Poly(lactic acid) (PLA) is a compostable bioplastic manufactured by the polymerization of lactic acid monomers derived from the fermentation of starch as a feedstock. Since its first ...commercialisation in the late 1990's, PLA production has grown annually and currently it estimated that worldwide production will reach at least 800,000 tons by 2020 with Japan and the USA the two major producers. PLA is used as a replacement to conventional petrochemical based plastics, principally as food packaging containers and films and more recently, in electronics and in the manufacture of synthetic fibres. Consequently, there has been a marked increase in PLA contamination in the environment as well as increasing amounts being diverted to commercial composting facilities. This review focuses on the development, production, stability and degradation of PLA in a range of differing environments and explores our current knowledge of the environmental and biological factors involved in PLA degradation.
Poly(lactic) acid (PLA) is a compostable biopolymer and has been commercialised for the for the manufacture of short-shelf life products. As a result, increasing amounts of PLA are entering waste ...management systems and the environment; however, the degradation mechanism is unclear. While hydrolysis of the polymer occurs abiotically at elevated temperature in the presence of water, potential catalytic role for microbes in this process is yet to be established. In this study, we examined the degradation of PLA coupons from commercial packaging at a range of temperatures (25°, 37°, 45°, 50° and 55 °C) in soil and compost and compared with the degradation rates in sterile aqueous conditions by measuring loss of tensile strength and molecular weight (Mw). In addition, in order to assess the possible influence of abiotic soluble factors in compost and soil on degradation of PLA, degradation rates in microorganism-rich compost and soil were compared with sterile compost and soil extract at 50 °C. Temperature was determined to be the key parameter in PLA degradation and degradation rates in microorganism-rich compost and soil were faster than in sterile water at temperatures 45° and 50 °C determined by tensile strength and Mw loss. Furthermore, all tensile strength was lost faster after 30 and 36 days in microorganism-rich compost and soil, respectively, than in sterile compost and soil extract, 57 and 54 days, respectively at 50 °C. Significantly more Mw, 68% and 64%, was lost in compost and soil, respectively than in compost extract, Mw, 53%; and in soil extract, 57%. Therefore, degradation rates were faster in microorganism-rich compost and soil than in sterile compost and soil extract, which contained the abiotic soluble factors of compost and soil at 50 °C. These comparative studies support a direct role for microorganisms in PLA degradation at elevated temperatures in humid environments. No change in tensile strength or Mw was observed either 25° or 37 °C after 1 year suggesting that accumulation of PLA in the environment may cause future pollution issues.
•Biodegradation of four polymers (PCL, PHB, PLA and PBS) were compared.•Higher temperature was a key variable for faster biodegradation in compost.•Polycaprolactone (PCL) showed the fastest ...degradation rate under all conditions.•Aspergillus and Thermomyces fungal strains were the most abundant on all polymers.•Thermomyces lanuginosus was associated with rapid PCL degradation at 50 °C.
Plastics are an indispensable material but also a major environmental pollutant. In contrast, biodegradable polymers have the potential to be compostable. The biodegradation of four polymers as discs, polycaprolactone (PCL), polyhydroxybutyrate (PHB), polylactic acid (PLA) and poly(1,4 butylene) succinate (PBS) was compared in soil and compost over a period of more than 10 months at 25 °C, 37 °C and 50 °C. Degradation rates varied between the polymers and incubation temperatures but PCL showed the fastest degradation rate under all conditions and was completely degraded when buried in compost and incubated at 50 °C after 91 days. Furthermore, PCL strips showed a significant reduction in tensile strength in just 2 weeks when incubated in compost >45 °C. Various fungal strains growing on the polymer surfaces were identified by sequence analysis. Aspergillus fumigatus was most commonly found at 25 °C and 37 °C, while Thermomyces lanuginosus, which was abundant at 50 °C, was associated with PCL degradation.
Polylactic acid (PLA) is a compostable biopolymer that is currently used for short shelf-life product containers and waste disposal bags. At temperatures at or above its glass transition temperature ...(55°–62 °C), PLA in the presence of water undergoes chemical hydrolysis, progressively reducing polymer molecular weight and ultimately releasing lactic acid. The role of microorganisms in this process has largely focused on bacteria, with few studies investigating potential role of fungi. In this study, PLA (96% l-isomer) coupons were buried in soil or compost and incubated at 25 °C or 50 °C for up to 8 weeks. Physical disintegration of the coupons was seen only when incubated at 50 °C in both soil and compost. The principle fungal species identified from PLA coupons at 25 °C included Mortierella sp., Doratomyces microsporus, Fusarium solani, Fennellomyces sp., Aspergillus fumigatus, Verticillium sp., Lecanicillium saksenae, Cladosporium sp. and Trichoderma sp. Only Thermomyces lanuginosus and A. fumigatus were isolated from PLA at 50 °C and both were shown to cause significant PLA biodegradation as determined by tensile strength loss compared to abiotic controls. Terminal restriction fragment length polymorphism (TRFLP) demonstrated strong selection of specific fungal species from soil and compost communities on PLA the surface and a greater species diversity than could be detected by conventional culturing techniques.
•Colonisation & biodegradation of poly(lactic) acid coupons (96% l-isomer) in soil & compost were investigated.•Fungi colonising PLA showed a high degree of selection from the surrounding community.•PLA tensile strength loss was associated with significant fungal colonisation at 50 °C.•Aspergillus fumigatus &Thermomyces lanuginosus were the principle fungi colonising PLA.•A. fumigatus and T. lanuginosus were both shown to accelerate PLA degradation in monoculture.
Abstract
Fungi are known to have an important role in the composting process as degraders of recalcitrant materials such as cellulose and lignin. Previous attempts to study the diversity and ...succession of fungi in compost systems have relied on the use of culture-dependent analyses and low-resolution DNA-fingerprinting techniques, lacking the necessary depth to analyse such a rich ecosystem. In this study, 454 pyrosequencing was used to characterize the fungal community composition at the different stages of an in-vessel composting process. A complex succession of fungi was revealed, with 251 fungal OTUs identified throughout the monitoring period. The Ascomycota were the dominant phylum (82.5% of all sequences recovered), followed by the Basidiomycota (10.4%) and the subphylum Mucoromycotina (4.9%). In the starting materials and early stages of the process, yeast species from the Saccharomycetales were abundant, while in latter stages and in the high temperature regions of the pile, fungi from the orders Eurotiales, Sordariales, Mucorales, Agaricales and Microascales were the most prominent. This study provides an improved understanding of the fungal diversity occurring during the composting of municipal solid waste, and this knowledge can lead to the development of more efficient composting practices and a better evaluation of the end-product quality.
Fungal succession in an in-vessel composting system characterized using 454 pyrosequencing.
Environmental populations of the opportunistic pathogen Aspergillus fumigatus have been shown to be genotypically diverse and to contain a range of isolates with varying pathogenic potential. In this ...study, we combined two RAPD primers to investigate the genetic diversity of environmental isolates from Manchester collected monthly over 1 year alongside Dublin environmental isolates and clinical isolates from patients. RAPD analysis revealed a diverse genotype, but with three major clinical isolate clusters. When the pathogenicity of clinical and Dublin isolates was compared with a random selection of Manchester isolates in a Galleria mellonella larvae model, as a group, clinical isolates were significantly more pathogenic than environmental isolates. Moreover, when relative pathogenicity of individual isolates was compared, clinical isolates were the most pathogenic, Dublin isolates were the least pathogenic and Manchester isolates showed a range in pathogenicity. Overall, this suggests that the environmental population is genetically diverse, displaying a range in pathogenicity, and that the most pathogenic strains from the environment are selected during patient infection.
Filamentous fungi such as Aspergillus niger have a high capacity secretory system and are therefore widely exploited for the industrial production of native and heterologous proteins. However, in ...most cases the yields of non-fungal proteins are significantly lower than those obtained for fungal proteins. One well-studied bottleneck appears to be the result of mis-folding of heterologous proteins in the ER during early stages of secretion, with related stress responses in the host, including the unfolded protein response (UPR). This study aims at uncovering transcriptional and translational responses occurring in A. niger exposed to secretion stress.
A genome-wide transcriptional analysis of protein secretion-related stress responses was determined using Affymetrix DNA GeneChips and independent verification for selected genes. Endoplasmic reticulum (ER)-associated stress was induced either by chemical treatment of the wild-type cells with dithiothreitol (DTT) or tunicamycin, or by expressing a human protein, tissue plasminogen activator (t-PA). All of these treatments triggered the UPR, as shown by the expression levels of several well-known UPR target genes. The predicted proteins encoded by most of the up-regulated genes function as part of the secretory system including chaperones, foldases, glycosylation enzymes, vesicle transport proteins, and ER-associated degradation proteins. Several genes were down-regulated under stress conditions and these included several genes that encode secreted enzymes. Moreover, translational regulation under ER stress was investigated by polysomal fractionation. This analysis confirmed the post-transcriptional control of hacA expression and highlighted that differential translation also occurs during ER stress, in particular for some genes encoding secreted proteins or proteins involved in ribosomal biogenesis and assembly.
This is first genome-wide analysis of both transcriptional and translational events following protein secretion stress. Insight has been gained into the molecular basis of protein secretion and secretion-related stress in an effective protein-secreting fungus, and provides an opportunity to identify target genes for manipulation in strain improvement strategies.
Aspergillus fumigatus, an opportunistic fungal pathogen that causes invasive aspergillosis in immunosuppressed patients, is considered to be the world's most dangerous mould. It is widely distributed ...in the environment, and airborne asexual conidia serve as the main mode of transport for pulmonary lung infection. It is important to monitor seasonal airborne conidia levels when assessing the risk of acquiring this infection. In this study, air was sampled for total viable fungal spores and viable A. fumigatus conidia monthly over a 2-year period (2009 and 2010) close to Manchester, UK, city center. Total viable airborne fungal counts varied seasonally, peaking in the summer and autumn for both years and reaching levels of approximately 1100–1400 colony-forming units (CFU)/m3; counts were strongly positively correlated to mean temperature (R
2 = 0.697). By contrast, A. fumigatus viable airborne counts were not seasonally associated; persistent low levels were between 3 and 20 CFU/m3 and were not correlated with mean temperature (R
2 = 0.018). A total of 220 isolates of A. fumigatus were recovered on potato dextrose agar (PDA) at 45°C, and internal transcribed spacer sequencing and restriction digestion of a partial polymerase chain reaction amplicon of the β-tubulin gene (benA) of 34 randomly selected isolates were used to confirm the isolates as A. fumigatus. When the colony radial growth rates (Kr) were determined, the highest rates were observed on PDA, followed by Vogel's medium supplemented with phosphatidylcholine and Vogel's medium alone. Clinical isolates had a significantly higher mean colony Kr on PDA compared with environmental isolates.
The compostable biopolymer, poly(lactic) acid (PLA), is increasingly being used as an alternative to conventional plastics for short shelf-life products, disposable bags and packaging, and in ...agriculture. Despite the increase in the amount of PLA entering composting systems, few studies have examined the impact of PLA degradation on the compost microbial community. Thermophilic fungi play an import role in the composting process as they secrete hydrolytic enzymes capable of breaking down an array of complex natural polymers. In this study, the impact of PLA hydrolysis on the compost fungal community was examined by terminal restriction fragment length polymorphism and 454 sequencing. At 25°C, the effect of PLA on the surrounding compost community was relatively small and no physical changes were observed to the PLA films. However, when incubated at 50°C, where physical disintegration of PLA was occurring, a clear divergence between the compost populations in the presence and absence of PLA was evident after 2 months but became closer to the population in the absence of PLA after 4 months indicating that, after causing an initial perturbation after 2 months, the population began to return to that seen in the absence of PLA. The only exception was in the population containing 50% (w/w) PLA film, which remained divergent after 4 months and was associated with a marked acidification of the compost. Thus, 454-pyrosequencing revealed that the presence of PLA caused a strong selection for a Thermomyces sp. that was present only at low abundance in the absence of PLA.
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