwat; Santoso, Iman, E-mail: iman.santoso@ugm.ac.id [Department of Physics, Universitas Gadjah Mada, Sekip Utara BLS.21 Yogyakarta, 55281 Indonesia (Indonesia); Nanomaterials Research Group, Universitas Gadjah Mada,, Sekip Utara, Yogyakarta (Indonesia)

2016-04-19

Graphene oxide (GO)/Poly (3,4-Ethylenedioxytriophene):Poly (styrene Sulfonate) (PEDOT:PSS) nanofibers have been successfully fabricated by a simple electrospinning technique to develop conductive nanofibers with polyvinyl alcohol (PVA) act as a carrier solution. Graphene oxide has been synthesized by Hummer’s method and has been confirmed by Raman Spectroscopy, FTIR and UV-Vis Spectroscopy. GO/PEDOT:PSS composite nanofibers. The structural and morphological properties were characterized by Scanning Electron Microscopy (SEM). The result of SEM show that GO/PEDOT:PSS nanofibers has a relatively uniform morphology nanofiber with diameter between 180 nm - 340 nm with smooth nanofiber surface. The produced nanofibers from this study can be utilized for various applications such as flexible, conductive and transparent electrode.

A pH and Redox Dual Responsive 4-Arm Poly(ethylene glycol-block-poly(disulfide histamine Copolymer for Non-Viral Gene Transfection in Vitro and in Vivo

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Kangkang An

2014-05-01

Full Text Available A novel 4-arm poly(ethylene glycol-b-poly(disulfide histamine copolymer was synthesized by Michael addition reaction of poly(ethylene glycol (PEG vinyl sulfone and amine-capped poly(disulfide histamine oligomer, being denoted as 4-arm PEG-SSPHIS. This copolymer was able to condense DNA into nanoscale polyplexes (<200 nm in average diameter with almost neutral surface charge (+(5–10 mV. Besides, these polyplexes were colloidal stable within 4 h in HEPES buffer saline at pH 7.4 (physiological environment, but rapidly dissociated to liberate DNA in the presence of 10 mM glutathione (intracellular reducing environment. The polyplexes also revealed pH-responsive surface charges which markedly increased with reducing pH values from 7.4–6.3 (tumor microenvironment. In vitro transfection experiments showed that polyplexes of 4-arm PEG-SSPHIS were capable of exerting enhanced transfection efficacy in MCF-7 and HepG2 cancer cells under acidic conditions (pH 6.3–7.0. Moreover, intravenous administration of the polyplexes to nude mice bearing HepG2-tumor yielded high transgene expression largely in tumor rather other normal organs. Importantly, this copolymer and its polyplexes had low cytotoxicity against the cells in vitro and caused no death of the mice. The results of this study indicate that 4-arm PEG-SSPHIS has high potential as a dual responsive gene delivery vector for cancer gene therapy.

Improving the Performances of Poly(vinylphosphonic acid) by Compositing or Copolymerization with Poly(4-(α-methyl)vinyl-1H-1,2,3-triazole)

International Nuclear Information System (INIS)

Han, Shuaiyuan; Yue, Baohua; Yan, Liuming

2014-01-01

Graphical abstract: - Highlights: • Poly(4-(α-methyl)vinyl-1H-1,2,3-triazole) is synthesized • PVPA/PMVTri polymeric composite proton conducting membranes are prepared • The proton conductivity of PVPA is improved by compositing with PMVTri • The water resistance of PVPA is improved by compositing with PMVTri • The oxidative stability is greatly improved - Abstract: The poly(vinylphosphonic acid) (PVPA), poly(4-(α-methyl)vinyl-1H-1,2,3-triazole) (PMVTri), and poly(VPA-co-MVTri) were synthesized, and proton exchange membranes were prepared based on the acid-base polymeric composite of PVPA and PMVTri, and acid-base amphoteric copolymer of poly(VPA-co-MVTri). The overall performances of PVPA, proton conductivity, thermal and oxidative stability, and water resistance, are greatly improved by compositing of PMVTri or copolymerization with 4-(α-methyl)vinyl-1H-1,2,3-triazole (MVTri). About four or eight folds improvement in maximum proton conductivity was observed in the polymeric composite of PVPA/PMVTri or acid-base amphoteric copolymer poly(VPA-co-MVTri) because of the redistribution of ions in the heterostructures of PVPA and PMVTri, respectively, compared with the pristine PVPA. At the same time, the oxidative stability and the water resistance of PVPA were also greatly improved attributing to the absent of α-H in the main chain of PMVTri and the acid-base interaction between the phosphonic acid groups and the triazolyl groups, respectively

Hibernation impact on the catalytic activities of the mitochondrial D-3-hydroxybutyrate dehydrogenase in liver and brain tissues of jerboa (Jaculus orientalis

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Hafiani Assia

2003-09-01

Full Text Available Abstract Background Jerboa (Jaculus orientalis is a deep hibernating rodent native to subdesert highlands. During hibernation, a high level of ketone bodies i.e. acetoacetate (AcAc and D-3-hydroxybutyrate (BOH are produced in liver, which are used in brain as energetic fuel. These compounds are bioconverted by mitochondrial D-3-hydroxybutyrate dehydrogenase (BDH E.C. 1.1.1.30. Here we report, the function and the expression of BDH in terms of catalytic activities, kinetic parameters, levels of protein and mRNA in both tissues i.e brain and liver, in relation to the hibernating process. Results We found that: 1/ In euthemic jerboa the specific activity in liver is 2.4- and 6.4- fold higher than in brain, respectively for AcAc reduction and for BOH oxidation. The same differences were found in the hibernation state. 2/ In euthermic jerboa, the Michaelis constants, KM BOH and KM NAD+ are different in liver and in brain while KM AcAc, KM NADH and the dissociation constants, KD NAD+and KD NADH are similar. 3/ During prehibernating state, as compared to euthermic state, the liver BDH activity is reduced by half, while kinetic constants are strongly increased except KD NAD+. 4/ During hibernating state, BDH activity is significantly enhanced, moreover, kinetic constants (KM and KD are strongly modified as compared to the euthermic state; i.e. KD NAD+ in liver and KM AcAc in brain decrease 5 and 3 times respectively, while KD NADH in brain strongly increases up to 5.6 fold. 5/ Both protein content and mRNA level of BDH remain unchanged during the cold adaptation process. Conclusions These results cumulatively explained and are consistent with the existence of two BDH enzymatic forms in the liver and the brain. The apoenzyme would be subjected to differential conformational folding depending on the hibernation state. This regulation could be a result of either post-translational modifications and/or a modification of the mitochondrial membrane state

Electromechanical properties of indium–tin–oxide/poly(3,4-ethylenedioxythiophene): Poly(styrenesulfonate) hybrid electrodes for flexible transparent electrodes

International Nuclear Information System (INIS)

Jung, Sunghoon; Lim, Kyounga; Kang, Jae-Wook; Kim, Jong-Kuk; Oh, Se-In; Eun, Kyoungtae; Kim, Do-Geun; Choa, Sung-Hoon

2014-01-01

We investigated an indium–tin–oxide (ITO)/poly(3,4-ethylenedioxythiophene): poly(styrenesulfonate) (PEDOT:PSS) hybrid electrode as a potential flexible and transparent electrode. In particular, the mechanical integrity of an ITO/PEDOT:PSS hybrid electrode deposited onto a polyethylene terephthalate (PET) substrate was investigated via outer/inner bending, twisting, stretching, and adhesion tests. A PEDOT:PSS layer was inserted between ITO and PET substrate as a buffer layer to improve the flexibility and electrical properties. When a PEDOT:PSS layer was inserted, the sheet resistance of the 20 nm-thick ITO film decreased from 270 Ω/square to 57 Ω/square. Notably, the ITO/PEDOT:PSS hybrid electrode had a constant resistance change (ΔR/R 0 ) within an outer and inner bending radius of 3 mm. The bending fatigue test showed that the ITO/PEDOT:PSS hybrid electrode can withstand 10,000 bending cycles. Furthermore, the stretched ITO/PEDOT:PSS hybrid electrode showed a fairly constant resistance change up to 4%, which is more stable than the resistance change of the ITO electrode. The ITO/PEDOT:PSS electrode also shows good adhesion strength. The superior flexibility of the ITO/PEDOT:PSS hybrid electrode is attributed to the existence of a flexible PEDOT:PSS layer. This indicates that the hybridization of an ITO and PEDOT:PSS layer is a promising electrode scheme for next-generation flexible transparent electrodes. - Highlights: • We propose a hybrid electrode for flexible electronics. • Electrode made from In 2 O 3 :SnO 2 /poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) • PEDOT:PSS as a buffer layer increases flexibility and electrical conductivity. • Hybrid electrode has a superior flexibility. • Hybrid electrode can be a promising flexible transparent electrode scheme

Cellular entry of G3.5 poly (amido amine) dendrimers by clathrin- and dynamin-dependent endocytosis promotes tight junctional opening in intestinal epithelia.

Science.gov (United States)

Goldberg, Deborah S; Ghandehari, Hamidreza; Swaan, Peter W

2010-08-01

This study investigates the mechanisms of G3.5 poly (amido amine) dendrimer cellular uptake, intracellular trafficking, transepithelial transport and tight junction modulation in Caco-2 cells in the context of oral drug delivery. Chemical inhibitors blocking clathrin-, caveolin- and dynamin-dependent endocytosis pathways were used to investigate the mechanisms of dendrimer cellular uptake and transport across Caco-2 cells using flow cytometry and confocal microscopy. Dendrimer cellular uptake was found to be dynamin-dependent and was reduced by both clathrin and caveolin endocytosis inhibitors, while transepithelial transport was only dependent on dynamin- and clathrin-mediated endocytosis. Dendrimers were quickly trafficked to the lysosomes after 15 min of incubation and showed increased endosomal accumulation at later time points, suggesting saturation of this pathway. Dendrimers were unable to open tight junctions in cell monolayers treated with dynasore, a selective inhibitor of dynamin, confirming that dendrimer internalization promotes tight junction modulation. G3.5 PAMAM dendrimers take advantage of several receptor-mediated endocytosis pathways for cellular entry in Caco-2 cells. Dendrimer internalization by dynamin-dependent mechanisms promotes tight junction opening, suggesting that dendrimers act on intracellular cytoskeletal proteins to modulate tight junctions, thus catalyzing their own transport via the paracellular route.

Chemical and Enzymatic Hydrolysis of Polyurethane/Polylactide Blends

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Joanna Brzeska

2015-01-01

Full Text Available Polyether-esterurethanes containing synthetic poly[(R,S-3-hydroxybutyrate] (R,S-PHB and polyoxytetramethylenediol in soft segments and polyesterurethanes with poly(ε-caprolactone and poly[(R,S-3-hydroxybutyrate] were blended with poly([D,L]-lactide (PLA. The products were tested in terms of their oil and water absorption. Oil sorption tests of polyether-esterurethane revealed their higher response in comparison to polyesterurethanes. Blending of polyether-esterurethanes with PLA caused the increase of oil sorption. The highest water sorption was observed for blends of polyether-esterurethane, obtained with 10% of R,S-PHB in soft segments. The samples mass of polyurethanes and their blends were almost not changed after incubation in phosphate buffer and trypsin and lipase solutions. Nevertheless the molecular weight of polymers was significantly reduced after degradation. It was especially visible in case of incubation of samples in phosphate buffer what suggested the chemical hydrolysis of polymer chains. The changes of surface of polyurethanes and their blends, after incubation in both enzymatic solutions, indicated on enzymatic degradation, which had been started despite the lack of mass lost. Polyurethanes and their blends, contained more R,S-PHB in soft segments, were degraded faster.

RKIP phosphorylation–dependent ERK1 activation stimulates adipogenic lipid accumulation in 3T3-L1 preadipocytes overexpressing LC3

Energy Technology Data Exchange (ETDEWEB)

Hahm, Jong Ryeal [Department of Internal Medicine, Gyeongsang National University School of Medicine, JinJu, 527-27 (Korea, Republic of); Institute of Health Sciences, Gyeongsang National University School of Medicine, JinJu, 527-27 (Korea, Republic of); Ahmed, Mahmoud [Department of Biochemistry and Convergence Medical Science, Gyeongsang National University School of Medicine, JinJu, 527-27 (Korea, Republic of); Institute of Health Sciences, Gyeongsang National University School of Medicine, JinJu, 527-27 (Korea, Republic of); Kim, Deok Ryong, E-mail: drkim@gnu.ac.kr [Department of Biochemistry and Convergence Medical Science, Gyeongsang National University School of Medicine, JinJu, 527-27 (Korea, Republic of); Institute of Health Sciences, Gyeongsang National University School of Medicine, JinJu, 527-27 (Korea, Republic of)

2016-09-09

3T3-L1 preadipocytes undergo adipogenesis in response to treatment with dexamethaxone, 1-methyl-3-isobutylxanthine, and insulin (DMI) through activation of several adipogenic transcription factors. Many autophagy-related proteins are also highly activated in the earlier stages of adipogenesis, and the LC3 conjugation system is required for formation of lipid droplets. Here, we investigated the effect of overexpression of green fluorescent protein (GFP)-LC3 fusion protein on adipogenesis. Overexpression of GFP-LC3 in 3T3-L1 preadipocytes using poly-L-lysine-assisted adenoviral GFP-LC3 transduction was sufficient to produce intracellular lipid droplets. Indeed, GFP-LC3 overexpression stimulated expression of some adipogenic transcription factors (e.g., C/EBPα or β, PPARγ, SREBP2). In particular, SREBP2 was highly activated in preadipocytes transfected with adenoviral GFP-LC3. Also, phosphorylation of Raf kinase inhibitory protein (RKIP) at serine 153, consequently stimulating extracellular-signal regulated kinase (ERK)1 activity, was significantly increased during adipogenesis induced by either poly-L-lysine-assisted adenoviral GFP-LC3 transduction or culture in the presence of dexamethasone, 1-methyl-3-isobutylxanthine, and insulin. Furthermore, RKIP knockdown promoted ERK1 and PPARγ activation, and significantly increased the intracellular accumulation of triacylglycerides in DMI-induced adipogenesis. In conclusion, GFP-LC3 overexpression in 3T3-L1 preadipocytes stimulates adipocyte differentiation via direct modulation of RKIP-dependent ERK1 activity. - Highlights: • Overexpression of GFP-LC3 in 3T3-L1 cells produces intracellular lipid droplets. • SREBP2 is highly activated in preadipocytes transfected with adenoviral GFP-LC3. • RKIP phosphorylation at serine 153 is significantly increased during adipogenesis. • RKIP knockdown promotes ERK1 and PPARγ activation during adipogenesis. • RKIP-dependent ERK1 activation increases triacylglycerides in

Reversible adsorption of catalase onto Fe(3+) chelated poly(AAm-GMA)-IDA cryogels.

Science.gov (United States)

Aktaş Uygun, Deniz; Uygun, Murat; Akgöl, Sinan; Denizli, Adil

2015-05-01

In this presented study, poly(acrylamide-glycidyl methacrylate) [poly(AAm-GMA)] cryogels were synthesized by cryopolymerization technique at sub-zero temperature. Prepared cryogels were then functionalized with iminodiacetic acid (IDA) and chelated with Fe(3+) ions in order produce the metal chelate affinity matrix. Synthesized cryogels were characterized with FTIR, ESEM and EDX analysis, and it was found that the cryogel had sponge like structure with interconnected pores and their pore diameter was about 200 μm. Fe(3+) chelated poly(AAm-GMA)-IDA cryogels were used for the adsorption of catalase and optimum adsorption conditions were determined by varying the medium pH, initial catalase concentration, temperature and ionic strength. Maximum catalase adsorption onto Fe(3+) chelated poly(AAm-GMA)-IDA cryogel was found to be 12.99 mg/g cryogel at 25 °C, by using pH 5.0 acetate buffer. Adsorbed catalase was removed from the cryogel by using 1.0M of NaCl solution and desorption yield was found to be 96%. Additionally, reusability profile of the Fe(3+) chelated poly(AAm-GMA)-IDA cryogel was also investigated and it was found that, adsorption capacity of the cryogels didn't decrease significantly at the end of the 40 reuses. Catalase activity studies were also tested and it was demonstrated that desorbed catalase retained 70% of its initial activity. Copyright © 2015 Elsevier B.V. All rights reserved.

In vivo lactate and beta-hydroxybutyrate editing using a pure-phase refocusing pulse train.

Science.gov (United States)

Shen, J; Novotny, E J; Rothman, D L

1998-11-01

A refocusing pulse train consisting of a semiselective refocusing pulse and a selective inversion pulse to obtain a pure-phase refocusing at the frequency of maximal excitation of the semiselective refocusing pulse is proposed and applied to in vivo lactate and beta-hydroxybutyrate editing using difference spectroscopy. It is shown, using both rotation matrix theory and phantom experiments, that the soft inversion pulse has to be halved to flank the semiselective pulse to obtain perfect refocusing and cancellation of interfering resonances. The editing method is used to obtain lactate and beta-hydroxybutyrate spectra from the occipital cortex of juvenile epilepsy patients before and after ketogenic diet treatment.

Interfacial electrostatics of poly(vinylamine hydrochloride), poly(diallyldimethylammonium chloride), poly-l-lysine, and poly-l-arginine interacting with lipid bilayers.

Science.gov (United States)

McGeachy, A C; Dalchand, N; Caudill, E R; Li, T; Doğangün, M; Olenick, L L; Chang, H; Pedersen, J A; Geiger, F M

2018-04-25

Charge densities of cationic polymers adsorbed to lipid bilayers are estimated from second harmonic generation (SHG) spectroscopy and quartz crystal microbalance with dissipation monitoring (QCM-D) measurements. The systems surveyed included poly(vinylamine hydrochloride) (PVAm), poly(diallyldimethylammonium chloride) (PDADMAC), poly-l-lysine (PLL), and poly-l-arginine (PLR), as well as polyalcohol controls. Upon accounting for the number of positive charges associated with each polyelectrolyte, the binding constants and apparent free energies of adsorption as estimated from SHG data are comparable despite differences in molecular masses and molecular structure, with ΔGads values of -61 ± 2, -58 ± 2, -57 ± 1, -52 ± 2, -52 ± 1 kJ mol-1 for PDADMAC400, PDADMAC100, PVAm, PLL, and PLR, respectively. Moreover, we find charge densities for polymer adlayers of approximately 0.3 C m-2 for poly(diallyldimethylammonium chloride) while those of poly(vinylamine) hydrochloride, poly-l-lysine, and poly-l-arginine are approximately 0.2 C m-2. Time-dependent studies indicate that polycation adsorption to supported lipid bilayers is only partially reversible for most of the polymers explored. Poly(diallyldimethylammonium chloride) does not demonstrate reversible binding even over long timescales (>8 hours).

Electrochemical Interrogation of G3-Poly(propylene thiophenoimine Dendritic Star Polymer in Phenanthrene Sensing

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Hlamulo R. Makelane

2015-09-01

Full Text Available A novel dendritic star-copolymer, generation 3 poly(propylene thiophenoimine (G3PPT-co-poly(3-hexylthiophene (P3HT star co-polymer on gold electrode (i.e., Au|G3PPT-co-P3HT was used as a sensor system for the determination of phenanthrene (PHE. The G3PPT-co-P3HT star co-polymer was synthesized via in situ electrochemical co-polymerization of generation 3 poly (propylene thiophenoimine and poly (3-hexylthiophene on gold electrode. 1HNMR spectroscopy was used to determine the regioregularity of the polymer composites, whereas Fourier transform infrared spectroscopy and scanning electron microscopy were used to study their structural and morphological properties. Au|G3PPT-co-P3HT in the absence of PHE, exhibited reversible electrochemistry attributable to the oligo (thiophene ‘pendants’ of the dendrimer. PHE produced an increase in the voltammetric signals (anodic currents due to its oxidation on the dendritic material to produce catalytic current, thereby suggesting the suitability of the Au|G3PPT-co-P3HT electrode as a PHE sensor. The electrocatalysis of PHE was made possible by the rigid and planar oligo-P3HT species (formed upon the oxidation of the oligo (thiophene pendants of the star-copolymer, which allowed the efficient capture (binding and detection (electrocatalytic oxidation of PHE molecules.

Intracellular distribution of organic anions (131I-BSP and 3H-bilirubin)

International Nuclear Information System (INIS)

Kamisaka, Kazuaki; Iida, Yoshitaka; Azegami, Nobuhisa; Oda, Hiroyuki; Maezawa, Hidenori

1981-01-01

About 2 μ Ci of 131 I-BSP were injected intravenously into normal wister rats and the distributions of the isotope were determined in subcellular fractions of rat liver by the method of De Duve et al. Approximately 33% of the total activity was localized in nuclear fraction and cell debris, 28.5% was in supernatant fraction, 16.5% in microsome, 13% in lysosome and 8% in mitochondrial fraction. The subcellular distributions of radioactivity remained unchanged for 1.5 hours. Using autoradiographic method, the intracellular distribution of 3 H-bilirubin was examined by the extracted liver, 5 min, after intravenous injection of 3 H-bilirubin. 3 H-bilirubin was localized mainly in the cytoplasm and small amounts was already distributed on the canalicular membrane. It is suggested that these small molecules are mainly transported through cytoplasm and there is no specific pathway for the hepatic intracellular transport system. (author)

Gamma-hydroxybutyrate: A drug of abuse

DEFF Research Database (Denmark)

Drasbek, Kim Ryun; Christensen, Jakob; Jensen, Kimmo

2006-01-01

γ-hydroxybutyrate (GHB) is a drug of abuse that causes euphoria, anxiolysis and hypnosis. The recent rise in the recreational intake of GHB, as well as its association with “drug rape”, has turned the attention to GHB in acute hospital settings. Acutely admitted GHB intoxicated patients may display...... available. As a basis for understanding the clinical features of GHB intoxication and abuse, we here review the pharmacological and neurophysiological knowledge about GHB, which stems from decades of clinical and basic GHB research. Also, we discuss the latest discoveries in the quest for distinct GHB...... receptors in the brain, and their possible implications for future therapies of GHB abuse....

Design, Synthesis, and in Vitro Pharmacology of New Radiolabeled γ-Hydroxybutyric Acid Analogues Including Photolabile Analogues with Irreversible Binding to the High-Affinity γ-Hydroxybutyric Acid Binding Sites

DEFF Research Database (Denmark)

Sabbatini, Paola; Wellendorph, Petrine; Høg, Signe

2010-01-01

γ-Hydroxybutyric acid (GHB) is a psychotropic compound endogenous to the brain. Despite its potential physiological significance, the complete molecular mechanisms of action remain unexplained. To facilitate the isolation and identification of the high-affinity GHB binding site, we herein report ...

Thermal Annealing Effect on Poly(3-hexylthiophene: Fullerene:Copper-Phthalocyanine Ternary Photoactive Layer

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H. Derouiche

2013-01-01

Full Text Available We have fabricated poly(3-hexylthiophene (P3HT/copper phthalocyanine (CuPc/fullerene (C60 ternary blend films. This photoactive layer is sandwiched between an indium tin oxide (ITO/poly(3,4-ethylenedioxythiophene:poly(styrene sulfonate (PEDOT/PSS photoanode and a bathocuproine (BCP/aluminium photocathode. The thin films have been characterized by atomic force microscope (AFM and ultraviolet/visible spectroscopy in order to study the influence of P3HT doping on the morphological and optical properties of the photoactive layer. We have also compared the characteristics of three different organic solar cells: ITO/PEDOT:PSS/CuPc0.5:C600.5/BCP/Al and ITO/PEDOT:PSS/P3HT0.3:CuPc0.3:C600.4/BCP/Al with and without annealing. Both structures show good photovoltaic behaviour. Indeed, the incorporation of P3HT into CuPc:C60 thin film improves all the photovoltaic characteristics. We have also seen that thermal annealing significantly improves the optical absorption ability and stabilizes the organic solar cells making it more robust to chemical degradation.