Microbial degradation of polyurethane, polyester polyurethanes and polyether polyurethanes

Polyurethane (PUR) is a polymer derived from the condensation of polyisocyanate and polyol and it is widely used as a base material in various industries. PUR, in particular, polyester PUR, is known to be vulnerable to microbial attack. Recently, environmental pollution by plastic wastes has become a serious issue and polyester PUR had attracted attention because of its biodegradability. There are many reports on the degradation of polyester PUR by microorganisms, especially by fungi. Microbial degradation of polyester PUR is thought to be mainly due to the hydrolysis of ester bonds by esterases. Recently, polyester-PUR-degrading enzymes have been purified and their characteristics reported. Among them, a solid-polyester-PUR-degrading enzyme (PUR esterase) derived from Comamonas acidovorans TB-35 had unique characteristics. This enzyme has a hydrophobic PUR-surface-binding domain and a catalytic domain, and the surface-binding domain was considered as being essential for PUR degradation. This hydrophobic surface-binding domain is also observed in other solid-polyester-degrading enzymes such as poly(hydroxyalkanoate) (PHA) depolymerases. There was no significant homology between the amino acid sequence of PUR esterase and that of PHA depolymerases, except in the hydrophobic surface-binding region. Thus, PUR esterase and PHA depolymerase are probably different in terms of their evolutionary origin and it is possible that PUR esterases come to be classified as a new solid-polyester-degrading enzyme family. Received: 20 July 1998 / Received revision: 9 October 1998 / Accepted: 11 October 1998     

一株聚氨酯降解菌的分离及其降解特性解析

利用微生物对聚氨酯 (Polyurethane,PUR) 类污染物进行生物降解是目前的研究热点之一。寻找能高效降解聚氨酯的微生物是该类研究的重要前提。文中从塑料垃圾填埋场中分离培养了1株PUR高效降解菌株P10。基于菌落形态观察和16S rDNA系统发育分析,鉴定其为短芽孢杆菌属Brevibacillus的细菌。通过PUR的模式底物水性聚氨酯 (Impranil DLN) 比浊法,确定了该菌株能在6 d内降解71.4%的Impranil DLN。此外,菌株P10能够利用商业聚氨酯海绵作为唯一碳源进行生长;通过降解条件的优化,在5% (V/V) LB作为额外碳源的辅助下实现了6 d内对50 mg PUR泡沫的降解。以上结果表明Brevibacillus sp. P10在聚氨酯废弃物的生物降解过程中具有一定的应用潜力。     

Biodegradation of Polyester Polyurethane by Endophytic Fungi

Bioremediation is an important approach to waste reduction that relies on biological processes to break down a variety of pollutants. This is made possible by the vast metabolic diversity of the microbial world. To explore this diversity for the breakdown of plastic, we screened several dozen endophytic fungi for their ability to degrade the synthetic polymer polyester polyurethane (PUR). Several organisms demonstrated the ability to efficiently degrade PUR in both solid and liquid suspensions. Particularly robust activity was observed among several isolates in the genus Pestalotiopsis, although it was not a universal feature of this genus. Two Pestalotiopsis microspora isolates were uniquely able to grow on PUR as the sole carbon source under both aerobic and anaerobic conditions. Molecular characterization of this activity suggests that a serine hydrolase is responsible for degradation of PUR. The broad distribution of activity observed and the unprecedented case of anaerobic growth using PUR as the sole carbon source suggest that endophytes are a promising source of biodiversity from which to screen for metabolic properties useful for bioremediation.     

Purification and Properties of a Polyester Polyurethane-Degrading Enzyme from Comamonas acidovorans TB-35

A polyester polyurethane (PUR)-degrading enzyme, PUR esterase, derived from Comamonas acidovorans TB-35, a bacterium that utilizes polyester PUR as the sole carbon source, was purified until it showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This enzyme was bound to the cell surface and was extracted by addition of 0.2% N,N-bis(3-d-gluconamidopropyl)deoxycholamide (deoxy-BIGCHAP). The results of gel filtration and SDS-PAGE showed that the PUR esterase was a monomer with a molecular mass of about 62,000 Da. This enzyme, which is a kind of esterase, degraded solid polyester PUR, with diethylene glycol and adipic acid released as the degradation products. The optimum pH for this enzyme was 6.5, and the optimum temperature was 45 degrees C. PUR degradation by the PUR esterase was strongly inhibited by the addition of 0.04% deoxy-BIGCHAP. On the other hand, deoxy-BIGCHAP did not inhibit the activity when p-nitrophenyl acetate, a water-soluble compound, was used as a substrate. These observations indicated that this enzyme degrades PUR in a two-step reaction: hydrophobic adsorption to the PUR surface and hydrolysis of the ester bond of PUR.     

The survival of the pentachlorophenol-degrading Rhodococcus chlorophenolicus PCP-1 and Flavobacterium sp. in natural soil

The survival of two different pentachlorophenol (PCP)-degrading bacteria were studied in natural soil. The PCP-degraders Rhodococcus chlorophenolicus and Flavobacterium sp., both able to mineralize PCP into CO₂ and chloride in axenic culture, were tested for the capacity to survive and degrade PCP in natural soil. These bacteria were immobilized on polyurethane (PUR) foam and introduced into natural peaty soil to give about 10⁹ cells g^-1 of soil (dry weight). R. chlorophenolicus induced PCP-degrading activity in soil remained detectable for 200 days whether or not a carbon source was added (distillery waste or wood chips). Electron microscopic investigation performed almost a year after inoculation, revealed the presence of R. chlorophenolicus-like cells in the PUR foam particles. PCP-degrading activity of Flavobacterium sp. declined within 60 days of burial in the soil without enhancing the PCP removal. R. chlorophenolicus degraded PCP in soil at a mean rate of 3.7 mg of PCP day^-1 kg^-1 of soil, which corresponds to ca. 5×10^-3 pg of PCP degraded per inoculated R. chlorophenolicus cell day^-1. The solvent extractable organic chlorine contents of the soil decreased stoichiometrically (>95%) with that of PCP indicating that PCP was essentially mineralized.Abbreviations ATCC American type culture collection - DSM Deutsche Sammlung für Mikroorganismen - DW distillery waste - EM electron microscopy - EOX extractable organic halogen - GC/ECD gas chromatograph/electron capture detector - GC/MS gas chromatograph/mass spectrometer - PCP pentachlorophenol - WC wood chips - d.wt. dry weight - w.wt. wet weight - d.s. dry soil - d.H₂O distilled water - PCA polychlorinated aromatics     

聚氨酯塑料降解菌G-11的筛选鉴定及其塑料降解特性

聚氨酯(polyurethane,PUR)塑料因其特殊的理化性质而被广泛应用。然而,大量废弃PUR塑料的不合理处置造成了严重的资源浪费和环境污染。利用微生物的手段实现废弃PUR塑料的高效降解和循环利用成为目前的研究热点之一,而高效降解菌是PUR塑料生物法处理的关键。本研究以垃圾填埋场PUR类废塑料样品为来源,分离到一株能够降解PUR类似物Impranil DLN的微生物,并对其PUR降解特性开展了研究。通过16S rRNA基因序列比对将该菌初步鉴定为拟无枝杆菌属(Amycolatopsis sp.),命名为G-11。PUR塑料降解实验结果表明,菌株G-11对商业化PUR塑料的减重率达到4.67%,扫描电镜(scanning electron microscope,SEM)发现塑料结构被破坏,表面出现侵蚀。接触角分析和热重分析(thermogravimetric analysis,TGA)结果发现,菌株G-11处理后的PUR塑料的亲水性增强,热稳定性下降,该结果与减重和扫描电镜结果相一致。结果表明,分离自垃圾填埋场的菌株G-11在废弃PUR类塑料生物降解方面具有一定的应用潜力。     

Influence of the dimensional characteristics of polyurethane foam on high rate anaerobic digestion of piggery manure

Summay Completely mixed anaerobic reactors containing ca 10 % of their working volume as polyurethane (PUR) foam, can digest heterogenous slurries such as pig manure in a stable way at short hydraulic retention times of the order of 6.0 to 7.5 day. The influence of pore size and pad size of the PUR foam on the process was investigated. The PUR foam with the smallest pore size investigated (i.e. 45 pores per inch) gave the best biogas yield and biogas production rate. Pad size seemed to be of minor importance, having no effect on the performance of the digesters within the range examined (1 to 3.6 cm rib length). Apparently, diffusion of fatty acids into the PUR pads appeared not to be rate limiting under the lab scale reactor conditions examined.     

Concentration of proteins by foaming

Factors influencing foam concentration of proteins are studied. Projecting practical application of the results, the possibilities for obtaining good enrichment ratio are studied. The dependencies of enrichment ratio and albumin concentration in the foam on the initial solution concentration and expansion factor are investigated. Using a method of application of pressure difference in the Plateau-Gibbs borders of the foam, stabilized by albumin and lysozyme, comparatively high enrichment ratio of the proteins is obtained. The method is applicable for any protein foams and is more effective for more stable foams. The enrichment ratio of albumin significantly depends on the parameters and properties of the foam (dispersion, expansion factor, stability, etc). and also on the initial concentration of the solution. The protein concentration in the foam and the foam dispersion depend in a different way on the initial concentration by the creation of pressure difference in the foam and the R^f/C₀ dependence shows a maximum. The latter indicates the existence of an optimum of the initial protein concentration with respect to the efficiency of the foam concentration and the foam separation of proteins from solutions.     

Ferrous iron oxidation by foam immobilized Acidithiobacillus ferrooxidans: Experiments and modeling

Ferrous iron bio‐oxidation by Acidithiobacillus ferrooxidans immobilized on polyurethane foam was investigated. Cells were immobilized on foams by placing them in a growth environment and fully bacterially activated polyurethane foams (BAPUFs) were prepared by serial subculturing in batches with partially bacterially activated foam (pBAPUFs). The dependence of foam density on cell immobilization process, the effect of pH and BAPUF loading on ferrous oxidation were studied to choose operating parameters for continuous operations. With an objective to have high cell densities both in foam and the liquid phase, pretreated foams of density 50 kg/m³ as cell support and ferrous oxidation at pH 1.5 to moderate the ferric precipitation were preferred. A novel basket‐type bioreactor for continuous ferrous iron oxidation, which features a multiple effect of stirred tank in combination with recirculation, was designed and operated. The results were compared with that of a free cell and a sheet‐type foam immobilized reactors. A fivefold increase in ferric iron productivity at 33.02 g/h/L of free volume in foam was achieved using basket‐type bioreactor when compared to a free cell continuous system. A mathematical model for ferrous iron oxidation by Acidithiobacillus ferrooxidans cells immobilized on polyurethane foam was developed with cell growth in foam accounted by an effectiveness factor. The basic parameters of simulation were estimated using the experimental data on free cell growth as well as from cell attachment to foam under nongrowing conditions. The model predicted the phase of both oxidation of ferrous in shake flasks by pBAPUFs as well as by fully activated BAPUFs for different cell loadings in foam. Model for stirred tank basket bioreactor predicted within 5% both transient and steady state of the experiments closely for the simulated dilution rates. Bio‐oxidation at high Fe²⁺ concentrations were simulated with experiments when substrate and product inhibition coefficients were factored into cell growth kinetics. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Sulphur and sulphate reduction with acetate and propionate in an aerobic process for sulphide removal

Summary Sulphide production rates of sulphur-and sulphate-reducing bacteria up to 50 mg per biomass support particle per day were observed in an aerated sulphide-removal reactor with polyurethane (PUR) foam as carrier material. The optimal pH and temperature for the sulphide-producing bacteria were 8.0 and 30°C respectively. Raschig rings and four types of cube-shaped PUR particles were tested as carrier materials. When using PUR particles, the sulphide production rate was always between 3% and 4% of the sulphide removal rate, dependent on the dimensions and pore size of the polyurethane support particles. With the Raschig rings this ratio was only 2% and for reactors in which no carrier materials were present it was even lower (0.6%). Media containing different mixtures of acetate, propionate, sulphur and sulphate inoculated under anoxic conditions with sludge from the aerated reactor showed the presence of acetate-degrading sulphur-reducing, but not of acetate-degrading sulphate-reducing, bacteria. With propionate as sole electron donor no degradation occurred in the presence of sulphur within 2 weeks, whereas sulphate-dependent propionate oxidation started after 5–6 days incubation. Bacteria were isolated and resembled Desulfuromonas acetoxidans and Desulfobulbus propionicus morphologically and physiologically.     

The foams of fermentation broths. I. Some parameters of the foaming of fermentation media

The stability of foam formed during fermentation is decisively affect ed by the nature of the nutrient media used. In froth-flotation models, (a) the foam formation time, characteristic of the tendency to foam, and (b) foam subsistence time, characteristic of the stability of foams formed, have been studied. With the utilization of these two parameters, the stability of foam from aqueous solution of several surface active components of nutrient media has been noted as a function of concentrations. Further, but, without attempting completeness, the viscosity enhancing effect of carbohydrate components, and the effect of the subsistence time of their foam, upon the stability of foam have been studied together with the correlation between “standing” time after sterilization and tendency to foam. Taking soy-bean meal as a model, the stability of foam films in function of pi I, at constant concentration, has been studied. It seems that though a proper control of the factors mentioned, nutrient media with a low tendency of foaming can be formulated.     

聚氨酯塑料的微生物降解

未被合理处置的废塑料污染已成为全球性的环境问题,探索塑料废弃物的无害化处理技术势在必行。近来,研究证实了自然界中存在可以降解塑料的微生物及酶。利用微生物或酶对废塑料进行生物处理成为可能。聚氨酯塑料(Polyurethane,PUR)是广泛应用的通用塑料之一,其废弃物量已占到所有废塑料总体积的30%。文中将PUR塑料发明应用70年来有关微生物降解的研究进行了全面综述,对PUR塑料降解真菌、细菌、降解基因与酶、降解产物及相关的生物处理技术系统等进行了总结与分析,并对实现PUR废塑料高效生物处理需解决的关键科学问题进行了展望。     

Isolation of a <Emphasis Type="Italic">Pseudomonas</Emphasis><Emphasis Type="Italic">aeruginosa</Emphasis> strain from soil that can degrade polyurethane diol

Polyurethane diol (PUR-diol), a synthetic polymer, is widely used as a modifier for water-soluble resins and emulsions in wood appliances and auto coatings. Non-biodegradability of polyurethanes (PUR) and PUR-based materials poses a threat to environment that has led scientists to isolate microbes capable of degrading PUR. However, the bio-degradation of PUR-diol has not yet been reported. In this study, we report isolation of a soil bacterium that can survive using PUR-diol as sole carbon source. PUR-diol degradation by the organism was confirmed by thin layer chromatographic analysis of the conditioned medium obtained after the growth wherein a significant reduction of PUR-diol was observed compared to non-inoculated medium. To quantify the PUR-diol degradation, a sensitive assay based on High Performance Thin Layer Chromatography has been developed that showed 32% degradation of PUR-diol by the organism in 10 days. Degradation kinetics showed the maximal depletion of PUR-diol during logarithmic growth of the organism indicating a direct relation between the growth and PUR-diol degradation. Mutagenic study and GC-MS analysis revealed that esterase activity is involved in this degradation event. The ribotyping and metabolic fingerprinting analysis showed that this organism is a strain of Pseudomonous aeruginosa (P. aeruginosa). It has also been observed that this strain is able to degrade Impranil DLN™, a variety of commercially available PUR. Therefore this study identifies a new bacterium from soil that has the potential to reduce PUR-related waste burden and adds a new facet to diverse functional activities of P. aeruginosa.     

The mechanism of stabilization of actinomycete foams and the prevention of foaming under laboratory conditions

Summary Cultures ofNocardia amarae give rise to cell-stabilized foams in a laboratory scale foaming apparatus. The organism produces a surfactant and the cells are very hydrophobic; factors which, in terms of froth flotation theory, are essential for foam production and transport of the cells from the aqueous to the bubble phase. The addition of montmorillonitic clay to the culture prior to foaming prevents foam stabilization. The results obtained suggest the formation of a salt-dependent, reversible, bacterium-montmorillonite complex which prevents transport of cells to the bubble phase.     

Self-priming aerator and mechanical defoamer for microbiological processes

The design and operational characteristics of a sell-priming aerator are described. The aerator works without a compressor. H sucks the desired air quantity into the tank and distributes it, uniformly and in very fine bubbles over a certain cross-section. The design and operational characteristics of a mechanical defoamer are described. The defoamer is mounted to the top of the fermentation tank and separates foam by centrifugal force into air and liquid. It is capable of handling foams of different composition and quality without, the addition of anti foam agents. Using submerged vinegar fermentation, production of baker's yeast., and industrial waste, water treatment as examples, the performance of aerators and defoamers of different, sizes is discussed.     

Activity and batch operational stability of Candida rugosa lipase immobilized in different hydrophilic polyurethane foams during hydrolysis in a biphasic medium

The aim of this study was to investigate eventual relationships between some physico-chemical properties (e.g. porosity, aquaphilicity, partition coefficient for oleic acid and drying curves) of relatively hydrophilic polyurethane foams and the activity and batch operational stabiliy of Candida rugosa lipase immobilized within these foams. Two biocompatible polyurethane pre-polymers ("HYPOL FHP 2002^TM" and "Hypol FHP X4300^TM" from Hampshire Chemical GmbH, Germany) were tested as immobilization supports. The model reaction was the hydrolysis of crude olive residue oil in a biphasic aqueous/n-hexane medium. Drying curves under normal and reduced pressures suggested that water molecules are more strongly bound to the "FHP 2002" than to "FHP X4300" foams. This is in agreement with the higher aquaphilicity value estimated for the "FHP 2002" foam (3.7 vs 2.8). For every enzyme loading tested, hydrolysis efficiency was considerably higher for the lipase in "FHP X4300" foam when compared to the other counterpart. However, internal mass transfer limitations seem to be more severe with "FHP X4300" foams. Operational stability was evaluated in 10 consecutive batches (1 batch = 23 hours) for both immobilized preparations. A fast deactivation was observed for both biocatalysts. However, a slightly higher operational stability was observed for the lipase in "FHP 2002" foam. For the lipase in "FHP X4300" foam, the activity decay can be explained by a dramatic lipase leakage from the foam observed along successive batches. For the lipase in "FHP 2002" foam, no significant enzyme loss was observed along the reutilizations probably due to a higher number of multi-point attachment between the lipase and its support. In fact, activity and operational stability of Candida rugosa lipase in "FHP 2002" and "FHP X4300" foams appear to be related with the strength and/or the number of covalent binding between the enzyme and the support rather than to the physico-chemical properties evaluated in this work.     

Neglected Food Bubbles: The Espresso Coffee Foam

Coffee beverage known as espresso, must be topped by a velvety thick, reddish-brown foam called crema, to be considered properly prepared and to be appreciated by connoisseurs. In spite of the relevant role played by crema as a quality marker, espresso coffee foam has not yet been the subject of detailed investigations. Only recently, some aspects of the Physics and Chemistry behind the espresso coffee foam have attracted the attention of scientists. In addition to sharing several characteristics with other food foams like beer foam, for instance, the espresso coffee foam may contain solid particles (minute coffee cell-wall fragments), it is subjected to a remarkable temperature gradient and its continuous phase is an oil in water emulsion rendering it a very complex system to be studied. Moreover, in the typical regular espresso coffee cup volume (serving) of 25–30 mL, crema represents at least 10% of the total volume, and this is a limitation in obtaining experimental data by conventional instruments. The present work is aimed at reviewing the literature on espresso coffee foam. The traditional espresso brewing method will be briefly described with emphasis on the steps particularly relevant to foam formation and stabilization. In addition to present up-dated experimental data on surface properties at solid/beverage and air/beverage interface, recent advances on the espresso foam formation mechanism, as well as on foam stability, will be critically examined. The key role played by carbon dioxide generated by roasting and the effects of low and high-molecular-weight coffee compounds in promoting/inhibiting the espresso coffee foam will be discussed and emphasized.     

Aquatic macroinvertebrate biodiversity associated with artificial agricultural drainage ditches

The formation of foams on lakes is a complex phenomenon whose origin is often hardly identifiable. Recently (2007, 2008, and 2010) foam episodes started to occur in Lake Maggiore, northern Italy. The present work aimed to verify the hypothesis of an endogenous-natural origin of these foams, driven by trophic or climatic changes. To this purpose, a long-term (2000–2013) analysis of phytoplankton biovolumes, meteorological, and hydrological data has been performed together with the chemical characterization of foams. Foams resulted of endogenous origin and likely related to phytoplankton biomass degradation. Data analysis highlighted atypical warm temperature and residual lake stratification in winter in two of the three years of foam events, coupled with exceptional Bacyllariophyceae blooms in spring. Tabellaria flocculosa mostly contributed in terms of biomass in 2007 and 2008, but not in 2010; thus overall algal biomass seemed a better predictor of the risk of foam formation. Foam events occurred from July to December, driven by atypically windy conditions, and congruently with the time needed to degrade biomass into surface-active compounds. A co-occurrence of different factors resulted essential to generate foams, and climate changes likely contribute to enhance their occurrence in Lake Maggiore.     

A real-time qPCR assay for the detection of the nifH gene of Methanobrevibacter smithii, a potential indicator of sewage pollution

Aims: To degrade ether‐type polyurethane (ether‐PUR), ether‐PUR–degrading micro‐organism was isolated. Moreover, ether‐PUR–degrading mechanisms were analysed using model compounds of ether‐PUR. Methods and Results: A fungus designated as strain PURDK2, capable of changing the configuration of ether‐PUR, has been isolated. This isolated fungus was identified as Alternaria sp. Using a scanning electron microscope, the grid structure of ether‐PUR was shown to be melted and disrupted by the fungus. The degradation of ether‐PUR by the fungus was analysed, and the ether‐PUR was degraded by the fungus by about 27·5%. To analyse the urethane‐bond degradation by the fungus, a degraded product of ethylphenylcarbamate was analysed using GC/MS. Aniline and ethanol were detected by degradation with the supernatant, indicating that the fungus secreted urethane‐bond–degrading enzyme(s). PURDK2 also degraded urea bonds when diphenylmethane‐4,4′‐dibutylurea was used as a substrate. Conclusions: The enzyme(s) from PURDK2 degraded urethane and urea bonds to convert the high molecular weight structure of ether‐PUR to small molecules; and then the fungus seems to use the small molecules as an energy source. Significance and Impact of the Study: Ether‐PUR–degrading fungus, strain PURDK2, was isolated, and the urethane‐ and urea‐bonds–degrading enzymes from strain PURDK2 could contribute to the material recycling of ether‐PUR.     

The Role of Copper in Protein Foams

Chefs have known that whipping egg white proteins (EWP) in a copper bowl will improve foam stability. The improved stability is attributed to a copper–conalbumin complex or alteration of sulfhydryl reactivity. Whey proteins bind copper and show copper-induced changes in disulfide bonds; therefore, they may also be responsive to whipping in a copper bowl. EWP and whey protein isolate (WPI) solutions were whipped in the presence of 1 mM CuSO₄ or in a copper bowl with and without sugar followed by overrun and yield stress measurements and angel food cake formation. Dilational elasticity and surface tension were also measured for WPI solutions. Whipping in a copper bowl or adding 1 mM CuSO₄ significantly improved stability of EWP foams while having no effect on WPI foams. Copper caused disulfide-linked dimer formation of β-lactoglobulin and decreased dilational elasticity and surface tension, but these modifications were insufficient to change the bulk properties of foams. The addition of 10 mM CuSO₄ to WPI solutions was sufficient to increase foam stability to levels similar to EWP; however, the more stable foams formed less stable cakes. It was concluded that the effect of whipping in a copper bowl on foam properties is mainly dependent on the specific proteins forming the foam. Paper no. FSR-07-29 of the Journal Series of the Department of Food Science, North Carolina State University, Raleigh, NC 27695-7624. Presented at the 2nd International Symposium: Delivery of Functionality in Complex Food Systems: Physically-inspired Approaches From Nanoscale to Microscale, October 8–10, 2007.