聚乳酸塑料合成、生物降解及其废弃物处置的研究进展

随着国内外禁塑令和限塑令的升级,以聚乳酸(polylactic acid, PLA)为代表的生物基塑料成为传统石油基塑料市场的主要替代品,备受产业界的青睐。然而,公众对生物基塑料的认识仍存在诸多误解。事实上,生物基塑料的降解需要在特定条件下才能实现,泄入到自然环境中同样难以降解,会对人体、生物多样性和生态系统功能造成危害,这与传统石油基塑料相似。近年来,随着我国PLA产能和市场规模不断的提高,亟需进一步加强对PLA等生物基塑料降解性能的认识,挖掘PLA生物降解资源,关注和研究生物基塑料回收处理模式。基于上述背景,本文首先介绍了PLA塑料的性质及合成方式,以及PLA塑料的产业化与市场规模;其次,对目前聚乳酸塑料微生物与酶法降解的研究进展进行了综述,并对其生物降解机制进行了探讨;最后,提出了微生物原位处理和酶法闭环回收两种聚乳酸塑料废弃物生物处置方法,并对PLA生物基塑料的发展前景和趋势进行了展望。     

聚乳酸(PLA)生物降解的研究进展

聚乳酸(Polylactic Acid,PLA)是一种新兴的,由可再生资源--乳酸聚合而成的高分子聚酯.因为其具有优良的物理化学性能、生物相容性及生物可降解性,且对环境及人体无毒害作用,而被认为是一种最具潜力的绿色生物塑料.作为环境友好材料,聚乳酸日益受到人们的重视.基于可循环利用的考虑,其生物降解的研究也成为当前研究的一个重要方面.本文综述了PLA生物降解领域的相关进展,包括降解的微生物学、相关酶学及分子生物学,系统阐述了PLA可能的生物降解机制.并对生物系统处理PLA废弃物的可行性进行了探讨.     

聚对苯二甲酸乙二醇酯生物降解的研究进展

塑料广泛存在于人类的日常生活中，在给人们生活带来便利的同时，大量塑料废物也给环境带来很大压力。聚对苯二甲酸乙二醇酯(polyethylene terephthalate, PET)是一种以石油为原料的高分子热塑性材料，因其具有耐用、透明度高、重量轻等特性，已成为世界上使用最广泛的塑料之一。由于PET具有结构复杂以及难降解的特性，可在自然界中长期存在，不仅对全球生态环境造成严重的污染，而且已经威胁到人类健康。如何对PET废弃物进行降解已成为全球的难题之一，相较于物理法和化学法，生物降解法是目前处理PET废弃物最为绿色环保的方法。本文分别介绍了微生物和生物酶对PET生物降解的研究现状、PET的生物降解途径、PET生物降解机制以及PET降解酶的分子改造等方面的研究，并对如何实现PET的高效降解、寻找和改造可降解高结晶度PET的微生物或酶进行展望，为PET的生物降解微生物或酶的有效开发应用提供理论依据。     

The enzymatic hydrolysis of starch-based PVOH and polyol plasticised blends

Thermoplastic starch (TPS) materials present several advantages to the plastic industry and when blended with other materials they can exhibit improved mechanical and moisture sensitivity properties compared to pure TPS materials. However, the biodegradability of these blends, through such processes as enzymatic degradation, needs to be characterised to ensure the beneficial properties of TPS are not compromised. The aims of the study were to investigate the effect of varying polyvinyl alcohol (PVOH) content and polyol type within the TPS blends on the rate and extent of starch enzymatic hydrolysis using enzymes α-amylase and amyloglucosidase. The results of this study have revealed that TPS:PVOH blends with a PVOH content at 50 wt% exhibited a significantly reduced rate and extent of starch hydrolysis. The results suggest that this may have been attributed to interactions between starch and PVOH that further prevented enzymatic attack on the remaining starch phases within the blend. The extent of starch hydrolysis was not significantly affected by polyol type, however, the rate of starch hydrolysis from the maltitol blend was significantly reduced compared to sorbitol and glycerol substrates.     

Micromechanical Modeling Study of Mechanical Inhibition of Enzymatic Degradation of Collagen Tissues

This study investigates how the collagen fiber structure influences the enzymatic degradation of collagen tissues. We developed a micromechanical model of a fibrous collagen tissue undergoing enzymatic degradation based on two central hypotheses. The collagen fibers are crimped in the undeformed configuration. Enzymatic degradation is an energy activated process and the activation energy is increased by the axial strain energy density of the fiber. We determined the intrinsic degradation rate and characteristic energy for mechanical inhibition from fibril-level degradation experiments and applied the parameters to predict the effect of the crimped fiber structure and fiber properties on the degradation of bovine cornea and pericardium tissues under controlled tension. We then applied the model to examine the effect of the tissue stress state on the rate of tissue degradation and the anisotropic fiber structures that developed from enzymatic degradation.     

Membrane composition and successful bioaugmentation. Studies of the interactions of model thylakoid and plasma cyanobacterial and bacterial membranes with fungal membrane-lytic enzyme Lecitase ultra

Cyanobacterial/bacterial consortia are frequently inoculated to soils to increase the soil fertility and to accelerate the biodegradation of organic pollutants. Moreover, such consortia can also be successfully applied in landfills especially for the biodegradation of plastic wastes. However, the bioaugmentation techniques turn out frequently inefficient due to the competition of the indigenous microorganisms attacking directly these inoculated or secreting to their surroundings cell wall and membrane-lytic enzymes. It can be hypothesized that the resistance of the microbial membrane to the enzymatic degradation is correlated with its lipid composition. To verify this hypothesis glycolipid and phospholipid Langmuir monolayers were applied as models of thylakoid and plasma cyanobacterial and bacterial membranes. Hybrid fungal enzyme Lecitase ultra joining the activity of lipase and phospholipase A1 was applied as the model of fungal membrane-lytic enzyme. It turned out that anionic thylakoid lipids sulfoquinovosyldiacylglycerol and phosphatidylglycerols were the main targets of Lecitase ultra in the model multicomponent thylakoid membranes. The resistance of the model plasma bacterial membranes to enzymatic degradation depended significantly to their composition. The resistance increased generally when the unsaturated lipids were exchanged to their saturated counterparts. However, most resistant turned out the membranes composed of unsaturated phosphatidylamine and saturated anionic phospholipids.     

Biofilm and Diatom Succession on Polyethylene (PE) and Biodegradable Plastic Bags in Two Marine Habitats: Early Signs of Degradation in the Pelagic and Benthic Zone?

The production of biodegradable plastic is increasing. Given the augmented littering of these products an increasing input into the sea is expected. Previous laboratory experiments have shown that degradation of plastic starts within days to weeks. Little is known about the early composition and activity of biofilms found on biodegradable and conventional plastic debris and its correlation to degradation in the marine environment. In this study we investigated the early formation of biofilms on plastic shopper bags and its consequences for the degradation of plastic. Samples of polyethylene and biodegradable plastic were tested in the Mediterranean Sea for 15 and 33 days. The samples were distributed equally to a shallow benthic (sedimentary seafloor at 6 m water depth) and a pelagic habitat (3 m water depth) to compare the impact of these different environments on fouling and degradation. The amount of biofilm increased on both plastic types and in both habitats. The diatom abundance and diversity differed significantly between the habitats and the plastic types. Diatoms were more abundant on samples from the pelagic zone. We anticipate that specific surface properties of the polymer types induced different biofilm communities on both plastic types. Additionally, different environmental conditions between the benthic and pelagic experimental site such as light intensity and shear forces may have influenced unequal colonisation between these habitats. The oxygen production rate was negative for all samples, indicating that the initial biofilm on marine plastic litter consumes oxygen, regardless of the plastic type or if exposed in the pelagic or the benthic zone. Mechanical tests did not reveal degradation within one month of exposure. However, scanning electron microscopy (SEM) analysis displayed potential signs of degradation on the plastic surface, which differed between both plastic types. This study indicates that the early biofilm formation and composition are affected by the plastic type and habitat. Further, it reveals that already within two weeks biodegradable plastic shows signs of degradation in the benthic and pelagic habitat.     

地膜降解途径及机理研究进展

我国地膜使用量已占农业塑料薄膜使用量的半数以上,其大规模使用在带来巨大经济价值的同时也造成了“白色污染”,残膜难回收、难降解,直接影响土壤的再生能力。本文综述了我国地膜使用现状及其降解方面的研究进展,分别对地膜降解的生物与非生物途径和机理进行了概括,重点分析了非生物降解途径中光催化、金属离子掺杂等对聚烯烃降解的协同效应,以及生物降解途径中不同来源的菌和酶对聚烯烃降解效率的影响,并展望未来发展出更为高效的地膜降解方式,为后续地膜降解及环境中微塑料的降解研究提供参考。     

Kinetics and mechanism of heterogeneous hydrolysis of poly[(R)-3-hydroxybutyrate] film by PHA depolymerases

The kinetics and mechanism of enzymatic degradation on the surface of poly[(R)-3-hydroxybutyrate] (P[(R)-3HB]) film have been studied using three types of extracellular poly(hydroxyalkanoate) (PHA) depolymerases from Alcaligenes faecalis, Pseudomonas pickettii and Comamonas testosteroni. The monomer and dimer of 3-hydroxybutyric acid were produced during the course of the enzymatic degradation of P[(R)-3HB] film, and the rate of production was determined by monitoring the increase in absorbance at 210 nm on a spectrophotometer. The rate of enzymatic degradation increased to a maximum value with the concentration of PHA depolymerase, followed by a gradual decrease. The kinetic data were accounted for in terms of a heterogeneous enzymatic reaction, involving enzymatic degradation on the surface of P[(R)-3HB] film via two steps of adsorption and hydrolysis by a PHA depolymerase with binding and catalytic domains. The kinetic results suggest that the properties of the catalytic domains are very similar among the three PHA depolymerases, but that those of the binding domains are strongly dependent on the type of depolymerase.     

Economic comparison of enzymatic reactors and advanced oxidation processes applied to the degradation of phenol as a model compound

Abstract

Some micropollutants present in wastewaters are barely removed in sewage treatment plants. In many cases a post-treatment process based on separation and/or oxidation has to be applied. The aim of this study was the technical and economic comparison of enzymatic technologies with other advanced oxidation processes (AOPs) for the degradation of phenol. Batch and continuous enzymatic reactors, using free and immobilized manganese peroxidase (MnP, EC 1.11.1.13), were considered. Continuous degradation of phenol in an enzymatic membrane reactor was shown to be the fastest process and degradation in a continuous reactor with immobilized enzyme involved the lowest consumption of enzyme. However, the immobilization process increased the enzyme cost 100-fold. A continuous enzymatic membrane reactor gave high degradation efficiency and may be a viable technology for phenol removal when compared with other AOPs from both technical and economic points of view.     

Digestive capacities of leaf-cutting ants and the contribution of their fungal cultivar to the degradation of plant material

Leaf-cutting ants (tribe Attini) are a unique group of ants that cultivate a fungus that serves as a main source of their food. The fungus is grown on fresh leaves that are harvested by workers. We examine the respective contribution of ants and their symbiotic fungus in the degradation of plant material by examining the digestive capacities of seven Attini species in the genera Atta and Acromyrmex. The results show that both, the ants and their mutualistic fungi, have complementary enzymatic activities. Ants are specialized in the degradation of low molecular weight substrates (oligosaccharides and heterosides) whereas the fungus displays high polysaccharidase activity. The two genera Atta and Acromyrmex are not distinguished by a specific enzymatic activity. The seven different mutualistic associations examined display a similar enzymatic profile but have quantitative differences in substrate degradation activities. The respective contribution of ants and the fungus garden in plant degradation are discussed.     

Spectroscopic study on the enzymatic degradation of a biodegradable composite periodontal membrane

The enzymatic in vitro degradation of a commercial biodegradable hydroxyapatite (HA)-polymer (poly(epsilon-caprolactone)-poly(oxyethylene)(POE)-poly(epsilon-caprolactone) block copolymer) composite membrane was investigated by Raman and IR spectroscopies in two enzymatic solutions at 37 degrees C: esterase and alpha-chymotrypsin in saline phosphate buffer (SPB, pH 7.4). The degradation was found to be faster in the enzymatic medium than in SPB and alkaline solutions. The fastest degradation rate was observed in esterase solution. The trend of properly chosen Raman and IR intensity ratios was evaluated to go deeper inside the degradation mechanism: both polymeric and apatitic components were found to be involved in degradation. The former underwent preferential degradation of POE blocks, while HA is removed by the degradation medium faster than the polymer. Vibrational spectroscopy proved a valid tool for investigating the degradation of the membrane.     

评论：塑料结合模块促进聚对苯二甲酸乙二醇酯的酶法降解

塑料的大量生产和无节制的使用已造成严重的环境污染。为了减少塑料废物对环境的影响,近年来塑料酶法降解已成为国内外研究者关注的热点。例如,通过蛋白质工程策略提高塑料降解酶催化活性和热稳定性,进一步提高酶法降解的效率。另外,通过融合酶策略将塑料结合模块与塑料降解酶融合,也可以促进塑料降解。近期发表在期刊Chem Catalysis的一项研究表明,采用碳水化合物结合模块融合策略可以在低浓度(<10 wt%)的底物聚对苯二甲酸乙二醇酯[poly(ethylene terephthalate),PET]中提高塑料降解酶的活性。但是在高浓度底物(10 wt%−20 wt%)中,该策略无法提高PET的酶法降解。该项研究对于采用塑料结合模块促进酶法降解塑料具有重要的指导意义。     

Enzymatic degradation of low soluble compounds in monophasic water: solvent reactors. Kinetics and modeling of anthracene degradation by MnP

Polycyclic aromatic hydrocarbons (PAHs) are toxic compounds presenting low water solubility and high hydrophobicity, which greatly hampers their natural biodegradation. The enzymatic degradation of a model compound, anthracene, was evaluated in presence of a miscible solvent for an increased solubility. Manganese peroxidase, a ligninolytic enzyme from white-rot fungi, was used as biocatalyst in a medium containing acetone. The kinetic parameters of the enzymatic degradation of anthracene, obtained from fed-batch experiments, were applied to model the operation of a continuous reactor. Kinetics comprised a Michaelis-Menten equation, modified with an autocatalytic term, assumed to the effect of quinones acting as electron carriers, and a logistic function related to enzyme activity. The continuous reactor has been operated for 108 h, attaining a 90% of anthracene degradation, which demonstrated the feasibility of the system for its application in the removal of poorly soluble compounds. The model of this reactor permitted to predict accurately anthracene degradation in different conditions, such as external addition of anthraquinone and different enzymatic activities.     

Enzymatic degradation of hydroxypropyltrimethylammonium wheat starches

The enzymatic degradation of hydroxypropyltrimethylammonium modified starches synthesised by dry process was compared with that of hydroxypropyltrimethylammonium modified starches synthesised in glycerol–water plasticised molten medium. The enzymatic degradation rate of products from both origins decreased as the degree of substitution increased. However, two distinct enzymatic degradation profiles were obtained. Dry process products displayed a regular decrease pattern as DS increased. Molten medium synthesised cationic starches displayed a constant degradation level on a wide DS range with ,β-amylase and amyloglucosidase, whereas isoamylase degradation rapidly reached its degradation limit at DSs 0.05. The various plasticising conditions used to synthesise cationic starch in molten medium show no influence on the enzymatic degradation.

By measuring the affinity of -amylase, β-amylase and isoamylase for native, extruded non-modified and hydroxypropyltrimethylammonium-modified starches. It was evident that the enzymes’ affinity for the substrate diminishes with increasing chemical modification, particularly in the case of -amylase, suggesting that the location of cationic groups impairs the enzyme’s recognition of the substrate. Structural elements of limit dextrins were analysed by ¹H NMR.     

Reactor design for minimizing product inhibition during enzymatic lignocellulose hydrolysis: I. Significance and mechanism of cellobiose and glucose inhibition on cellulolytic enzymes

Achievement of efficient enzymatic degradation of cellulose to glucose is one of the main prerequisites and one of the main challenges in the biological conversion of lignocellulosic biomass to liquid fuels and other valuable products. The specific inhibitory interferences by cellobiose and glucose on enzyme-catalyzed cellulose hydrolysis reactions impose significant limitations on the efficiency of lignocellulose conversion — especially at high-biomass dry matter conditions. To provide the base for selecting the optimal reactor conditions, this paper reviews the reaction kinetics, mechanisms, and significance of this product inhibition, notably the cellobiose and glucose inhibition, on enzymatic cellulose hydrolysis. Particular emphasis is put on the distinct complexity of cellulose as a substrate, the multi-enzymatic nature of the cellulolytic degradation, and the particular features of cellulase inhibition mechanisms and kinetics. The data show that new strategies that place the bioreactor design at the center stage are required to alleviate the product inhibition and in turn to enhance the efficiency of enzymatic cellulose hydrolysis. Accomplishment of the enzymatic hydrolysis at medium substrate concentration in separate hydrolysis reactors that allow continuous glucose removal is proposed to be the way forward for obtaining feasible enzymatic degradation in lignocellulose processing.     

Hypoxia-inducible factor 1α under rapid enzymatic hypoxia: Cells sense decrements of oxygen but not hypoxia per se

HIF1 (hypoxia-inducible factor 1α) is considered a central oxygen-threshold sensor in mammalian cells. In the presence of oxygen, HIF1 is marked by prolyl hydroxylases (PHDs) at the oxygen-dependent degradation (ODD) domain for ubiquitination followed by rapid proteasomal degradation. However, the actual mechanisms of oxygen sensing by HIF1 are still controversial. Thus, HIF1 expression correlates poorly with tissue oxygen levels, and PHDs are themselves target genes of HIF1 considered to readjust to new oxygen thresholds. In contrast to hypoxia chambers, we here establish an enzymatic model that allows both the rapid induction of stable hypoxia and independent control of H₂O₂. Rapid enzymatic hypoxia only transiently induced HIF1 in various cell types and the HIF1 was completely degraded within 8–12 h despite sustained hypoxia. HIF1 degradation under sustained hypoxia could be blocked by a competitive ODD–GFP construct and PHD siRNA, but also by cobalt chloride and micromolar H₂O₂ levels. Concomitant induction of PHDs further confirmed their role in degrading HIF1 under enzymatic hypoxia. The rapid and complete degradation of HIF1 under enzymatic hypoxia suggests that, in addition to hypoxia sensing, the HIF1/PHD loop may rather compensate for fluctuations of tissue oxygen staying tuned to other, e.g., metabolic, signals. In addition to hypoxia chambers, enzymatic hypoxia provides a valuable tool for independently studying the regulatory functions of hypoxia and oxidative stress in vitro.     

微生物降解塑料的研究进展

塑料广泛应用于人类的生活中,其中约80%的塑料垃圾被填埋,最终成为陆地和海洋垃圾。由于管理与处置不善,这些废弃物造成了巨大的环境污染,目前回收再利用是较好的处置方式,但对某些塑料废弃物并没有妥善的处置方式。生物降解作为环境友好的处置方式,具有巨大的应用潜力。本文对聚对苯二甲酸乙二醇酯、聚乙烯、聚氯乙烯、聚丙烯、聚苯乙烯和聚氨酯这6种常用塑料的降解微生物及生物降解机制进行了总结,对目前微生物降解塑料存在的问题进行了分析,并提出了促进微生物降解塑料应用的途径,为生物降解塑料菌株和降解酶的开发应用、降解机制研究提供理论参考。     

Biodegradation of a polyvinyl alcohol-starch blend plastic film

Attempts were made to elucidate the degradation mechanism of a polyvinyl alcohol (PVA)-starch blend plastic. A part of the starch fraction of this plastic was dissolved into an aqueous phase in a control test. Treatment with a PVA-degrading bacterium or enzyme gave a maximal weight loss of approximately 70% and film breakage occurred. Since this plastic contains 40% PVA, it is apparent that not only the PVA fraction but also a considerable portion of the starch fraction was lost from the film by treatment with the PVA-degrading enzyme. As the PVA-degrading bacterium and enzyme used here showed no starch-degrading activity, loss of the starch fraction seems to depend on its dissolution with degradation of the PVA fraction. These experimental results indicated that the degradation of the PVA fraction is an important requisite for complete degradation or decomposition of this plastic film.     

Degradation of Oxo-Biodegradable Plastic by Pleurotus ostreatus

Growing concerns regarding the impact of the accumulation of plastic waste over several decades on the environmental have led to the development of biodegradable plastic. These plastics can be degraded by microorganisms and absorbed by the environment and are therefore gaining public support as a possible alternative to petroleum-derived plastics. Among the developed biodegradable plastics, oxo-biodegradable polymers have been used to produce plastic bags. Exposure of this waste plastic to ultraviolet light (UV) or heat can lead to breakage of the polymer chains in the plastic, and the resulting compounds are easily degraded by microorganisms. However, few studies have characterized the microbial degradation of oxo-biodegradable plastics. In this study, we tested the capability of Pleurotus ostreatus to degrade oxo-biodegradable (D₂W) plastic without prior physical treatment, such as exposure to UV or thermal heating. After 45 d of incubation in substrate-containing plastic bags, the oxo-biodegradable plastic, which is commonly used in supermarkets, developed cracks and small holes in the plastic surface as a result of the formation of hydroxyl groups and carbon-oxygen bonds. These alterations may be due to laccase activity. Furthermore, we observed the degradation of the dye found in these bags as well as mushroom formation. Thus, P. ostreatus degrades oxo-biodegradable plastics and produces mushrooms using this plastic as substrate.