Statistical Optimization of Polyhydroxybutyrate Production by <Emphasis Type="Italic">Bacillus Pumilus</Emphasis> H9 Using Cow Dung as a Cheap Carbon Source by Response Surface Methodology

A gram positive bacterium (designated strain H9) found to be a potential polyhydroxybutyrate (biodegradable polymer) producer was isolated from the soil samples of a stress prone environment (municipal waste areas). This bacterium was identified as Bacillus pumilus H9 from its morphological, physiological and 16S rRNA gene sequence analysis. A four-factor central composite rotary design was employed to optimize the medium and to find out the interactive effects of four variables, viz. concentrations of cow dung, sucrose, peptone and pH on PHB production. Using response surface methodology, a second-order polynomial equation was obtained by multiple regression analysis and a yield of 2.47 g/L of PHB dry weight was achieved from the optimized medium at pH 7. Here, we report cow dung as a cheap carbon source for the production of PHB. Further, phbA, phbB and phbC genes were amplified by polymerase chain reaction which confirms the bacterium to be able to produce polyhydroxybutyrate.     

Comparative Response of Indigenously Developed Bacterial Consortia on Progressive Degradation of Polyhydroxybutyrate Film Composites

The global demand of bioplastics has lead to an exponential increase in their production commercially. Hence, biodegradable nature needs to be evaluated in various ecosystems viz. air, water, soil and other environmental conditions to avoid the polymeric waste accumulation in the nature. In this paper, we investigated the progressive response of two indigenously developed bacterial consortia, i.e., consortium-I (C-I: Pseudomonas sp. strain Rb10, Pseudomonas sp. strain Rb11 and Bacillus sp. strain Rb18), and consortium-II (C-II: Lysinibacillus sp. strain Rb1, Pseudomonas sp. strain Rb13 and Pseudomonas sp. strain Rb19), against biodegradation behavior of polyhydroxybutyrate (PHB) film composites, under natural soil ecosystem (in net house). The biodegraded films recovered after 6 and 9 months of incubation were analyzed through Fourier transform infrared spectroscopy and scanning electron microscopy to determine the variations in chemical and morphological parameters (before and after incubation). Noticeable changes in the bond intensity, surface morphology and conductivity were found when PHB composites were treated with C-II. These changes were drastic in case of blends in comparison to copolymer. The potential isolates not only survived, but, also, there was a significant increase in bacterial diversity during whole period of incubation. To the best of our knowledge, it is the first report which described the biodegradation potential of Lysinibacillus sp. as a part of C-II with Pseudomonas sp. against PHB film composites.     

Biodegradation of Poly-ε-caprolactones and Poly-<Emphasis Type="SmallCaps">l</Emphasis>-lactides by Fungi

This work assessed biodegradation, by Aspergillus, Fusarium, Penicillium and Parengyodontium fungi, of four samples of poly-ε-caprolactone (PCL), three samples of poly-l-lactide (PLA) and one sample of poly-d,l-lactide (DL-PLA) produced by ring-opening polymerization initiated by aluminium complexes of corresponding lactones. Mesophilic fungal strains actively biodegrading PCL (F. solani) and PLA (Parengyodontium album and A. calidoustus) were selected. The rate of degradation by the selected fungi was found to depend on the physicochemical and mechanical properties of the polymers (molecular weight, polydispersity, crystallinity). The most degradable poly-ε-caprolactone sample was shown to have the lowest molecular weight; the most biodegradable polylactide DL-PLA had the lowest crystallinity. Mass spectral analysis of biodegraded polymer residues showed PCL to be degraded more intensively than PLA. It is established that in the case of Parengyodontium album the colonization of the films of polypropylene composites with DL-PLA is observed, which will undoubtedly contribute to their further destruction under the influence of abiotic factors in the environment.     

Kinetic Study of Biopolymer (PHB) Synthesis in <Emphasis Type="Italic">Alcaligenes</Emphasis> sp. in Submerged Fermentation Process Using TEM

Poly-β-hydroxybuyrate (PHB) is a carbon—energy storage material which is accumulated as intracellular granule in variety of microorganism under nutrient starved conditions. Solid PHB is a biodegradable thermoplastic polymer and is utilizable in various ways similar to many conventional plastics. Ralstonia eutropha (Alcaligenes sp.), a gram negative bacteria accumulates PHB as insoluble granules inside the cells when nutrients other than carbon are limited. In this report effort has been made to analyze PHB granule synthesis inside Alcaligenes sp. NCIM 5085 by transmission electron microscopy and qualitative estimation of PHB was carried out by fourier transform infrared spectroscopy which provide better precision compared to other conventional techniques previously applied for PHB determination. Maximum PHB concentration of 2.20 ± 0.40 g/L and cell biomass of 3.42 ± 0.20 g/L was obtained after 48.0 h of fermentation. Leudking-Piret equation deduced mixed growth associated product formation which varies from earlier reports.     

Water Absorption Kinetics and Hygrothermal Aging of Poly(lactic acid) Containing Halloysite Nanoclay and Maleated Rubber

Poly(lactic acid)/halloysite nanoclay composites (PLA/HNC) containing maleic anhydride grafted styrene-ethylene/butylene-styrene (SEBS-g-MAH) were produced using melt compounding followed by compression molding. The effects of hygrothermal aging on the thermal properties and functional groups changes of the HNC reinforced PLA (with and without SEBS-g-MAH) at three different temperatures (i.e., 30, 40 and 50 °C) were analyzed using differential scanning calorimetry and Fourier transform infrared spectroscopy techniques. The diffusion coefficient (D) of PLA was decreased by the incorporation of HNC and SEBS-g-MAH. The activation energy of water diffusion (E _a ) of PLA/HNC/SEBS-g-MAH nanocomposites was higher than that of pure PLA. The glass transition temperature (T _g ), cold-crystallization temperature (T _cc ) and melting temperature (T _m ) of the PLA sample were shifted to lower temperature and the effect was more pronounced at 50 °C. The carbonyl index values of all PLA samples increased after immersed in 40 and 50 °C, which is due to the formation of higher amount of carboxyl groups during the hydrolysis process.     

Polyhydroxybutyrate Accumulation in Bacillus megaterium and Optimization of Process Parameters Using Response Surface Methodology

The accumulation of polyhydroxybutyrate of Bacillus megaterium is growth associated and significantly dependent on carbon sources. In the present investigation B. megaterium strain isolated from soil was studied for PHB production in fructose minimal media. The PHB production was found to be growth associated. The polymer production by the strain was found to vary from 24 to 48 % content (w/w) of the dry cell weight. Box Bohn design was used to study the interactive effect of four variables on cell growth and PHB production. The optimized medium conditions with the constrain to maximize cell growth and PHB content were glucose 4.32 g/L, Mannitol 4.52 g/L and Na succinate 3.45 g/L and PHB yield 1.38 g/L amounting to 49 % of dry cell weight which is more than 1.8 folds the basal medium. The polymer production by the strain was found to vary from 12.18 to 57.2 % content (w/w) of the dry cell weight.     

New Poly(aniline-<Emphasis Type="Italic">co</Emphasis>-<Emphasis Type="Italic">o</Emphasis>-phenylenediamine)/Kaolinite Microcomposites for Water Decontamination

Copolymers of aniline and o-phenylenediamine/kaolinite composites were synthesized by 5:1 molar ratios of the respective monomers with different percentages of nanoclay via modified in situ chemical co-polymerization. The results were verified by measuring the FT-IR and UV–vis absorption spectra for PANI-o-PDA/kaolinite composites. The thermal behaviour of the copolymer and composites was studied. PANI-o-PDA/kaolinite composites were thermally more stable than pure copolymer. Surface morphology of copolymer composites was recorded at different magnification power by SEM which revealed whitish micrometric beads distributed all over the field with particle size in the range of 0.122–0.233 μm. This work demonstrates that the PANI-o-PDA/kaolinite composites particles can be considered as potential adsorbents for hazardous and toxic metal ions of water from lake El-Manzala, Egypt. All of Cd(II), Cu(II), and Pb(II) posed dangerous health risk to the local population via fish consumption.     

Catalytic co-pyrolysis of <Emphasis Type="Italic">Pterospermum acerifolium</Emphasis> and plastic waste

The continuous increase in generation of solid wastes and gradual declining of fossil fuels necessities the development of sustainable conversion technologies. Recent studies have shown that the addition of biomass with hydrogen-rich co-reactants (plastics) altogether enhances the quality of bio-fuels using pyrolysis process. It was observed that red mud (which is produced as by-product in Bayer process) was used as a catalyst in few conversion process. In this study, pyrolysis of biomass (Pterospermum acerifolium) and waste plastic mixture with activated red-mud catalyst was investigated using thermo-gravimetric analysis. The kinetic parameters (activation energy and pre-exponential factor) of this process were determined using distributed activation energy model (DAEM). The DAEM was effectively applied to decide the activation energy (E) and pre-exponential factor (A) for each sample at various conversions during the catalytic co-pyrolysis. The biomass, plastic, biomass–plastic, and biomass–plastic–catalyst exhibited activation energies in the ranges of 78–268, 172–218, 67–307, and 202–292 kJ/mol, respectively.     

A Microscopic Study of Paper Decayed by <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> and <Emphasis Type="Italic">Paecilomyces variotii</Emphasis>

The current study is interested in evaluating the decay of cotton, Whatman and chemical pulp caused by Trichoderma harzianum and Paecilomyces variotii. The structural changes of the paper were evaluated by Infrared Spectroscopy (FTIR) and Scanning Electron Microscope (SEM). The SEM results show differences in hyphae colonization and paper decay patterns between studied species under the current study; P. variotii caused an eroded structure in the cotton (cavity forming), whereas the initial T. harzianum colonization produced rupture and erosion (soft-rot decay type II) for the three types of paper ,the gaps were elongated with sharp pointed ends, which consisted either of individual cavities or in chains. Moreover, FTIR results confirmed that there a relationship could be observed between fungal decay and crystalline cellulose content because the intensity of peaks at 1335 and 1111 cm^?1 significantly decreased due to the fungal decay. Furthermore, the intensity of O–H stretching absorption slightly decreased, and this may be attributed to hydrolysis of cellulose molecules.     

Molecular Structure and Crystallization Dynamics of Poly(3-hydroxyl alkanoate) with Medium-Chain Length Biosynthesized from <Emphasis Type="Italic">Pseudomonas putida</Emphasis>

The molecular structure of poly(3-hydroxyalkanoate) (P3HA) with medium-chain length, biosynthesized from Psuedomonas putida (P. putida), was investigated by thermal analysis, X-ray diffraction, and infrared spectrum. Cultivation in a medium of nonanoic acid and a mixed substrate of nonanoic acid and glycerin as carbon sources provided P3HA with monomer units of 7 and 9 carbons (P3HA_n) and 5, 6, 7, 8, 9, and 10 carbons (P3HA_mix). Since these P3HA has comparatively long side chains, the crystallinity was as low as about 10%. It was suggested that the hydrogen bonding plays an important role in constructing the crystal. The lamellar thickness was 1.20 nm, estimated from the melting temperature depression. This lamellar thickness corresponds to two repeating units. Crystallinity depending on time was observed by the CO stretching mode in the infrared spectrum, and then Avrami’s theory was applied to analyze the crystallization mechanism. The crystallization rate of P3HA_n was very low, on the order of a few hours. An Avrami exponent of 1.45 was estimated from the slope of the Avrami plot. This shows that the conformational arrangement is presumably promoted along the chain axis. The slow crystallization is attributed to the long side chain, which prevents aggregation of the polymer chains.     

Production and Rheological Studies of Microalgal Extracellular Biopolymer from Lactose Using the Green Alga <Emphasis Type="Italic">Neochloris oleoabundans</Emphasis>

It was discovered that a green alga strain had the capacity of producing up to 5 g/L high viscosity biopolymers from lactose under mixotrophic cultivation conditions. The weight-average molecular weight of the polymer was determined to be around 505,000 Da. The ability of this strain to utilize lactose for biopolymer production promises the potential of valuable biopolymer production from cheese whey, a waste liquor stream of the cheese industry that contains approximately 50 g/L lactose. Using Plackett-Burnman experiment design and statistical analysis, it was determined that the major parameters affecting biopolymer production were the concentrations of sodium nitrate and lactose and temperature as well. Further studies on polymer characterization, production optimization, and applications are under way.     

Evaluation of the Biosorption Characteristics of <Emphasis Type="Italic">Tremella fuciformis</Emphasis> for the Decolorization of Cationic Dye from Aqueous Solution

The present work reported on the evaluation of the methylene blue dye biosorption property of Tremella fuciformis under different experimental conditions. Batch mode experiments were carried out using different experimental parameters such as initial pH, dye concentration, biosorbent amount, contact time and temperature. Four widely used kinetic models were used to elucidate the biosorption kinetics. And the kinetic analysis illustrated that the experimental data best followed the pseudo-second-order kinetic model. Biosorption equilibrium was also investigated using four widely used isotherm models. The results indicated that the experimental equilibrium data fitted very well with Langmuir isotherm models. Thermodynamic analysis of biosorption processes was found to be feasibility, spontaneous and exothermic nature of MB biosorption. These results indicated that T. fuciformis would be a high effective and environmental friendly biosorbent for MB removal from aqueous solution.     

Reductive dechlorination of a chlorinated solvent plume in Houston,Texas

Chlorinated solvents such as tetrachloroethene (perchloroethene, PCE) and trichloroethene (TCE) have been extensively used in various industrial applications for many years. Because neither are typically consumed through their various uses, they are often released to the environment through industrial application or disposal. Once released, PCE and TCE tend to migrate downward into groundwater, where they persist. In the current case study, cheese whey was used as a groundwater amendment to facilitate the reductive dechlorination of a chlorinated solvent plume underlying an auto dealer/repair shop in Harris County, Texas. From September 2010 to January 2014, over 32,000 gallons of cheese whey were injected into the subsurface resulting in a marked reduction in oxidation–reduction potential (ORP) and nitrate concentrations, coupled with an increase in ferrous iron concentrations. Statistical trend analyses indicate the primary contaminants, PCE and TCE, as well as the daughter product cis‐1,2‐dichloroethene (cDCE), all exhibited a positive response, as evidenced by statistically decreasing trends, and/or reversal in concentration trends, subsequent to cheese whey injections. Maximum concentrations of PCE and TCE in key test wells decreased by as much as 98.97 percent and 99.17 percent, respectively. In addition, the bacterial genus Dehalococcoides, capable of complete reduction of PCE to non‐toxic ethene, was found to be more abundant in the treatment area, as compared to background concentrations. Because cheese whey is a by‐product of the cheese making process, the cost of the product is essentially limited to transport. This study demonstrates cheese whey to be an effective groundwater amendment at a cost which is orders of magnitude lower than popular industry alternatives.     

Kinetic Analysis of the Temperature Effect on Polyhydroxyalkanoate Production by <Emphasis Type="Italic">Haloferax mediterranei</Emphasis> in Synthetic Molasses Wastewater

Haloferax mediterranei is an extremely halophilic archaeon that is able to synthesize polyhydroxyalkanoate (PHA) in high salt environment with low sterility demand. In this study, a mathematical model was validated and calibrated for describing the kinetic behavior of H. mediterranei at 15, 20, 25, and 35 °C in synthetic molasses wastewater. Results showed that the production of PHA by H. mediterranei, ranging from 390 to 620 mg h^?1 L^?1, was strongly dependent on the temperature. The specific growth rate (µ _max), specific substrate utilization rate (q _max), and specific decay rate (k _d) of H. mediterranei increased with temperature following Arrhenius equation prediction. The estimated activation energy was 58.31, 25.59, and 22.38 kJ mol^?1 for the process of cell growth, substrate utilization, and cell decay of H. mediterranei, respectively. The high temperature triggered the increased PHA storage even without nitrogen limitation. Thus, working at high temperatures seems a good strategy for improving the PHA productivity of H. mediterranei.     

Influence of Short Central PEO Segment on Hydrolytic and Enzymatic Degradation of Triblock PCL Copolymers

Hydrolytic, enzymatic degradation and composting under controlled conditions of series of triblock PCL/PEO copolymers, PCEC, with central short PEO block (M _n 400 g/mol) are presented and compared with homopolymer (PCL). The PCEC copolymers, synthesized via ring-opening polymerization of ε-caprolactone, were characterized by ¹H NMR, quantitative ¹³C NMR, GPC, DSC and WAXS. The introduction of the PEO central segment (<?2 wt%) in PCL chains significantly affected thermal degradation and crystallization behavior, while the hydrophobicity was slightly reduced as confirmed by water absorption and moisture uptake experiments. Hydrolytic degradation studies in phosphate buffer after 8 weeks indicated a small weight loss, while FTIR analysis detected changes in crystallinity indexes and GPC measurements revealed bulk degradation. Enzymatic degradation tested by cell-free extracts containing Pseudomonas aeruginosa PAO1 confirmed high enzyme activity throughout the surface causing morphological changes detected by optical microscopy and AFM analysis. The changes in roughness of polymer films revealed surface erosion mechanism of enzymatic degradation. Copolymer with the highest content of PEO segment and the lowest molecular weight showed better degradation ability compared to PCL and other copolymers. Furthermore, composting of polymer films in a model compost system at 37 °C resulted in significant degradation of the all synthesized block copolymers.     

Effect of <Emphasis Type="Italic">Moringa oleifera</Emphasis> Pod Husk Fibers on the Properties of Gelatin-Based Biocomposite

The objective of this work is to study the feasibility of reinforcing polymer composites by utilizing the biofibers from the agricultural residue of Moringa oleifera pod husks (MOPH). The chemical and physical properties of the fibers were comprehensively investigated to evaluate their potential as a filler in gelatin-based films. The effect of MOPH fiber concentrations of 0, 5, 10, and 15 wt% on the water vapor permeability (WVP), and mechanical and thermal properties of the gelatin-based films was studied. By incorporation of 10 wt% of the MOPH fibers in gelatin, the highest tensile strength and Young’s modulus, and the lowest WVP properties were obtained. Scanning electron microscopy (SEM) photographs indicated good interfacial adhesion between the fibers and the gelatin matrix. TGA of the biocomposites revealed an improvement of thermal stability. Moreover, under accelerated weathering, the gelatin-MOPH-10% biocomposite degraded more slowly than the gelatin control. These results indicate that the MOPH fibers are a good reinforcing filler and may be useful for biocomposite applications.     

Effect of Varying Filler Concentration on Zinc Oxide Nanoparticle Embedded Chitosan Films as Potential Food Packaging Material

Prevailing scenario of non-biodegradable food packaging materials worldwide was the motivation for this research. More than half of the packaging materials used today are non-biodegradable and lack one or the other feature that keeps it from being an ideal food packaging material. Based on the current need of food grade packaging materials, the present study illustrates the amelioration of the properties of biodegradable chitosan films with the incorporation of zinc oxide (ZnO) nanoparticles in varying concentration. The ZnO nanoparticles (ZnONPs) used as fillers in the chitosan films were synthesized by supersaturation method. They were characterized using UV–visible spectrophotometry, X-ray diffraction and field emission scanning electron microscopy (FE-SEM). The particles were observed to be around 100–200 nm in size. The chitosan films with varying concentration of ZnONPs were synthesized and characterized using Fourier transform infrared spectroscopy and FE-SEM. The films were studied for their thermal stability, water vapor transmission rate (WVTR) and mechanical properties. The thermal stability, as determined by Thermo Gravimetric Analysis and Differential Scanning Calorimetry increased slightly with increasing percentage of embedded ZnONPs while a substantial decrease in WVTR was observed. Mechanical properties also showed improvements with 77% increment in tensile modulus and 67% increment in tensile strength. The antimicrobial activity of the films was also studied on gram positive bacterium Bacillus subtilis (B. subtilis) and gram negative bacterium Escherichia coli (E. coli) by serial dilution method. A twofold and 1.5-fold increment in the antimicrobial activity was observed for B. subtilis and E. coli, respectively, with increased ZnONPs concentration in the films from 0(w/w) to 2%(w/w). Films thus prepared can prove to be of immense potential in the near future for antimicrobial food packaging applications.     

Determination of In Situ Esterification Parameters of Citric Acid-Glycerol Based Polymers for Wood Impregnation

The development of wood treatments is of increasing industrial importance. A novel technique for improving the properties of lodgepole pine and white pine through modification of the microstructure is described. The present investigation is devoted to the synthesis and determination of in situ parameters of citric acid and glycerol based polymers for wood impregnation. This solvent free approach is environmentally friendly and achieved through an esterification condensation reaction under acidic conditions. Crude glycerol and citric acid reactants were cross-linked via a curing process at 160?°C creating a polymer with only water as the byproduct. The ester bonds and crosslinking levels were controlled using different catalysts and citric acid contents and related to the reaction time and temperature. The nature of bonding within the polymers and at the wood cell walls was determined by FT-IR analysis. The thermal properties such as glass transition temperature (T_g) were studied using TGA/DSC and the effect of citric acid content and catalyst type determined. Dimensional stability of impregnated wood samples improved above 50% for each sample with HCl and p-TSA catalysts compared to control samples. FTIR spectra were studied to show the presence of the ester linkages of the polymer in situ at the wood cell walls. Bonding between the polymer and wood macromolecules were observed by scanning electron microscopy and interpreted as evidence of chemical bonds at the wood cells. When prepared using a catalyst, the polymer was intimately incorporated into wood structure significantly improving the substrate dimensional stability. Enhanced stability makes this approach of particular interest for exterior wood products especially as a green renewable option for the wood industry.     

Central Composite Design Model to Study Swelling of GrA-cl-poly(AAm) hydrogel and Kinetic Investigation of Colloidal Suspension

In the present study, the flocculation behavior of crosslinked copolymer GrA-cl-poly(AAm) hydrogel has been studied for the removal of turbidity from waste water. Sodium borohydride has been used for the reduction of the Gum rosin acids by which it gets converted into rosin alcohols. The reduced Gum rosin alcohol was crosslinked by the use of MBA and copolymerized with acrylamide using KPS as a redox initiator. Synthesized sample was then optimized for reaction conditions like reaction time, reaction temperature and the amount of solvent, monomer concentration, initiator concentration and pH of the reaction medium in order to get maximum percentage swelling. Synthesized samples were characterized using Fourier transform infrared spectroscopy, scanning electron of microscopy and X-ray diffraction techniques. Response surface methodology (RSM) based central composite design was used to study the effect of pH of swelling medium and temperature to maximize the percentage swelling. A statistical model (ANOVA) predicted pH 7.0 and temperature 40?°C as optimum operating conditions in order to get maximu swelling. GrA-cl-poly(AAm) hydrogel was found to be pH and temperature sensitive. Kaolin has been employed as a coagulant. The flocculation efficiency of the synthesized polymer was studied as a function of polymer dose, temperature and pH of the solution. GrA-cl-poly(AAm) observed to show maximum flocculation efficiency (95%) with 70mgL^?1 polymer dose in pH 5.0 at 30?°C. The adsorption capacity of malachite green dye removal (95%) was also studied with this synthesized polymer. The results validate that GrA-cl-poly(AAm) hydrogel has significant flocculation and dye removal properties and can be employed as an effective and low-cost material for removal of impurities from waste water.     

Blending Modification of PHBV/PCL and its Biodegradation by <Emphasis Type="Italic">Pseudomonas mendocina</Emphasis>

Poly(3-hydroxybutyrate-co-hydroxyvalerate) (PHBV) is a biodegradable polymer synthesized in microorganisms. The application of PHBV is limited by certain material disadvantages. Poly(ε-caprolactone) (PCL) possesses excellent thermodynamic and mechanical properties and was used to modify PHBV in the presence of triethyl citrate (TEC) and dicumyl peroxide (DCP), which was used as plasticizer and grafting agent, respectively. The effects of PCL and additive agents on the mechanical, thermal, amphipathic and degradability behaviors of the blends were investigated. The results showed that the mechanical properties of the PHBV blends improved by PCL incorporation and improved even further after TEC and DCP addition. The addition of DCP could not induce an increase in crystallization temperature but improved the crystallization degree of the blends. The presence of hydrophilic groups in TEC leads to an apparent increases in the hydrophilicity of the PHBV blends. A PHBV/PCL blend (40/60) with TEC (20 wt.%) and DCP (0.5 wt.%) was chosen for its good mechanical properties and hydrophilicity. The chosen ratio of the blends was also shown a preferable degradation activity by biodegradation assay using Pseudomonas mendocina. The addition of TEC and DCP has no conspicuous negative effect on the biodegradation.