Purification and biochemical characterization of recombinant alcohol dehydrogenase from the psychrophilic bacterium <Emphasis Type="Italic">Pseudomonas</Emphasis><Emphasis Type="Italic">frederiksbergensis</Emphasis>

A gram-negative psychrophilic bacterium, with potential for biodegradation of long-chain n-alkanes was isolated from ice samples collected in Spitzbergen, Denmark. On the basis of results of biochemical and morphological tests and sequence analysis of 16S rRNA, the strain was identified as Pseudomonas frederiksbergensis. In this work, a short-chain NAD⁺-dependent alcohol dehydrogenase (alcDH) (Accession number: AAR13804) from the P. frederiksbergensis was cloned and transformed in E. coli BL21 (3DE) competent cells. The alcDH activity was highest in the crude extract of cells induced with 1.0 mM IPTG. The recombinant alcDH enzyme was purified to 93.4% homogeneity using three consecutive purification steps including ammonium sulphate, Q-Sepharose Fast Flow column and gel filtration chromatography employing Superdex 200 10/30 HR column. Enzyme enrichment and yield levels of 31.4 folds and 25.5%, respectively, were achieved. While the subunit molecular mass of the enzyme was determined on SDS-PAGE to be ~38 kDa, the aggregated native form of the enzyme had a molecular mass of ~238 kDa by gel filtration analysis. Reaction conditions optima for the recombinant alcDH were determined with propan-1-ol as the substrate. While the optimum pH was 9, the optimum temperature was 35 °C. The alcDH enzyme exhibited moderate thermal stability with half-lives of 150 min at 55 °C, 27 min at 65 °C and 8 min at 75 °C. Results for kinetic parameters indicated that the apparent K _m value for alcDH with propan-1-ol as the substrate was found to be 1.42 mM and the V _max value was 0.63 mmol mg⁻¹ min⁻¹. Experimental evidence revealed that the recombinant alcDH exhibited a wide range of substrate specificity, with higher levels of specific activity for aliphatic alcohols as compared to secondary alcohols. Taken together, the present study highlights the potential of alcDH as a member of cold-adapted enzymes in several key biotechnological applications including environmental bioremediation and biotransformations. It is envisaged that, with the ongoing screening of microorganisms and metagenomes, directed evolution approaches and the subsequent overexpression of recombinant proteins, more enzymes will be found that are suitable for bioremediation purposes.     

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.     

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.     

Gene Cloning and Characterization of a Poly(<Emphasis Type="SmallCaps">l</Emphasis>-Lactic Acid) Depolymerase from <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. Strain DS04-T

A gene encoding a poly(l-lactic acid) (PLA) depolymerase from Pseudomonas sp. strain DS04-T was cloned and overexpressed in Escherichia coli. The recombinant PLA depolymerase with a molecular weight of 19.2 kDa was purified to homogeneity. The optimum pH and temperature of the PLA depolymerase are 8.5 and 60 °C, respectively. K⁺, Ca²⁺ and Ni²⁺ enhance the enzyme activity, while Na⁺, Zn²⁺, Mg²⁺, Cu²⁺, Fe²⁺, Mn²⁺ and Co²⁺ inhibit it. The inhibition of different chemicals on the PLA depolymerase activity were examined, in which EDTA was found to have a significantly inhibitory effect. The main degradation product of the depolymerase is identified as lactic acid monomer by mass spectrometric analysis. Physicochemical properties, substrate specificity and sequence analysis indicated that PME is a new type of PLA depolymerase.     

Investigation of Biosurfactant Activity and Asphaltene Biodegradation by <Emphasis Type="Italic">Bacillus cereus</Emphasis>

In this research, a biosurfactant-producing bacterium with capability of asphaltene degradation was isolated from oil-contaminated soil samples, and identified as Bacillus cereus. This strain produced an effective biosurfactant in the presence of molasses and the surface tension was reduced to the level of 36.4 mN/m after 48 h under optimum conditions. The optimum values of carbon-to-nitrogen ratio (C:N), pH, and temperature for biosurfactant production were determined as 30:1, 7.3 and 29 °C, respectively, using response surface methodology. The maximum emulsification activity in the culture broth was 53 % after 48 h using kerosene at 25 °C. The goodness of fit of four growth kinetic models including Tessier, Contois, Logistic and Westerhoff was compared for the bacterial growth and molasses utilization of B. cereus in 5-L batch bioreactor during 120 h. Conducted kinetic study showed that biosurfactant production had a good fit with the Contois growth kinetic model (R² = 0.962) and the maximum specific growth rate (µ _max ), saturation constant (K _s ) and the yield of biomass per substrate (Y _x/s ) were determined to be 0.145 h^?1, 1.83 g/L and 0.428 g/g, respectively. The asphaltene biodegradation in flask was evaluated by FTIR analysis and quantified by a spectrophotometer. This bacterium was able to degrade up to 40 % of asphaltene as a sole carbon and energy source after 60 days at 28 °C. The resulting surface tension of 30.2 mN/m with the critical micelle concentration of 23.4 mg/L indicated good efficiency of the biosurfactant.     

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.     

Enzyme-Assisted Extraction and Pb<Superscript>2+</Superscript> Biosorption of Polysaccharide from <Emphasis Type="Italic">Cordyceps militaris</Emphasis>

The enzyme assisted extraction conditions of polysaccharide from Cordyceps militaris mycelia were firstly investigated by kinetics analysis and the optimal operating was found to be: extraction temperature 40 °C; solid-solvent ratio 1:20; extraction pH 4.0; cellulase concentration 2.0%. The polysaccharide extraction yield was 5.99% under these optimized conditions. Furthermore, a fundamental investigation of the biosorption of Pb²⁺ from aqueous solution by the C. militaris polysaccharide was performed under batch conditions. The suitable pH (5.0), polysaccharide concentration (0.20 g L^?1), initial Pb²⁺ concentration (300 mg L^?1) and contact time (40 min) were outlined to enhance Pb²⁺ biosorption from aqueous medium. The Langmuir isotherm model and pseudo first order kinetic model fitted well to the data of Pb²⁺ biosorption, suggesting the biosorption of Pb²⁺ onto C. militaris polysaccharide was monolayer biosorption and physical adsorption might be the rate-limiting step that controlled the adsorption process. FTIR analysis showed that the main functional groups of C. militaris polysaccharide involved in adsorption process were carbonyl, carboxyl, and hydroxyl groups.     

Online first publication of <Emphasis Type="Italic">Journal of material cycles and waste management</Emphasis>

Announcement

Online first publication of Journal of material cycles and waste management     

Mechanism and Characterization of Polyamide 4 Degradation by <Emphasis Type="Italic">Pseudomonas</Emphasis> sp.

We identified a biodegrading microorganism of polyamide (nylon) 4, a linear polymer of γ-aminobutyric acid (GABA). From activated sludge, the biodegrading bacteria strains of Pseudomonas sp. were isolated and identified by their taxonomic characteristics and nucleotide sequences of 16S rDNA. One strain, ND-11, was grown on a minimal medium containing polyamide 4 (PA4) as the sole carbon source. The strain produced GABA as a degradation intermediate, as identified by analyzing the NMR spectra of degraded products. The culture supernatant of strain ND-11 degraded the emulsified PA4 completely within one day. These results suggest that the ND-11 strain degraded PA4 using its extracellular enzymes to hydrolyze amide bonds.     

Preparation and Characterization of Collagens from the Skin of Northern Snakehead (<Emphasis Type="Italic">Channa argus)</Emphasis>

Channa argus, a type of snakehead fish native to China, is a popular food fish in certain Asian countries but is a known destructive invasive species in the US. In this study, the two collagens, i.e. acid-soluble collagen (ASC) and pepsin-solubilized collagen (PSC), were obtained from C. argus skin. The yield of ASC was 28.0% and that of PSC was 16.8% on the dry bases. The collagens were identified as the collagen of type I by SDS–PAGE patterns. The Tds were approximately 27.0?°C. Similar ultraviolet spectra of both collagens were observed. Fourier Transform infrared spectra indicated PSC structure had a little change due to the loss of terminal domains by pepsin digestion. The results of XRD proved that the two collagens retained their helical structures. The results suggest that the collagens isolated from C. argus can potentially be alternative sources of vertebrate collagens for use in the food and other industries.     

Enhancement of biogas production potential from <Emphasis Type="Italic">Acacia</Emphasis> leaf waste using alkaline pre-treatment and co-digestion

The objective of this research was to evaluate possibility of utilizing Acacia leaves (A. mangium and A. auriculiformis), which is an agro-industrial waste from the pulp and paper industry. The effects of alkaline pre-treatment and co-digestion with Napier grass for the enhancement of biogas production from Acacia leaf waste (ALW) were investigated. Six continuous stirred tank reactors with a working volume of 5 L were carried out at the laboratory scale. The results showed that pre-treatment of Acacia leaf waste (pretreated ALW) by soaking in 3 % NaOH for 48 h increased the biogas and methane productivity to 0.200 and 0.117 m³/kgVS_added compared to 0.098 and 0.048 m³/kgVS_added of raw ALW digestion, respectively. Meanwhile, the co-digestion of Acacia leaves with different proportions of Napier grass at ratios of 1:1–1:3 in volatile solid basis also increased the production of biogas and its productivity. The maximum gas production yields of 0.424 and 0.268 m³/kgVS_added for biogas and methane were obtained at 1:3 ratio. This finding affirms the potential of ALW and its possibility to use as biogas feedstock in both single and co-substrate with Napier grass.     

Degradation of a Terephthalate-containing Polyester by Thermophilic Actinomycetes and <Emphasis Type="Italic">Bacillus</Emphasis> Species Derived from Composts

We intended to find thermophilic degraders of terephthalate-containing Biomax^® films. Films in mesh bags were buried in composts (inside temperature: approximately 55–60 °C), resulting in the degradation of them in 2 weeks. Fluorescent microscopy of films recovered from composts showed that microorganisms gradually covered the surface of a film during composting. DGGE analysis of microorganisms on the composted film indicated the presence of Bacillus species as main species (approximately 80% of microbial flora) and actinomycetes (approximately 10–20%) as the second major flora. Isolation of Biomax^®-utilizing bacteria was focused on these two genera: two actinomycetes and one Bacillus species were isolated as pure best degraders from the composted polymer films, which were fragmented into small pieces. All the strains were thermophilic and identified, based on their 16S rDNA analyses. Degradation of polymer films was confirmed by (1) accelerated fragmentation of films in composts, compared with a control (no inoculum) and resultant decrease in molecular weights, (2) growth in a powdered Biomax^® medium, compared with a control without powdered Biomax^®, and (3) production of terephthalate in a powdered Biomax^® medium. In this way, we concluded that these bacteria were useful for degradation of thermostable Biomax^® products.     

The Properties of Poly(<Emphasis Type="SmallCaps">l</Emphasis>-Lactide) Prepared by Different Synthesis Procedure

Different synthesis methods were applied to determine optimal conditions for polymerization of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (l-lactide), in order to obtain poly(l-lactide) (PLLA). Bulk polymerizations (in vacuum sealed vessel, high pressure reactor and in microwave field) were performed with tin(II) 2-ethylhexanoate as the initiator. Synthesis in the vacuum sealed vessel was carried out at the temperature of 150 °C. To reduce the reaction time second polymerization process was carried out in the high pressure reactor at 100 °C and at the pressure of 138 kPa. The third type of rapid synthesis was done in the microwave reactor at 100 °C, using frequency of 2.45 GHz and power of 150 W at the temperature of 100 °C. The temperature in this method was controlled via infrared system for in-bulk measuring. The solution polymerization (with trifluoromethanesulfonic acid as initiator) was possible even at the temperature of 40 °C, yielding PLLA with narrow molecular weight distribution in a very short period of time (less than 6 h). The obtained polymers had the number-average molecular weights ranging from 43,000 to 178,000 g mol⁻¹ (polydispersity index ranging from 1 to 3) according to the gel permeation chromatography measurements. The polymer structure was characterized by Fourier transform infrared and NMR spectroscopy. Thermal properties of the obtained polymers were investigated using thermogravimetry and differential scanning calorimetry.     

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.     

Mechanical Properties of Layered Laminated Woven Bamboo <Emphasis Type="Italic">Gigantochloa Scortechinii</Emphasis>/Epoxy Composites

This study investigated the application of bamboo as a natural composite, in which its potential as a composite material had been examined for 2–6 layers. In precise, the woven bamboo (BW) formed the culm fiber composite with an average of 0.5 mm thickness and 5.0 mm width strip. In addition, this study looked into a specific type of bamboo species known as Gigantochloa Scortechinii (Buluh Semantan), which can be found in Malaysia. This laminated plain BW, which had been reinforced with epoxy (EP), was developed by applying the hand lay-up technique. After that, the specimens were characterized via mechanical analyses, for instance, tensile, flexural, hardness, and impact tests. As a result, the 2-layer BW had displayed rather excellent results chiefly due to the incorporation of epoxy composite, although this is exceptional hardness value.     

Biosynthesis of Poly(3-hydroxybutyrate) from Cheese Whey by <Emphasis Type="Italic">Bacillus megaterium</Emphasis> NCIM 5472

Microbial polyhydroxyalkonate such as homopolyester of poly(3-hydroxybutyrate) (PHB) was produced from cheese whey by Bacillus megaterium NCIM 5472. Due to their numerous potential industrial applications, the focus was given to competently enhance the amount of PHB produced. The amount of PHB produced from whole cheese whey, and ultrafiltered cheese whey was first compared, and after observing a rise in PHB production by using ultrafiltered cheese whey, cheese whey permeate was chosen for further analysis. The presence of PHB was then confirmed by GCMS. Since the main aim of the study was to increase the amount of PHB produced through batch fermentation, various process parameters like time, pH, C/N ratio, etc. were optimized. After optimization, it was found that B. megaterium NCIM 5472 was capable of accumulating 75.5% of PHB of its dry weight and a PHB yield of 8.29 g/L. The chemical structure of the polymer was further analyzed by using FTIR and NMR spectroscopy methods. Also, the physical and thermal properties were studied by using Differential scanning calorimetry and Thermogravimetric analysis. It was found that the polymer produced had excellent thermal stability, thus allowing the possibility to exploit its properties for industrial purposes such as adhesives, packaging materials, etc.     

Synthesis and Properties of <Emphasis Type="Italic">N</Emphasis>-Maleyl Chitosan-Cross-Linked Poly(Acrylic Acid-co-Acrylamide) Superabsorbents

A high-swelling superabsorbent was synthesized with biodegradable N-maleyl chitosan as cross-linker, acrylic acid (AA) and acrylamide (AM) as the monomers, ammonium peroxodisulfate–sodium bisulfite (NaHSO₃) as redox initiation system, by means of aqueous solution polymerization. The best reaction condition was based on the orthogonal experiment design. The optimal conditions on distilled water absorbency and on 0.9 wt% NaCl solution absorbency were monomer concentration 20 wt%, mole ratio of AA to (AA + AM) 60%, the neutralization degree of AA 40%, cross-linker concentration 2% and monomer concentration 25 wt%, mole ratio of AA to (AA + AM) 60%, neutralization degree of AA 50% and cross-linker concentration 1%, respectively. Factors influencing the water absorbency of superabsorbent also were investigated, by single factor experiment method. The absorbency of superabsorbents in distilled water and 0.9 wt% NaCl solution increased and then decreased with the increasing of monomer concentration, mole ratio of AA to (AA + AM) and degree of neutralization of AA. With the increasing of cross-linker concentration, the absorbency in distilled water increased and then decreased, but it decreased all the time in 0.9 wt% NaCl solution. In enzymatic degradation tests, the weight loss of superabsorbent was related to the content of cross-linker.     

Online First publication of <Emphasis Type="Italic">Journal of Material Cycles and Waste Management</Emphasis> now available

ANNOUNCEMENT

Online First publication of Journal of Material Cycles and Waste Management now available