Enzymatic Degradation of Cellulose-Based Materials

The biodegradability of cellulose-based materials was compared in the standard Sturm test and by enzymatic hydrolysis. Trichoderma reesei culture filtrate, the purified enzymes endoglucanase I and II from T. reesei, and -glucosidase from Aspergillus niger were used in the experiments. The unpurified Trichoderma reesei culture filtrate was found to contain a mixture of enzymes suitable for cellulose degradation. However, when purified enzymes were used the right balance of the individual enzymes was necessary. The addition of -glucosidase enhanced the enzymatic hydrolysis of cellulose materials when both culture filtrate and purified enzymes were used. In the Sturm test the biodegradability of most of the cellulose materials exceeded 70% carbon dioxide generation, but, in contrast, the biodegradability of the highly substituted aminated cellulose and cellulose acetate was below 10%. The results concerning enzymatic hydrolysis and biodegradability were in good agreement for kraft paper, sausage casing, aminated cellulose, and cellulose acetate. However, diverging results were obtained with cotton fabric, probably as a result of its high crystallinity.     

Degradation of Cellulose Acetate-Based Materials: A Review

Cellulose acetate polymer is used to make a variety of consumer products including textiles, plastic films, and cigarette filters. A review of degradation mechanisms, and the possible approaches to diminish the environmental persistence of these materials, will clarify the current and potential degradation rates of these products after disposal. Various studies have been conducted on the biodegradability of cellulose acetate, but no review has been compiled which includes biological, chemical, and photo chemical degradation mechanisms. Cellulose acetate is prepared by acetylating cellulose, the most abundant natural polymer. Cellulose is readily biodegraded by organisms that utilize cellulase enzymes, but due to the additional acetyl groups cellulose acetate requires the presence of esterases for the first step in biodegradation. Once partial deacetylation has been accomplished either by enzymes, or by partial chemical hydrolysis, the polymer’s cellulose backbone is readily biodegraded. Cellulose acetate is photo chemically degraded by UV wavelengths shorter than 280 nm, but has limited photo degradability in sunlight due to the lack of chromophores for absorbing ultraviolet light. Photo degradability can be significantly enhanced by the addition of titanium dioxide, which is used as a whitening agent in many consumer products. Photo degradation with TiO₂ causes surface pitting, thus increasing a material’s surface area which enhances biodegradation. The combination of both photo and biodegradation allows a synergy that enhances the overall degradation rate. The physical design of a consumer product can also facilitate enhanced degradation rate, since rates are highly influenced by the exposure to environmental conditions. The patent literature contains an abundance of ideas for designing consumer products that are less persistent in the outdoors environment, and this review will include insights into enhanced degradability designs.     

The effect of media changes on the rate of cellulose solubilisation by rumen and digester derived microbial communities

The aim of this study was to investigate the effect of rumen fluid and leachate-based media on the ability of rumen and anaerobic digester derived microorganisms to degrade cellulose. The results demonstrated that rumen microorganisms are not capable of solubilising cellulose, or generating biomass, at an optimal rate when grown in leachate-based media when compared to the rates achieved when grown in rumen-based media. In contrast, the rate of biomass generation and cellulose solubilisation by digester microorganisms was not strongly affected by a change in media type. Several authors have theorised that cellulose degradation rates in anaerobic digesters can be increased by inoculation with rumen-derived microorganisms. The results from this study show that this approach is unlikely to be successful, because the rumen microorganisms would likely be unable to solubilise and out-compete native solid waste microorganisms for the cellulose in a foreign (leachate based) medium.     

Biodegradation of Polycaprolactone Powders Proposed as Reference Test Materials for International Standard of Biodegradation Evaluation Method

Polycaprolactone (PCL) powders were prepared from PCL pellets using a rotation mechanical mixer. PCL powders were separated by sieves with 60 and 120 meshes into four classes; 0–125 μm, 125–250 μm, 0–250 μm and 250–500 μm. Biodegradation tests of PCL powders and cellulose powders in an aqueous solution at 25°C were performed using the coulometer according to ISO 14851. Biodegradation tests of PCL powders and cellulose powders in controlled compost at 58°C were performed by the Mitsui Chemical Analysis and Consulting Service, Inc. according to ISO 14855-1 and by using the Microbial Oxidative Degradation Analyzer (MODA) instrument according to ISO/DIS 14855-2. PCL powders were faster biodegraded than cellulose powders. The reproducibility of biodegradation of PCL powders is excellent. Differences in the biodegradation of PCL powders with different class were not observed by the ISO 14851 and ISO/DIS 14855-2. An enzymatic degradation test of PCL powders with different class was studied using an enzyme of Amano Lipase PS. PCL with smaller particle size was faster degraded by the enzyme. PCL powders with regulated sizes from 125 μm to 250 μm are proposed as a reference material for the biodegradation test.     

Study of the Determination of the Ultimate Aerobic Biodegradability of Plastic Materials Under Controlled Composting Conditions

Several ISO standards for determining the ultimate aerobic/anaerobic biodegradability of plastic materials have been published. In particular, ISO 14855-1 is a common test method that measures evolved carbon dioxide using such methods as continuous infrared analysis, gas chromatography or titration and others (ISO 14855-1(2005.9)). This method is a small-scale test for determining the ultimate aerobic biodegradability of plastic materials, where the amounts of compost inoculum and test sample in one tenth comparing with that of ISO 14855-1. This method is well versed in ISO/DIS 14855-2 which the carbon dioxide evolved from test vessel is determined by gravimetric analysis of carbon dioxide absorbent. The focus of this study is to elucidate statistically the results of round robin test by seven countries used MODA, which were various deviations among the experiments.     

Anaerobic microbial degradation of poly (3-hydroxyalkanoates) with various terminal electron acceptors

The microbial degradation of poly (3-hydroxyalkanoates) (PHAs) under anaerobic conditions with various terminal electron acceptors was examined. Nitrate-reducing consortia were established using activated sludge, and PHAs were shown to be biodegradable under these conditions. A positive correlation between carbon dioxide production and nitrate reduction was demonstrated. Nitrous oxide accumulated as the main N-containing product of nitrate reduction. The amount of PHAs in activated sludge cultures decreased approximately 20% within 40 days of incubation. Attempts were made to establish iron- and sulfate-reducing consortia from spring water, yet it could not be demonstrated that the mixed cultures were capable of degrading PHAs. Pure cultures of iron- and sulfate-reducing bacteria could not utilize PHAs as sole carbon sources. Methanogenic environments sampled included pond sediment and rumen fluid. PHAs were fermented to methane and carbon dioxide after 10 weeks by a sediment consortium, with 43 to 57% of the substrate carbon transformed to methane. Although it could not be demonstrated that PHAs were biodegraded by a rumen fluid consortium, a facultative anaerobic bacterium, identified as aStaphylococcus sp., that could grow on PHAs was isolated from rumen fluid.     

Kinetics of biochemically driven metal precipitation in synthetic landfill leachate

This study investigates the effectiveness of using metal sulphide and carbonate precipitation mechanisms combined with a landfill‐derived mixed bacterial population. The study was conducted under controlled substrate conditions in anaerobic batch reactors. High chemical oxygen demand (COD):sulphate ratios, butyrate, propionate, and acetate were used anaerobically by bacteria for growth with associated sulphate reduction as well as sulphide and carbonate generation. Propionate and butyrate degradation occurred during sulphate reduction by sulphate‐reducing bacteria while acetate degradation was associated with methanogenesis by methanogenic bacteria. Using low COD, sulphate ratios showed limited acetate utilization, but sulphate reduction still occurred. Precipitation of Cd, Cu, Zn, Ni, and Fe sulphides occurred quickly and was completed in 15 to 30 days, while Ca, Mn, and Mg carbonates formed after 40 to 50 days and some soluble metal remained even after 120 days. The rate of metal precipitation was in the order of Cd>Cu>Zn>Ni>Fe>Mn>Mg>Ca. Bacterially mediated metal precipitation occurred slower than that recognized for chemical precipitation. These findings suggest that contaminant transport models based on chemical equilibrium metal behaviors may over‐predict metal removal by bio‐precipitation. © 2002 Wiley Periodicals, Inc.     

Biodegradation of Poly(vinyl alcohol) and Bacterial Cellulose Composites by <Emphasis Type="Italic">Aspergillus niger</Emphasis>

The ability of fungal strains to attack a composite material obtained from poly(vinyl alcohol) (PVA) and bacterial cellulose (BC) is investigated. The fungal strain tested was Aspergillus niger. This fungal strain was able to change not only the polymer surface from smoother to rougher, but also to disrupt the polymer. The degradation results were confirmed by visual observations, scanning electron microscopy (SEM) analyses, X-ray diffraction analyses and FTIR spectra of the film samples. SEM micrographs confirmed the growth of fungi on the composite film surface. The degree of microbial degradation depends on culture medium and on composition of polymeric materials, especially on PVA content. The biodegradation process is accelerated by the presence of glucose in the culture medium as an easily available carbon source.     

Enhanced hydrolysis and methane yield by applying microaeration pretreatment to the anaerobic co-digestion of brown water and food waste

Microaeration has been used conventionally for the desulphurization of biogas, and recently it was shown to be an alternative pretreatment to enhance hydrolysis of the anaerobic digestion (AD) process. Previous studies on microaeration pretreatment were limited to the study of substrates with complex organic matter, while little has been reported on its effect on substrates with higher biodegradability such as brown water and food waste. Due to the lack of consistent microaeration intensities, previous studies were not comparable and thus inconclusive in proving the effectiveness of microaeration to the overall AD process. In this study, the role of microaeration pretreatment in the anaerobic co-digestion of brown water and food waste was evaluated in batch-tests. After a 4-day pretreatment with 37.5 mL-O₂/L_R-d added to the liquid phase of the reactor, the methane production of substrates were monitored in anaerobic conditions over the next 40 days. The added oxygen was consumed fully by facultative microorganisms and a reducing environment for organic matter degradation was maintained. Other than higher COD solubilization, microaeration pretreatment led to greater VFA accumulation and the conversion of other short chain fatty acids to acetate. This could be due to enhanced activities of hydrolytic and acidogenic bacteria and the degradation of slowly biodegradable compounds under microaerobic conditions. This study also found that the nature of inoculum influenced the effects of microaeration as a 21% and 10% increase in methane yield was observed when pretreatment was applied to inoculated substrates, and substrates without inoculum, respectively.     

Comparing the Growth Characteristics of Three Bacteria Involved in Degrading Rubbers

Three bacteria isolated for degradation of rubbers were compared for their growth characteristics derived from the Bradford protein assay and turbidity (optical density, OD) measurement. Both Alcaligenes xylosoxidans T2 and Pseudomonas aeruginosa GP10 were fast-growing bacteria while Nocardia corynebacterioides S3 was a slow grower utilizing rubber as the sole source of carbon and energy, but the extent of degradation was lower by the formers than the latter. A. xylosoxidans T2, P. aeruginosa GP10 and N. corynebacterioides S3 showed a typical sigmoidal growth pattern based on binding of Coomassie Brilliant Blue G250 to bacterial proteins and spectrophotometrical measurement at 600 nm. Both assays showed similar growth characteristics for all three bacteria in this study. Degradation of rubber was more pronounced by N. corynebacterioides S3 than either A. xylosoxidans T2 or P. aeruginosa GP10 during 70 days of incubation. Our results suggest that slow-growing bacteria may play a much greater role in degrading polymeric materials than was previously believed.     

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.     

Biodegradability: An assessment of commercial polymers according to the Canadian method for anaerobic conditions

The degradation of several biodegradable polymers was measured as a result of exposure to an anaerobic medium. The polymers investigated included materials based upon polylactic acid, polylactone, and poly(hydroxy butyrate/valerate) as well as those incorporating starch-based materials. The degradation was monitored by methane and carbon dioxide evolution. In addition, the physical and chemical changes were noted as a result of exposure. These measurements included changes in mass, dimension, and molecular weight. FTIR, UV-vis, proton, and¹³C NMR spectra were also recorded prior to and after exposure. The results clearly indicated that several biological and chemical degradation processes were occurring with the biodegradable polymers studied.Paper presented at the Bio/Environmentally Degradable Polymer Society—Second National Meeting, August 19–21, 1993, Chicago, Illinois.Issued as NRCC No. 37549.     

Enhancing Performance of Sphingobacterium spiritivorum in Bioremediation Phenanthrene Contaminated Sand

Many biodegradation studies have focused on survival of isolated bacteria to increase the bacteria population and subsequently enhance the efficiency of polycyclic aromatic hydrocarbon (PAH) biodegradation. However, there is limited research on enhancing the performance of isolated bacteria through reinoculation. Thus, this study was designed to evaluate the effects of reinoculation on the performance of Sphingobacterium spiritivorum in degradation of phenanthrene contaminated sand. Experiments were performed in three different reactors. Inoculation was performed once (day 0) in reactor 1. In reactor 2, inoculation was performed twice (day 0 and day 5). The bacteria was isolated from reactor 2 and inoculated into reactor 3. The study results show reactor 3 having the highest degradation rate (13.61 mg/kg/day) and percentage removal (95.36 percent). In contrast, without reinoculation in reactor 1, 68.93 percent of phenanthrene was removed. Thus, the performance of S. spiritivorum in phenanthrene degradation can be enhanced through reinoculation. © 2014 Wiley Periodicals, Inc.     

Performance and Environmental Impact of Biodegradable Films in Agriculture: A Field Study on Protected Cultivation

The performance, the degradability in soil and the environmental impact of biodegradable starch-based soil mulching and low tunnel films were assessed by means of field and laboratory tests. The lifetime of the biodegradable mulches was 9 months and of the biodegradable low-tunnel films 6 months. The radiometric properties of the biodegradable films influenced positively the microclimate: air temperature under the biodegradable low tunnel films was 2 °C higher than under the low density polyethylene films, resulting in an up to 20% higher yield of strawberries. At the end of the cultivation period, the biodegradable mulches were broken up and buried in the field soil together with the plant residues. One year after burial, less than 4% of the initial weight of the biodegradable film was found in the soil. According to ecotoxicity tests, the kinetic luminescent bacteria test with Vibrio fischeri and the Enchytraeus albidus ISO/CD 16387 reproduction potential, there was no evidence of ecotoxicity in the soil during the biodegradation process. Furthermore, there was no change in the diversity of ammonia-oxidizing bacteria in the soil determined on the basis of the appearance of amoA gene diversity in denaturing gradient gel electrophoresis.     

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.     

Characterization of Aerobic Bacteria Involved in Degrading Polyethylene Glycol (PEG)-3400 Obtained by Plating and Enrichment Culture Techniques

Polyethylene glycol (PEG) 3400-degrading aerobic bacteria were isolated from tap water and wetland sediments and then characterized. Only one Sphingomonas strain was obtained in enrichment cultures from each inoculum source whereas a total of 15 bacterial strains were isolated on agar plates. Nine of the 15 isolates were confirmed as PEG 3400 degraders. Three of the 9 PEG 3400 degraders were Gram-negative bacteria belonging to the genus Pseudomonas and genus Sphingomonas. The remaining six isolates were Gram-positive bacteria belonging to genera Rhodococcus, Williamsia, Mycobacterium and Bacillus. PEG 3400 was quantified at 194 nm spectrophotometrically and, at the same time, the growth of two Gram-negative (isolates P1 and P7) and five Gram-positive (isolates P2, P3, P4, P5 and P6) PEG 3400-degrading bacteria were assayed in liquid media and on agar plates amended with PEG 3400, and also on Nutrient Agar plates and pure agar plates without PEG 3400 addition. No growth was observed on the pure agar plates for all the tested strains for a period of 31 days. All tested PEG 3400 degraders showed much lower viability in liquid culture than on the corresponding agar plates in the presence of PEG 3400. Two Gram-negative isolates P1 and P7 did not show significant growth advantage over the Gram-positive isolates both on the agar plates and in the liquid medium amended with PEG 3400. Our results suggest that diversity of PEG degrading bacteria is high in the environments and culturing techniques affect the successful isolation of the bacteria responsible for degradation.     

Isolation of a fungus from denitrifying activated sludge that degrades highly chlorinated dioxins

 This article reports the potential of denitrifying activated sludge to degrade highly chlorinated dioxins, especially from a (landfill) leacheate treatment plant in Japan, and the isolation from this denitrifying activated sludge of a microorganism able to degrade highly chlorinated dioxins. Using a 700-ml bioreactor, denitrifying activated sludge was cultivated under denitrifying conditions by adding 2.0 ng of a mixture of 4- to 8-chlorinated dioxins from fly ash. The dioxin contents of the sample, effluent, and medium before and after cultivation were measured by gas chromatography–mass spectrometry (GC–MS). After 7 days cultivation, about 90% of added dioxins were lost (average percentage of isomer depletion). A dioxin-degrading microorganism was isolated from the activated sludge. Lignin was added to the medium as a color indicator of aromatic compound degradation, and the lignin-decolorizing microorganisms in the denitrifying activated sludge were screened. Some strains were isolated, and one major isolated fungus, strain 622, decolorized lignin effectively. Strain 622 was identified as an Acremonium sp. from its morphological characteristics. It could decolorize lignin by 24% under paraffin-sealed anaerobic conditions. After the cultivation of strain 622 with a 2 ng/ml mixture of 4- to 8-chlorinated dioxins for 1 day, 82% (average for individual isomers) of the added 4- to 8-chlorinated dioxins had been degraded. Added octachlorodibenzo-p-dioxin (OCDD, 100 ng) was degraded under aerobic conditions after 8 h of incubation. During this process, heptachlorodibenzo-p-dioxin was produced and appeared to be a degradation product of OCDD. 1- or 2-hydroxydibenzo-p-dioxin from OCDD was also identified as the degradation product by GC–MS. These results indicated that OCDD was degraded to the nonchlorinated dibenzo-p-dioxins through dechlorination by Acremonium sp. strain 622. Received: October 12, 2001 / Accepted: March 11, 2002     

Kinetic Study of Naphthalene Biodegradation in Aerobic Slurry Phase Microcosms for the Optimisation of the Process

The research was focused on the slurry-phase biodegradation of naphthalene. The biodegradation process was optimised with preliminary experiments in slurry aerobic microcosms. From soil samples collected on a contaminated site, a Pseudomonas putida strain, called M8, capable to degrade naphthalene was selected. Microcosms were prepared with M8 strain by mixing non-contaminated soil and mineral M9 medium. Different experimental conditions were tested varying naphthalene concentration, soil:water ratio and inoculum density. The disappearance of hydrocarbon, the production of carbon dioxide, and the ratio of total heterotrophic and naphthalene-degrading bacteria were monitored at different incubation times. The kinetic equation that best fitted the disappearance of contaminant with time was determined. The results showed that the isolated strain enhanced the biodegradation rate with respect to the natural biodegradation.     

The effect of lignin and sugars to the aerobic decomposition of solid wastes

A series of experimental runs were conducted from 1995 to 1999 in Madison (WI, USA) with the goal to investigate the biodegradation process of seven (7) solid waste components and mixtures of them under near optimal aerobic conditions. It was shown that substrates with high initial lignin contents or high initial HWSM contents were observed to have relatively low and high degradation extents, respectively. Two linear equations were derived that correlate degradation extent (as indicated by the volatile solids reduction) to initial lignin and initial HWSM contents separately. The lignin equation was compared to a similar equation previously developed for anaerobic environments by Chandler et al. (Predicting methane fermentation biodegradability. In: Biotechnology and Bioengineering Symposium No. 10 (1980) New York: John Wiley & Sons). With comparison to the Chandler formula, lignin was found to be less inhibitory to the overall substrate decomposition in aerobic environments compared to anaerobic ones. Cellulose loss contributed to a higher than 50% to the overall dry mass loss for all substrates studied. In addition, the cellulose to lignin (C/L) ratio appeared to be a relatively accurate compost maturity indicator, since it reduced to a value less than 0.5 for most substrates that had reached their degradation extent.     

Preparation and Characterization of Biodegradable Blend Membranes of PBS/CA

Asymmetric membranes of cellulose acetate/poly (butylene succinate) were prepared using immersion precipitation technique. The blended membranes were characterized by contact angle, scanning electron microscopy, thermogravimetric analysis, degradation test in compost and dynamic test of raisin wastewater treatment. Results demonstrated that hydrophilicity of cellulose acetate was enhanced by addition of poly (butylene succinate) up to 50 %. Furthermore, polymeric composition affects cross sectional structure of the membranes by controlling formation of macrovoids. Addition of poly (butylene succinate) improved the membranes thermal stability and obviously their degradation in compost. The prepared membranes were able to reject the wastewater pollutants properly.