Biodegradability of chitin- and chitosan-containing films in soil environment

The biodegradation of polyethylene-chitin (PE-chitin) and polyethylene-chitosan (PE-chitosan) films, containing 10% by weight chitin or chitosan, by pure microbial cultures and in a soil environment was studied. Three soil-inhabited organsims,Serratia marcescens, Pseudomonas aeruginosa, andBeauveria bassiana were able to utilize chitin and chitosan in prepared PE-chitin and PE-chitosan films after eight weeks of incubation at 25°C in a basal medium containing no source of carbon or nitrogen. In a soil environment, the biodegradation of those films was studied and compared with a commercial biodegradable film containing 6% by the weight of corn starch. In soil placed in the lab, 73.4% of the chitosan and 84.7% of the chitin in the films were degraded, while 46.5% of the starch in the commercial film was degraded after six months of incubation. In an open field, 100% of the chitin and 100% of the chitosan in the films were degraded, but only 85% of the starch in the commercial film was degraded after six months of incubation. The weight of controls, (polyethylene films), remained mainly stable during the incubation period. Both PE-chitin and PE-chitosan films degraded at a higher rate than the commercial starch-based film in a soil environment indicating the potential use of chitin-based films for the manufacturing of biodegradable packaging materials.     

Microbial Poly(L-Lactide)-Degrading Enzyme Induced by Amino Acids, Peptides, and Poly(L-Amino Acids)

Poly(L-lactide)(PLA)-degrading activities of a fungus, Tritirachium album, and two strains of actinomycetes,Lentzea waywayandensis and Amycolatopsis orientalis, were inducible by some proteins (poly-L-amino acid), peptides and amino acids. Extracellular PLA-degrading activity of the culture filtrates was detected when these strains grew in liquid basal medium containing 0.1% (w/v) of (poly-L-amino acids), peptides or amino acids as the enzyme inducer. In addition to PLA-degrading activity, succinyl-(L-alanyl-L-alanyl-L-alanine)-p-nitroanilide (Suc-(Ala)₃-pNA)-degrading activity was observed, implying that the enzymes produced were protease-type. The enzyme activities produced varied between different strains and different inducers. Silk fibroin was the best inducer for A. orientalis and that elastin was the best inducer for L. waywayandensis and T. album.     

Influence of Carbon Sources on The Production and Characterization of The Exopolysaccharide (EPS) by Bacillus sphaericus 7055 Strain

In this study, EPS produced by five Bacillus spp. strains was determined. The only one strain (B. sphaericus 7055) was selected due to its high EPS production and it was investigated by growing this strain in LB broth medium containing various carbon sources. The highest EPS production of this strain was found in medium containing fructose. However, the effect of different concentrations of fructose and molasses on EPS production by the strain was studied. The maximum EPS yield of the strain 7055 was recorded with 2.5% (w/v) fructose, also the highest EPS production was found in 2.5% (w/v) molasses. The strain 7055 was found to contain (98.6%) galactose and (1.4%) glucuronic acid in control medium whereas the composition of the strain 7055 (2.5% (w/v) fructose) was found to be (99.9%) neutral sugar and (0.1%) Glucuronic acid while the strain 7055 (2.5% (w/v) molasses) was found to contain (65.9%), neutral sugars and (34.1%) glucuronic acid.     

Improvement of Cs leaching resistance of solidified radwastes with copper ferrocyanide (CFC)-vermiculite

Cesium removal from de-ionized water, seawater, and limewater using copper ferrocyanide (CFC) and porous media including silica gel, bentonite, vermiculite, and zeolite as adsorbents were investigated; CFC was incorporated with vermiculite to prepare a compound adsorbent for improving the Cs-leaching resistance of solidified borate radwastes. It was shown that the Cs-removal efficiency by CFC, defined as the percentage of cesium removed or adsorpted from solution, was largely affected by pHs of the solutions. Good removal efficiency occurred at pHs ranging from 3 to 12 with the best from 7 to 10. Vermiculite and zeolite were shown to have better removal power than silica gel and bentonite, and vermiculite was chosen to incorporate with CFC to make compound adsorbents because of its good compatibility with CFC floc. Compound adsorbents with different CFC contents were used as additives in the solidification of radioactive borate wastes for improving the cesium leaching resistance of the solidified products. Experimental results showed that the cesium leachability index measured following the method described in ANSI/ANS 16.1 increased from 7.96 to 9.76 by adding 0.25% of a compound adsorbent containing 20% CFC and 80% vermiculite. It indicated that the compound adsorbent is very useful for improving cesium-leaching resistance of the solidified borate wastes.     

Compostability of Poly(lactide): Degradation in an Inert Solid Medium

Commercial poly(lactide) degradation was studied in an inert solid medium simulating compost conditions, with the aim to achieve a complete carbon balance of the polymer degradation. The mineralisation rate at the end of the test was compared to those obtained for poly(lactide) degradation in compost. It was shown that the mineralisation rate after 45 days of degradation was quite lower in inert solid medium than in compost but the standard deviation of data was enhanced. A protocol for both extraction and quantification of the carbon included in the different degradation by-products was proposed and the carbon balance of the polymer degradation was followed during the test with a satisfactory accuracy. The non-degraded PLA material was recovered during the test, hence the evolution of the glass transition temperature and the molecular weight was followed. A two-step degradation mechanism was highlighted in inert solid medium, showing the fundamental role of abiotic reactions for PLA degradation in compost.     

Enhanced production of poly(3-hydroxybutyrate) by filamentation-suppressed recombinantEscherichia coli in a defined medium

Fed-batch cultures of recombinantEscherichia coli strains were carried out for the production of poly(3-hydroxybutyric acid) (PHB) in a chemically defined medium. TheE. coli strains used were XL1-Blue, harboring pSYL105, a stable high-copy number plasmid containing theAlcaligenes eutrophus polyhydroxyalkanoate (PHA) genes, and XL1-Blue, harboring pSYL107, which is pSYL105 containing theE. coli ftsZ gene to suppress filamentation. With XL1-Blue(pSYL105) the final cell mass and PHB concentration obtained in 62 h were 102 and 22.5 g/L, respectively. Fed-batch culture of XL1-Blue(pSYL107) under identical conditions resulted in a final cell mass and PHB concentration of 127.5 and 48.2 g/L, respectively. The PHB contents obtained with XL1-Blue(pSYL105) and XL1-Blue(pSYL107) were 22.1 and 37.8%, respectively. Therefore, PHB was more efficiently produced in a defined medium by employing filamentation-suppressed recombinantE. coli.     

Effect of vermiculite addition on compost produced from Korean food wastes

To evaluate the effect of vermiculite addition on composting food wastes from Korean households, food wastes were composted in three small bins to which different additives were added. The following three bins were employed: in Case I, only recycled compost was composted; in Case II, food wastes with recycled compost; and in Case III, food wastes with recycled compost and vermiculite. In the experiment performed for 30 days, it was confirmed that the supplementary addition of vermiculite to the composting mixture did not significantly improve the weight loss rate and the decomposition rate of food wastes. Due to dilution through the use of inorganic vermiculite, the vermiculite addition reduced the organic matter concentration of the composting mixtures. Vermiculite addition did not raise the pH value. Weight losses of roughly 70% were observed based on calculating moisture loss as well as dry food waste loss and not considering additives, while dry food waste loss was 29.4% and 35.8% with and without the addition of vermiculite, respectively. For these experiments, the major portion of the weight loss was the loss of water. The results indicate a need to differentiate between weight loss percentages and decomposition percentages, and a need to indicate if either of these percentages includes or excludes the mass of additives.     

Degradation of Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Some Soil Aspergillus spp.

Systematic screening of 45 soil fungi for degradation polyhydroxyalkanoic acids (PHAs) has led to the selection of 6 potent Aspergillus isolates belonging to A. flavus, A. oryzae, A. parasiticus, and A. racemosus. Degradation of PHAs as determined by tube assay method revealed that these Aspergillus spp. were more efficient in degrading poly(3-hydroxybutyrate) [P(3HB)] compared to copolymer of 3-hydroxybutyric acid and 3-hydroxyvaleric acid (P3HB-co-16% 3HV). Moreover, the extent of degradation in mineral base medium was much better than those in complex organic medium. For all the Aspergillus spp. tested, maximum degradation was recorded at a temperature of 37°C with significant inhibition of growth. The optimum pH range for degradation was 6.5–7.0 with degradation being maximum at pH 6.8. The extent of polymer degradation increased with increase in substrate concentration, the optimum concentration for most of the cultures being 0.4% and 0.2% (w/v) for P(3HB) and P(3HB-co-16%3HV) respectively. Supplementation of the degradation medium with additional carbon sources exerted significant inhibitory effect on both P(3HB) and P(3HB-co-16%3HV) degradation.     

Purification and Properties of Extracellular Poly(3-hydroxybutyrate) Depolymerase Produced by <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> 202

An extracellular poly(3-hydroxybutyrate) (PHB) depolymerase produced by a thermotolerant fungal soil isolate, Aspergillus fumigatus 202, was purified and characterized. Maximum PHB depolymerase production was obtained at the end of 48 h with initial medium pH 7.0 and 45 °C in Bushnell Haas Minerals medium containing PHB as sole source of carbon. The PHB depolymerase was purified using size exclusion chromatography to a fold purification of 20.62 and 61.62% yield. SDS-PAGE and isoelectric focusing revealed the molecular weight and pI of the purified enzyme as 63,744 Da and 4.2, respectively. N-terminal amino acid sequence of purified enzyme was HAXDAYLVK. This non-glycosylated enzyme was most active at pH 9.0 and 45 °C. Purified enzyme was inactivated by N-bromosuccinimide and dithiothreitol suggesting the involvement of tryptophan residues and disulfide bonds at its active site. Nonionic detergents like Tween 20, Tween 80 and Triton X-100 inhibited the enzyme activity. Ions like Ca⁺² and Mg⁺² (5 mM) increased the enzyme activity 1.5 times. Fe⁺² effectively inhibited the enzyme activity to 88% whereas Hg⁺² completely inhibited the enzyme.     

盐酸溶液改性蛭石对气态单质汞的吸附性能

采用盐酸溶液对蛭石进行浸渍处理,考察了盐酸溶液改性蛭石对气态单质汞的吸附能力。研究结果表明：盐酸溶液改性蛭石对气态单质汞的吸附效果好于未改性蛭石,盐酸浓度为4．0mol／L时,盐酸溶液改性蛭石对气态单质汞的吸附效果较佳;进气中气态单质汞的质量浓度越高,盐酸溶液改性蛭石对气态单质汞的去除率越低;适当提高吸附温度对盐酸溶液改性蛭石吸附气态单质汞较有利;降低蛭石粒径,盐酸溶液改性蛭石对气态单质汞的去除率提高。     

Large-scale production of poly(3-hydroxyvaleric acid) by fermentation ofChromobacterium violaceum,processing, and characterization of the homopolyester

A fed-batch process was developed, which allowed biotechnological production of the homopolyester poly(3-hydroxyvaleric acid) [poly(3HV)], in a mineral salts medium containing valeric acid as carbon source and complex nutrients as supplements byChromobacterium violaceum at a 10- and 300-L fermentation scale. This process yielded up to 40 g dry cell matter per L fermentation broth, and the cells contained up to 70% (w/w) poly(3HV). Poly(3HV), which was extracted from the cells with chloroform and was precipitated from this solvent with ethanol, was processed to test bars by injection molding or by press processing and to fibers by melt spinning. The unprocessed and processed poly(3HV) material was characterized with respect to the molecular weight and with respect to thermal, rheological, and mechanical properties. It was shown that it is possible to process biodegradable poly(3HV) thermoplastically and to obtain a polymer suitable for applications with low strength requirements.     

Influences of winery–distillery waste compost stability and soil type on soil carbon dynamics in amended soils

The application of organic materials to replenish soil organic matter and improve soil structure and fertility has become a common agronomic practice. This research deals with the effects of soil amendment with winery and distillery waste composts on organic carbon (C) mineralisation in two arable soils. A sandy-loam and clay-loam soil were treated and incubated with a number organic materials obtained from the co-composting of different proportions of grape stalk, grape marc, exhausted grape marc and vinasse, with sewage sludge or animal manure. Moreover, the effect of compost stability on C mineralisation dynamics was studied by applying organic materials from different stages of the composting process. The results obtained showed that the addition of exogenous organic matter stimulated microbial growth, enhanced soil respiration and increased water-extractable C contents in both soils, particularly in the days immediately following amendment. The initial composition of the different organic materials used, especially for the mature samples, and the texture of the receiving soil did not influence significantly the C mineralisation final values, with around 11–20% of the added organic C being mineralised over the first 140 days. However, the contribution of organic amendment to the labile organic C pool, maximum rates of soil respiration, as well as the extent of initial disturbance of the soil microbiota were all found to be related to the degree of organic matter stability. Moreover, irrespective of the type and stability of the organic amendment, the mineralogical composition of the receiving soil was found to significantly influence its resilience in such systems.     

Potential Remobilization of Toxic Anions during Reduction of Arsenated and Chromated Schwertmannite by the Dissimilatory Fe(III)-Reducing Bacterium Acidiphilium cryptum JF-5

Schwertmannite, an iron(III)-oxyhydroxysulfate formed in acidic mining-impacted stream or lake waters often contaminated with toxic elements like arsenate or chromate, is able to incorporate high amounts of these oxyanions. Detoxification of the water might be achieved if precipitated arsenated or chromated schwertmannite is fixed in the sediment. However, under reduced conditions, reductive dissolution of iron oxides mediated by the activity of Fe(III)-reducing bacteria might mobilize arsenate and chromate again. In this study, the reduction of synthesized arsenated or chromated schwertmannite by the acidophilic Fe(III)-reducer Acidiphilium cryptum JF-5, isolated from an acidic mining-impacted sediment, was investigated. In TSB medium at pH 2.7 with glucose as electron donor, A. cryptum JF-5 reduced about 10% of the total Fe(III) present in pure synthetic schwertmannite but only 5% of Fe(III) present in arsenated schwertmannite. In contrast to sulfate that was released during the reductive dissolution of pure schwertmannite, arsenate was not released during the reduction of arsenated schwertmannite probably due to the high surface complexation constant of arsenate and Fe(III). In medium containing chromated schwertmannite, no Fe(II) was formed, and no glucose was consumed indicating that chromate might have been toxic to cells of A. cryptum JF-5. Both As(V) or Cr(VI) could not be utilized as electron acceptor by A. cryptum JF-5. A comparison between autoclaved (121 °C for 20 min) and non-autoclaved schwertmannite samples demonstrated that nearly 100%of the bound sulfate was released during heating, and FTIR spectra indicated a transformation of schwertmannite to goethite. This structural change was not observed with autoclaved arsenated or chromated schwertmannite. These results suggest that the mobility of arsenate and chromate is not enhanced by the activity of acidophilic Fe(III)-reducing bacteria in mining-impacted sediments. In contrast, the presence of bound arsenate and chromate seemed to stabilize schwertmannite against reductive dissolution and its further transformation to goethite that is an ongoing process in those sediments.     

Synthesis of poly(hydroxyalkanoate) from hydrolyzed linseed oil

Pseudomonas putida was grown on a mixture of long-chain fatty acids obtained by hydrolysis of linseed oil. A poly(3-hydroxyalkanoate) containing 51.2% of unsaturated monomers was obtained. A considerable percentage (13.6%) was constituted by C₁₄ and C₁₆ monomers containing three double-bonds in the side chains. The polymer showed a high tendency to crosslink when it was kept in presence of air. In the crosslinked polymer no polyunsaturated monomers could be detected.     

Fungal Degradation of the Bioplastic PHB (Poly-3-hydroxy- butyric acid)

PHB (poly-3-hydroxybutyric acid) is a thermoplastic polyester synthesized by Ralstonia eutropha and other bacteria as a form of intracellular carbon and energy storage and accumulated as inclusions in the cytoplasm of these bacteria. The degradation of PHB by fungi from samples collected from various environments was studied. PHB depolymerization was tested in vials containing a PHB-containing medium which were inoculated with isolates from the samples. The degradation activity was detected by the formation of a clear zone below and around the fungal colony. In total, 105 fungi were isolated from 15 natural habitats and 8 lichens, among which 41 strains showed PHB degradation. Most of these were deuteromycetes (fungi imperfecti) resembling species of Penicillium and Aspergillus and were isolated mostly from soils, compost, hay, and lichens. Soil-containing environments were the habitats from which the largest number of fungal PHB degraders were found. Other organisms involved in PHB degradation were observed. A total number of 31 bacterial strains out of 67 isolates showed clear zones on assay medium. Protozoa, possible PHB degraders, were also found in several samples such as pond, soil, hay, horse dung, and lichen. Lichen, a fungi and algae symbiosis, was an unexpected sample from which fungal and bacterial PHB degraders were isolated.     

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.     

Biodegradation of Poly(ε-caprolactone) (PCL) Film by <Emphasis Type="Italic">Alcaligenes faecalis</Emphasis>

The biodegradation behavior of PCL film with high molecular weight (80,000 Da) in presence of bacterium Alcaligenes faecalis and the analysis of degraded polymer film have been carried out. Thin Films of PCL were prepared by means of solution casting method and the bacterial degradation behavior was carried in basal medium, in presence of bacteria with time variation after UV treatment. It was observed that after UV treatment the degradation of polymer film was increased and the degradation rate followed a three steps degradation mechanism. The degraded polymer film was analyzed by means of Differential Scanning Calorimeter (DSC), Thermo Gravimetric Analyzer (TGA) and Fourier Transform Infrared Spectroscope (FTIR). DSC results revealed that at the initial stages of the degradation up to 15–20 days, the bacterium preferentially degrades the amorphous parts of the polymer film over the crystalline zone. Thermo gravimetric analysis highlighted the low temperature stability of degraded films with extent of degradation. FTIR results showed the chain scission mechanism of the polymer chains and also supported the preferential degradation of amorphous phase over crystalline phase in the initial stages of the degradation.     

Environmentally Friendly Methodology for Preparation of Amino Acid Containing Polyamides

A short and green method for the preparation of optically active aromatic polyamides (PAs) using tetrabutylammonium bromide (TBAB) as a molten ionic liquid is reported. Polycondensation reactions of amino acid containing diacid (2S)-5-(4-methyl-2-phthalimidylpentanoylamino)isophthalic acid with various commercially available diisocyanates in molten TBAB as a green medium or in N-methylpyrrolidone as common organic solvent with or without dibutyltin dilaurate as a catalyst under microwave irradiation were carried out. Various PAs were obtained with high yields and moderate inherent viscosities in the rang of 0.30–0.57 dL/g. The obtained polymers were characterized by FT-IR, specific rotation measurements, and representative of them by ¹H NMR and elemental analysis techniques. Thermal properties of PAs were evaluated by thermogravimetric analysis and the results showed that the 10% weight loss temperature in a nitrogen atmosphere for four representative samples were more than 258 °C, which indicates that the resulting PAs have good thermal stability as well as excellent solubility.     

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.     

Potassium Release Kinetics and Water Retention of Controlled-Release Fertilizers Based on Chitosan Hydrogels

Controlled release fertilizer (CRF) hydrogels were prepared from poly(vinyl alcohol), poly(vinyl alcohol)/chitosan and chitosan using glutaraldehyde as a crosslinker. Intermolecular interactions of the CRF hydrogels were elucidated using FTIR. Water absorbency characteristics of the CRF hydrogels were also studied. It was found that the CRF hydrogels exhibited the equilibrium swelling ratio (SR) in the range 70–300%. The water retention of soil containing the CRF hydrogels was also examined. It was found that the CRF hydrogels increased the water retention of the soil. After 30 days, soil containing the PVA-, PVA/CS- and CS-hydrogels showed the water retention capacities of 25%, 10% and 4%, respectively. While the soil without the CRF hydrogel had already given off most of the water. The release behavior of potassium from the CRF hydrogels, both in deionized water and in soil, was investigated. In soil, the potassium release mechanism from the PVA- and PVA/CS-hydrogels were non-Fickian diffusion. On the other hand, the CS hydrogel showed, n value that was close to 1.0 corresponding to case II transport. In deionized water, all the CRF hydrogels showed small values of release exponent (n < 0.5) indicating a quasi-Fickian diffusion mechanism.