Films of Poly(Hydroxybutyrate) (PHB) and Copper with Antibacterial Activity.

Poly(3-hydroxybutyrate), PHB, is a hydrophobic biopolymer with good mechanical and barrier properties. However, neat PHB is a semicrystalline polymer with a relative high degree of crystallinity and poor film properties. In this work, this biopolymer was plasticized with glycerol tributyrate and functionalized with copper (II) sulfate, allowing us to obtain biodegradable antimicrobial flexible films. Films with the minimum inhibitory concentration (MIC) of copper (II) sulfate presented a higher roughness than neat PHB films. The presence of plasticizer significantly improved the copper sulfate diffusion process, which was evidenced by a greater inhibition halo for plasticized materials compared to unplasticized ones, at the same salt concentration. Plasticized PHB with 2.5% copper (II) sulfate inhibited both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomona aeruginosa) bacteria, as determined by the bacterial inhibition halo. In addition, neat PHB films and PHB containing copper (II) sulfate did not show in vitro cytotoxicity in the L-929 cell line. Thus, plasticized PHB functionalized with copper (II) sulfate can be used as biodegradable antimicrobial flexible films for different applications.

Environmentally friendly starch/alginate aerogels for copper adsorption from aqueous media. A microstructural and kinetic study.

This work investigated the synthesis and characterization of alginate/starch porous materials and their application as copper ions adsorbents from aqueous media. Initially, pregel aqueous solutions with different biopolymer concentrations (1, 3, and 5% w/w) and alginate contents (25, 50, and 75% w/w) were prepared. Hydrogel formation was performed by internal and external gelation methods. Finally, the drying step was done via CO2sc leading to aerogels and via freeze-drying leading to cryogels. Process parameters influence on the final properties of materials was evaluated by BET isotherms, SEM, EDS, and TGA. Regardless the gelation method applied, interesting materials with meso- and macro-pore structure were prepared from pregel mixtures with 3% w/w biopolymer concentration and an alginate content of only 25% w/w. Low alginate content reduces the final cost of the materials. Concerning copper removal, the adsorption data were well fitted to the pseudo-second order kinetic model for aerogels and cryogels, showing aerogels the highest adsorption capacity (40 mg/g) and removal efficiency (∼ 92%). Materials demonstrated excellent reusability throughout five consecutive adsorption/desorption cycles. Hence, environmentally friendly materials with a high practical value as low-cost bioadsorbents were synthesized, having great performance in the removal of copper ions from aqueous solution.

Vinasse: from a residue to a high added value biopolymer.

This work aimed to study the feasibility of using vinasse for polyhydroxybutyrate (PHB) production by Bacillus megaterium. To optimize the culture medium, a Box-Behnken design was employed considering carbon (C), nitrogen (N), and phosphorus (Ph) concentrations as independent variables and PHB productivity as the response variable. The productivity decreased when C or N were increased, probably due to the presence of phenolic compounds and the limitation of N for the production of PHB by Bacillus sp. bacteria. An additional experimental design to optimize the C/N ratio and growing conditions (fermentation time and temperature) was carried out. Fermentation time had a statistically significant effect on PHB productivity reaching 10.6 mg/L h. On the other hand, the variability in physicochemical properties of vinasse samples led to significant differences in PHB productivity. Lower productivity values were obtained when vinasse had higher values of DBO. Therefore, biopolymers production from vinasse is a feasible alternative to valorize this bioethanol by-product.

Design and optimization of poly(hydroxyalkanoate)s production plants using alternative substrates.

In this work, we propose a Mixed Integer Nonlinear Programming (MINLP) model to determine the optimal design of a poly(hydroxyalkanoate)s (PHAs) production plant configuration. The superstructure based optimization model considers different carbon sources as raw material: glycerol (crude and purified), corn starch, cassava starch, sugarcane sucrose and sugarcane molasses. The PHA extraction section includes four alternatives: the use of enzyme, solvent, surfactant-NaOCl or surfactant-chelate. Model constraints include detailed capital cost for equipment, mass and energy balances, product specifications and operating bounds on process units. The resulting MINLP model maximizes the project net present value (NPV) as objective function and it is implemented in an equation oriented environment. Optimization results show the sugarcane-enzyme option as the most promising alternative (NPV = 75.01 million USD) for PHAs production with an energy consumption of 22.56 MJ/kg PHA and a production cost of 3.02 USD/kg PHA. Furthermore, an economic sensitivity analysis is performed.

Crystalline morphology of thermoplastic starch/talc nanocomposites induced by thermal processing.

A structural study about the changes induced by plasticization of native corn starch was carried out in this work. The influence of talc nanoparticles presence during starch thermal processing was also evaluated. Macroscopic observation of the granules appearance evolution during melt-mixing and thermo-compression was supported by a theoretical description related to these processing methods. Melt-mixing induced a polymorphic transformation from A- to Vh-type and a reduction in the degree of crystallinity. Homogenous appearance of the plasticized starch was in accordance to the disruption of granules integrity, evidenced by SEM. This observation agreed to the distinctive XRD pattern of plasticized starch from unprocessed granules. Talc incorporation did not require the adjustment of processing parameters in order to obtain a homogenous thermoplastic material, with an adequate particles distribution within the matrix. Regardless talc presence, plasticized starch presented a Vh-type crystalline structure. Thermo-compression led to particles alignment promoted by talc laminar morphology.

Modeling the bioconversion of starch to P(HB-co-HV) optimized by experimental design using Bacillus megaterium BBST4 strain.

Poly(hydroxybutyrate-co-hydroxyvalerate) (P(HB-co-HV)) is a prominent biopolymer as a potential candidate for use in the biomedical area. Several Bacillus spp. strains show promising characteristics in the use of several carbon sources and are an interesting alternative for the production of P(HB-co-HV). Sewage from the agricultural and food processing industries can be used to obtain abundantly starch as a carbon source for PHA production. The aim of the present study was to optimize by response surface methodology and desirability, the production of PHA by a Bacillus megaterium strain using starch as the sole carbon source. Two optimal conditions were determined without sporulation and were used to perform new experiments to calibrate and validate a mechanistic model, developed to simulate the dynamics of PHA and biomass production. The developed model successfully represents the kinetics of the microorganism. Employing different characterization techniques, it was determined that the PHA produced by the strain is a copolymer composed of different HB:HV proportions. Using starch as the sole carbon source in a minimal salt medium, this work shows the first reports in the literature of: 1) a mathematical model for predicting growth kinetic and PHA production for B. megaterium strain and 2) a Bacillus spp. producing P(HB-co-HV) copolymer.

Improved intracellular PHA determinations with novel spectrophotometric quantification methodologies based on Sudan black dye.

The presence of intracellular polyhydroxyalkanoates (PHAs) is usually studied using Sudan black dye solution (SB). In a previous work it was shown that the PHA could be directly quantified using the absorbance of SB fixed by PHA granules in wet cell samples. In the present paper, the optimum SB amount and the optimum conditions to be used for SB assays were determined following an experimental design by hybrid response surface methodology and desirability-function. In addition, a new methodology was developed in which it is shown that the amount of SB fixed by PHA granules can also be determined indirectly through the absorbance of the supernatant obtained from the stained cell samples. This alternative methodology allows a faster determination of the PHA content (involving 23 and 42 min for indirect and direct determinations, respectively), and can be undertaken by means of basic laboratory equipment and reagents. The correlation between PHA content in wet cell samples and the spectra of the SB stained supernatant was determined by means of multivariate and linear regression analysis. The best calibration adjustment (R2 = 0.91, RSE: 1.56%), and the good PHA prediction obtained (RSE = 1.81%), shows that the proposed methodology constitutes a reasonably precise way for PHA content determination. Thus, this methodology could anticipate the probable results of the above mentioned direct PHA determination. Compared with the most used techniques described in the scientific literature, the combined implementation of these two methodologies seems to be one of the most economical and environmentally friendly, suitable for rapid monitoring of the intracellular PHA content.

Novel spectrophotometric technique for rapid determination of extractable PHA using Sudan black dye.

Classical techniques employed to determine the amount of extractable poly(hydroxyalkanoate)s (PHAs) from cells, are laborious and destructive. Sudan black staining is commonly used in the laboratory to investigate the presence of intracellular PHA. The aim of the present study was to develop a low-cost alternative technique to achieve a quick determination of extractable intracellular PHA. This methodology employs a basic laboratory spectroscopy equipment and Sudan black dye for spectra determination. The correlation between the content of PHA in cell samples taken directly from the culture flask and its spectra was determined using partial least square regression analysis and simple linear regression analysis. The best fit obtained for calibration correlation analysis (R2=0.944, RSE: 1.24%), together with the good extractable PHA predictions (RSE=0.51%) demonstrate that the proposed methodology constitutes a fast way with high potential for the determination of extractable PHA. Based on its simplicity and flexibility, its application would be suitable in routine monitoring and rapid quantification in large-scale processes involving PHA metabolism.

Enhancement of thermoplastic starch final properties by blending with poly(ɛ-caprolactone).

Final properties of two thermoplastic corn starch matrices were improved by adding poly(ɛ-caprolactone), PCL, at 2.5, 5, and 10% w/w. One of the thermoplastic starch matrices was processed using water and glycerol as plasticizers (SG) and the other one was plasticized with a mixture of glycerol and sodium alginate (SGA). Blends were suitably processed by melt mixing and further injected. Films obtained by thermo-compression were flexible and easy to handle. Microstructure studies (SEM and FTIR) revealed a nice distribution of PCL within both matrices and also a good starch-PCL compatibility, attributed to the lower polyester concentration. The crystalline character of PCL was the responsible of the increment in the degree of crystallinity of starch matrices, determined by XRD. Moreover, it was demonstrated by TGA that PCL incorporation did not affect the thermal stability of these starch-based materials. In addition, a shift of Tg values of both glycerol and starch-rich phases to lower values was determined by DSC and DMA tests, attributed to the PCL plasticizing action. Besides, PCL blocking effect to visible and UV radiations was evident by the incremented opacity and the UV-barrier capacity of the starch films. Finally, water vapor permeability and water solubility values were reduced by PCL incorporation.

Thermoplastic starch plasticized with alginate-glycerol mixtures: Melt-processing evaluation and film properties.

Corn starch melt-processing in the presence of a commonly used plasticizer mixture (water/glycerol) and a non-conventional alternative (alginate/glycerol) was evaluated. All assayed formulations were successfully processed by melt-mixing and injected in circular probes. It was determined that all samples presented a typical viscoelastic behavior, observing a decrease in storage and loss modulus with water and alginate concentration, which facilitated samples processability. Concerning to thermal stability, it was not affected neither for water nor alginate presence. From injected probes, flexible films were obtained by thermo-compression. Films with the highest assayed water content presented a sticky appearance, whereas those containing alginate were non-tacky. Plasticizing action of water and alginate was evidenced by the occurrence of homogeneous fracture surfaces, without the presence of unmelted starch granules. Besides, the shift of glass transition temperature to lower values also corroborated the plasticizing effect of both additives. In conclusion, obtained results demonstrated the well-plasticizing action of sodium alginate on starch matrix, turning this additive into a promissory alternative to replace water during melt-processing of thermoplastic corn-starch.

Dynamics of dislocations in a two-dimensional block copolymer system with hexagonal symmetry.

Block copolymer thin films have attracted considerable attention for their ability to self-assemble into nanometre-scale architectures. Recent advances in the use of block copolymer thin films as nano-lithographic masks have driven research efforts in order to have better control of long-range ordering in the plane of the film. Irrespective of the method of sample preparation, different quasi-two-dimensional systems with hexagonal symmetry unavoidably contain translational defects, called dislocations. Dislocations control the process of coarsening in the nano/meso-scales and provide one of the most important mechanisms of length-scale selection in hexagonal patterns. Although in the last decade the nonlinear dynamics of topological defects in quasi-two-dimensional systems has witnessed significant progress, still little is known about the role of external fields on the creation and annihilation mechanisms involved in the relaxation process towards equilibrium states. In this paper, the dynamics of dislocations in non-optimal hexagonal patterns is studied in the framework of the Ohta-Kawasaki model for a diblock copolymer. Measurements of the climb and glide velocities as a function of the wave vector deformation reveal the main mechanisms of relaxation associated with the motion of dislocations.

Structure of micelles formed by highly asymmetric polystyrene-b-polydimethylsiloxane and polystyrene-b-poly[5-(N,N-diethylamino)isoprene] diblock copolymers.

The internal structure of polystyrene(PS)-shell micelles having core-forming blocks consisting of polydimethylsiloxane (PDMS) or poly[5-(N,N-diethylamino)isoprene] (PAI) was determined in detail by accessing the multilevel structural organization using static and dynamic light scattering and small-angle X-ray scattering techniques. Well-defined PS-b-PDMS and PS-b-PAI diblock copolymers with molar masses in the range of 12.0k-18.2k g/mol were dispersed in cyclohexane, dimethylacetamide, or dimethylformamide. Colloidal nanoparticles exhibiting either swollen core with a large surface area per corona chain that enables the PS chains to assume a random coil conformation with gaussian statistics, or compact core and slightly stretched PS chains in the corona were obtained. Therefore, the results of this study provide an interesting alternative allowing for precise control of the core and corona properties of PS-b-PDMS and PS-b-PAI micelles in selective solvents. Admittedly, such differences in terms of micellar properties can dictate the potential of block copolymer micelles for generating thin films from preformed nano-objects, as well as the capability to function as nanoreactors in organic medium.

Influence of the extraction-purification conditions on final properties of alginates obtained from brown algae (Macrocystis pyrifera).

In this work, three methods (ethanol, HCl, and CaCl(2) routes) of sodium alginate extraction-purification from brown seaweeds (Macrocystis pyrifera) were used in order to study the influence of process conditions on final properties of the polymer. In the CaCl(2) route, was found that the precipitation step in presence of calcium ions followed by proton-exchange in acid medium clearly gives alginates with the lowest molecular weight and poor mechanical properties. It is well known that the acid treatment degrade the ether bonds on the polymeric chain. Ethanol route displayed the best performance, where the highest yield and rheological properties were attained with the lowest number of steps. Although the polymer I.1 showed a molar mass and polydispersity index (M(w)/M(n)) similar to those of commercial sample, its mechanical properties were lower. This performance is related to the higher content of guluronic acid in the commercial alginate, which promotes a more successful calcium chelation. Moreover, the employment of pH 4 in the acid pre-treatment improved the yield of the ethanol route, avoiding the ether linkage hydrolysis. Therefore, samples I.2 and I.3 displayed a higher M(w) and a narrower distribution of molecular weights than commercial sample, which gave a higher viscosity and better viscoelastic properties.

Arm retraction potential of branched polymers in the absence of dynamic dilution.

We study the stress relaxation of model polymer networks containing low contents of star shaped and linear dangling polymers. As compared with their melts, the behavior of star and dangling polymers leads to a dynamic response with unprecedented large relaxation times. By comparing data of star melts with those corresponding to stars and dangling chains residing in polymer networks, we were able to identify the effects of dynamic dilution clearly. Since in polymer networks the dynamic dilution effect is suppressed, we were able by the first time to experimentally test the validity of the potential for arm retraction proposed by Pearson and Helfand.