聚对苯二甲酸乙二醇酯(PET)降解酶的研究进展

聚对苯二甲酸乙二醇酯(Polyethylene terephthalate,PET)因其优越的物理化学性质,在各个领域尤其在包装产业得到了广泛的应用.然而,由于使用后的PET处置不当,对生态环境造成了严重威胁.目前生物降解尤其是酶促降解已成为极具可行性且环境友好的PET处理方式.本文集中梳理和总结了近年来已报道的PET...     

聚对苯二甲酸乙二醇酯降解酶的研究进展

聚对苯二甲酸乙二醇酯(Polyethylene terephthalate,PET)因其良好的耐用性和可塑性,已在世界范围内的工业领域和日常生活中得到广泛应用。目前自然环境中大量PET使用废弃物的积累和迁移给全球生态系统带来了严重负担,因此PET的降解问题已成为全球性的热点问题。微生物酶降解法目前被认为是一种理想绿色PET降解方法,有希望应用于大规模降解PET废弃物降解处理。传统的PET降解酶主要包括脂肪酶、酯酶和角质酶等,但这些酶的PET降解活性相对不高。近期科学家从Ideonella sakaiensis细菌中分离了一种新型水解酶PETase,能够特异性高效降解PET。本文从结构生物学角度对多种PET降解酶进行梳理,重点总结了新近发现的PETase催化机制,为发展改造更有效的PET降解酶提供理论依据。     

碳水化合物结合模块-嗜热子囊菌角质酶融合蛋白在PET降解中的应用

随着全球经济的发展,聚对苯二甲酸乙二醇酯(PET)塑料的生产量大幅提高,其废弃总量也逐年递增.废弃PET处理方法主要包括填埋、焚烧及生物降解等.填埋和焚烧均会造成二次污染,而生物降解因其环境友好特性逐渐成为研究热点.相关研究表明,碳水化合物结合模块(carbohydrate binding module,CBM)可以有...     

聚对苯二甲酸乙二醇酯水解酶IsPETase及其应用展望

随着生物技术的迅速发展,酶解法作为一种绿色可持续的聚对苯二甲酸乙二醇酯(polyethylene terephthalate, PET)回收处理方案,有望解决全球范围内废弃PET带来的环境污染问题。众多PET水解酶中,来自Ideonella sakaiensis的PETase因其对PET底物的高特异性成为当下研究的热点。基于对酶的结构和功能的深刻理解,本文总结了近年来PETase的工程改造进展,以提高酶的降解活性、热稳定性和对底物的吸附性;介绍了PETase的分泌表达策略、细胞表面展示技术,以及PETase与MHETase双酶系统的应用;最后,我们对塑料生物降解领域存在的挑战及可能的解决途径进行了展望,这些工作将为促进聚合物生物降解的实际应用提供参考。     

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。     

聚丁二酸丁二醇酯(PBS)的生物降解的研究进展

聚丁二酸丁二醇酯(poly(butylene succinate), PBS)是一种人工合成的脂肪族聚酯化合物。PBS的生产成本低、热稳定性好,具有良好加工性能、机械性能以及力学性能等优点。本文就近年来PBS在生物降解方面的研究进展进行了综述,具体包括PBS的生物堆肥降解、PBS的微生物降解以及PBS降解酶的相关研究。最后对PBS生物降解研究进展做出了总结。     

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。     

来源于海洋宏基因组塑料降解酶Ple629的耐热性提升改造

石化来源的聚酯类塑料如聚对苯二甲酸乙二醇酯(polyethylene terephthalate,PET)以及聚己二酸/对苯二甲酸丁二醇酯(polybutylene adipate terephthalate,PBAT)等已被广泛使用,但由于它们在自然界中难以降解或生物降解周期较长导致了严重的环境污染,因此对这些塑料废弃物的处理是亟待解决的问题之一。从循环经济的角度考虑,利用生物酶法对聚酯类塑料如PET或PBAT等的废弃物进行解聚,再将解聚产物进行循环利用,是一个很有潜力的研究方向。探究近年来关于聚酯塑料降解酶的报道发现,高活性且耐高温的降解酶会有更大的潜在优势。来自海洋微生物宏基因组的中温塑料降解酶Ple629,在常温下对聚酯类塑料PET和PBAT均有较好的降解活力,但由于不耐受高温,限制了其潜在应用。在前期获得Ple629三维结构的基础上,本研究基于结构比对及能量设计,找到了一些潜在提升其热稳定性的位点进行改造设计,并对突变体进行了表达纯化和热稳定性测定。突变体V80C和D226C/S281C的熔点温度(Tm)值分别提升了5.2℃和6.9℃,突变体D226C/S281C的活性也比野生型酶提高了1.5倍,为后续对Ple629的进一步改造提供了思路和依据。     

聚对苯二甲酸乙二醇酯水解酶研究进展

塑料自20世纪首次合成以来给人类生活带来了极大的便利。然而,塑料稳定的高分子结构导致了塑料废弃物的持续堆积,对生态环境和人类健康均造成严重威胁。聚对苯二甲酸乙二醇酯[poly(ethylene terephthalate),PET]是产量最高的一种聚酯类塑料,近年来PET水解酶的相关研究展现出生物酶法对塑料进行降解、回收的巨大潜力,也为塑料生物降解机制研究建立了参考范例。本文综述了不同微生物来源的PET水解酶及其PET降解能力,阐述了最具代表性的PET水解酶—IsPETase降解PET的催化机理,并总结了近年来通过酶工程改造而获得的高效降解酶,为未来的PET降解机制研究、PET高效降解酶的进一步挖掘和改造提供参考。     

一株嗜热聚对苯二甲酸乙二醇酯降解菌的分离及其降解特性解析北大核心

【目的】大量聚对苯二甲酸乙二醇酯(polyethylene terephthalate,PET)塑料作为废弃物被丢弃,严重危害生态健康。针对嗜热PET降解菌缺乏这一情况,本研究旨在获得能够降解PET的嗜热菌,并阐述其降解机制。【方法】采集云南腾冲热泉中的废弃PET瓶,分析其表面生物膜的微生物群落多样性,从中筛选能够以PET为营养源生长的嗜热菌,并基于16S rRNA基因序列加以鉴定;以菌株的定殖能力与生长曲线为指标,优选出降解能力较强的降解菌,并测定其最适pH、温度和NaCl浓度;降解能力较强的降解菌分别作用于PET及PET中间体双(羟乙基)对苯二甲酸酯[bis(hydroxyethyl)terephthalate,BHET]和对苯二甲酸单(2-羟乙基)酯[mono(2-hydroxyethyl)terephthalate,MHET],测定产物生成量与降解率;通过观察PET膜表面微观结构、活菌数、酯酶活性等探究降解菌与PET的互作过程。【结果】废弃PET瓶表面生物膜中的微生物群落多样性低;从生物膜中筛选出5株能够以PET为营养源生长的嗜热菌;其中,菌株JQ3以PET为唯一碳源生长最佳,作为降解能力较强的降解菌,被鉴定为嗜热淀粉芽孢杆菌(Bacillus thermoamylovorans),其最适生长pH为7.0、最适生长温度为50℃、最适生长NaCl浓度为0.5%;菌株JQ3以0.043 mg PET/d的速率降解PET,对苯二甲酸(terephthalic acid,TPA)产量在第7天达到峰值45.2 mmol/L;菌株JQ3对PET中间体降解效率显著,6 h可降解85.9%的BHET,60 h可降解50.1%的MHET。菌株JQ3能够定殖于PET表面并形成生物膜,侵蚀PET并造成开裂和剥落。【结论】B.thermoamylovorans JQ3作为一株嗜热PET降解菌,能够高温(60℃)降解PET及其中间体,为实现PET的有效降解提供了新策略。     

重组角质酶的发酵制备及其对涤纶纤维的表面改性

对大肠杆菌表达嗜热子囊菌Thermobifida fusca角质酶的摇瓶诱导条件及3 L发酵罐扩大培养进行了研究,并探讨了角质酶对涤纶纤维的改性作用。结果表明,在摇瓶培养中,采用工业级TB培养基,用2 g/L乳糖诱导,菌体培养至对数生长前期添加0.5％甘氨酸,角质酶产量可达到128 U/mL。在3 L发酵罐扩大培养中,补料培养生物量 (OD600) 最大达到35,角质酶酶活最高达506 U/mL,是迄今国内外报道细菌来源角质酶的最高水平。紫外分光光度法分析初步表明涤纶纤维经角质酶水解产生了对苯二甲酸类物质     

Facile preparation of biodegradable chitosan derivative having poly(butylene glycol adipate) side chains

Various modes are being explored for the construction of functional materials from nanoparticles. Despite these efforts, the assembly of nanoparticles remains challenging with respect to the requirement of multiple component organization on varying dimensions and length scales. The graft copolymers of chitosan with poly(butylene glycol adipate) (PBGA) were prepared due to the esterification reaction between PBGA and 6-O-succinate-N-phthaloyl-chitosan (PHCSSA) in the presence of toluene as a swelling agent. The graft copolymers are nanoparticles with the size of few hundred nanometers as observed from TEM. It is a potential method to combine chitosan with the hydrophobic synthetic polymers. The grafting reactions were conducted with various PBGA/PHCSSA feed ratios to obtain chitosan-g-PBGA copolymers with various PBGA contents. FT-IR, NMR, XRD, spectrofluorophotometer, and TEM were detected to characterize the copolymers.     

Degradation of microbial polyester poly(3-hydroxybutyrate) in environmental samples and in culture

Poly(3-hydroxybutyrate) [P(3HB)] test-pieces prepared from the polymer produced by Azotobacter chroococcum were degraded in natural environments like soil, water, compost and sewage sludge incubated under laboratory conditions. Degradation in terms of % weight loss of the polymer was maximum (45%) in sewage sludge after 200 days of incubation at 30°C. The P(3HB)-degrading bacterial cultures (36) isolated from degraded test-pieces showed different degrees of degradation in polymer overlayer method. The extent of P(3HB) degradation increases up to 12 days of incubation and was maximum at 30°C for majority of the cultures. For most efficient cultures the optimum concentration of P(3HB) for degradation was 0.3% (w/v). Supplementation of soluble carbon sources like glucose, fructose and arabinose reduced the degradation while it was almost unaffected with lactose. Though the cultures degraded P(3HB) significantly, they were comparatively less efficient in utilizing copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate [P(3HB-co-3HV)].     

Study of biodegradable polylactide/poly(butylene adipate-co-terephthalate) blends

Both polylactide (PLA) and poly(butylene adipate-co-terephthalate) (PBAT) are biodegradable polymers. They are thermoplastics which can be processed using most conventional polymer processing methods. PLA is high in strength and modulus (63 MPa and 3.4 GPa, respectively) but brittle (strain at break 3.8%) while PBAT is flexible and tough (strain at break approximately 710%). In view of their complementary properties, blending PLA with PBAT becomes a natural choice to improve PLA properties without compromising its biodegradability. In this study, PLA and PBAT were melt blended using a twin screw extruder. Melt elasticity and viscosity of the blends increased with the concentration of PBAT. Crystallization of the PLA component, phase morphology of the blend, mechanical properties, and toughening mechanism were investigated. The blend comprised an immiscible, two-phase system with the PBAT evenly dispersed in the form of approximately 300 nm domains within the PLA matrix. The PBAT component accelerated the crystallization rate of PLA but had little effect on its final degree of crystallinity. With the increase in PBAT content (5-20 wt %), the blend showed decreased tensile strength and modulus; however, elongation and toughness were dramatically increased. With the addition of PBAT, the failure mode changed from brittle fracture of the neat PLA to ductile fracture of the blend as demonstrated by tensile test and scanning electron microcopy (SEM) micrographs. Debonding between the PLA and PBAT domains induced large plastic deformation in PLA matrix ligaments.     

Biodegradation of poly(l-lactide)

The biodegradation of poly(L-lactide) (PLA) is reviewed. The important role of actinomycetes in PLA degradation is emphasized. These PLA-degrading actinomycetes belong phylogenetically to the Pseudonocardiaceae family and related genera, including Amycolatopsis, Lentzea, Streptoalloteichus, Kibdelosporangium and Saccharothrix. A PLA-degrading enzyme purified from an isolated Amycolatopsis strain-41 has substrate specificity on PLA higher than proteinase K. The application of these strains and their enzymes can be effectively used for biological treatment of plastic wastes containing PLA.     

Degradation of aliphatic polyester films by commercially available lipases with special reference to rapid and complete degradation of poly(L-lactide) film by lipase PL derived from Alcaligenes sp

Commercial lipases were examined for their degradation efficiency of aliphatic polyester films. In 100 days immersion of polyester films in lipase solutions at37 °C at pH 7.0,Lipase Asahi derived from Chromobacterium viscosum degraded polybutylene succinate-co-adipate (PBSA), poly (-caprolactone) (PCL) and polybutylene succinate (PBS), and Lipase F derived from Rhizopus niveus degraded PBSA and PCL during 4–17 days. Lipase F-AP15 derived fromRhizopus orizae could degrade PBSA in 22 days. In these cases, PBS and PBSA were mainly degraded to dimers, whereas PCL was mainly degraded to monomers. Only poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(PHB/V) and poly (L-lactide) (PLA) were not degraded in the experiments. However, PLA degraded completely at 55 °C, pH 8.5 with Lipase PL during 20 days. This result could be explained with the sequential reactions of the chemical hydrolysis of the polymer to oligomers at higher pH and temperature, and the succeeding enzymatic hydrolysis of oligomers to the monomers.     

聚3-羟基丁酸酯（PHB）生物降解过程的研究

利用DS9701菌株对聚3-羟基丁酸酯(PHB)膜进行降解,对降解到不同程度的PHB膜采用扫描电子显微镜观察其表面形态结构的变化,并对其降解产物进行分析测定.结果表明,PHB的生物降解首先发生在PHB表面的非晶部分,随后结晶部分开始降解,并且降解首先发生在球晶的中心部分.DS9701菌株所产生的PHB解聚酶主要降解PHB的第二个酯键,降解产物为二聚体.     

Enzymatic surface hydrolysis of poly(ethylene terephthalate) and bis(benzoyloxyethyl) terephthalate by lipase and cutinase in the presence of surface active molecules

A lipase from Thermomyces lanuginosus and cutinases from Thermobifida fusca and Fusarium solani hydrolysed poly(ethylene terephthalate) (PET) fabrics and films and bis(benzoyloxyethyl) terephthalate (3PET) endo-wise as shown by MALDI-Tof-MS, LC–UVD/MS, cationic dyeing and XPS analysis. Due to interfacial activation of the lipase in the presence of Triton X-100, a seven-fold increase of hydrolysis products released from 3PET was measured. In the presence of the plasticizer N,N-diethyl-2-phenylacetamide (DEPA), increased hydrolysis rates of semi-crystalline PET films and fabrics were measured both for lipase and cutinase. The formation of novel polar groups resulted in enhanced dye ability with additional increase in colour depth by 130% and 300% for cutinase and lipase, respectively, in the presence of plasticizer.