Ligninases

  • 文章类型: Journal Article
    堆肥涉及选择能够抵抗过程中产生的高温并降解木质纤维素的微生物群。深入了解参与这种生物转化的嗜热微生物群落对于提高堆肥效率和为生物精炼提供热稳定的生物质降解酶是有价值的。这项研究调查了在植物废物堆肥的所有阶段降解木质纤维素的嗜热微生物培养,专注于动态,酶,以及这样一个社区的每个成员的耐热性。结果表明,堆肥结束时,58%的全纤维素(纤维素加半纤维素)和7%的木质素被降解。整个真菌嗜热种群表现出降解木质纤维素的活性,而大约8-10%的嗜热细菌具有这种特征,虽然专门用于半纤维素降解(木聚糖降解)。由于两组的患病率,它们的酶活性,和广泛的耐热性,在整个过程中,它们在半纤维素的分解中起着关键作用,而纤维素和木质素的降解仅限于在过程结束时持续存在的一些嗜热真菌的活性。木聚糖分解细菌分离株(159株)主要包括Firmicutes成员(96%)以及放线菌(2%)和变形菌(2%)的一些代表。最普遍的物种是地衣芽孢杆菌和苍白气杆菌。嗜热真菌(27株)仅包括4种,也就是蓝藻热霉菌,嗜热滑石,烟曲霉,和Gibellulopsisnigrescens,其中烟曲霉和羊毛虫占主导地位。同一物种的几个菌株在堆肥阶段明显进化,表现出具有不同耐热性和新酶表达的表型,即使以前没有描述过这个物种,作为对不断变化的堆肥环境的回应。热淀粉芽孢杆菌菌株,热降氮地芽孢杆菌,T.lanuginosus,和表现出相当酶活性的烟曲霉被选作产生热酶的潜在候选物。本研究为进一步研究高温木质纤维素分解微生物在堆肥过程中适应和获得新性状的机制及其在生物技术加工中的潜在用途奠定了基础。
    Composting involves the selection of a microbiota capable of resisting the high temperatures generated during the process and degrading the lignocellulose. A deep understanding of the thermophilic microbial community involved in such biotransformation is valuable to improve composting efficiency and to provide thermostable biomass-degrading enzymes for biorefinery. This study investigated the lignocellulose-degrading thermophilic microbial culturome at all the stages of plant waste composting, focusing on the dynamics, enzymes, and thermotolerance of each member of such a community. The results revealed that 58% of holocellulose (cellulose plus hemicellulose) and 7% of lignin were degraded at the end of composting. The whole fungal thermophilic population exhibited lignocellulose-degrading activity, whereas roughly 8-10% of thermophilic bacteria had this trait, although exclusively for hemicellulose degradation (xylan-degrading). Because of the prevalence of both groups, their enzymatic activity, and the wide spectrum of thermotolerance, they play a key role in the breakdown of hemicellulose during the entire process, whereas the degradation of cellulose and lignin is restricted to the activity of a few thermophilic fungi that persists at the end of the process. The xylanolytic bacterial isolates (159 strains) included mostly members of Firmicutes (96%) as well as a few representatives of Actinobacteria (2%) and Proteobacteria (2%). The most prevalent species were Bacillus licheniformis and Aeribacillus pallidus. Thermophilic fungi (27 strains) comprised only four species, namely Thermomyces lanuginosus, Talaromyces thermophilus, Aspergillus fumigatus, and Gibellulopsis nigrescens, of whom A. fumigatus and T. lanuginosus dominated. Several strains of the same species evolved distinctly at the stages of composting showing phenotypes with different thermotolerance and new enzyme expression, even not previously described for the species, as a response to the changing composting environment. Strains of Bacillus thermoamylovorans, Geobacillus thermodenitrificans, T. lanuginosus, and A. fumigatus exhibiting considerable enzyme activities were selected as potential candidates for the production of thermozymes. This study lays a foundation to further investigate the mechanisms of adaptation and acquisition of new traits among thermophilic lignocellulolytic microorganisms during composting as well as their potential utility in biotechnological processing.
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  • 文章类型: Journal Article
    细菌系统在木质纤维素生物质的解聚中得到了广泛的关注,由于其高度的功能多样性和适应性。为了在生物精炼厂内实现木质纤维素残留物的充分微生物利用和生物产品的成本效益生产,需要多种代谢途径和各种特异性的酶。在这项工作中,高度多样化的有氧,从Keri湖富集的中温细菌,生物质降解增加和天然地下石油泄漏的原始沼泽,探索了它们对木质纤维素底物的代谢多功能性和酶促潜力。大量的假单胞菌,从有机溶剂木质素作为唯一的碳和能源的富集培养中获得,能够吸收一系列木质素相关的芳香族化合物。比较复杂的细菌聚生体,包括放线菌的成员,变形杆菌,芽孢杆菌,鞘杆菌属,还有黄杆菌,还从具有木聚糖或羧甲基纤维素作为唯一碳源的培养物中富集。通过表达对结晶或无定形纤维素和木聚糖的水解活性,许多单独的分离株可以靶向不同结构的木质纤维素多糖。特定的分离株显示出由碱预处理的农业废物制备的木质素水解产物的生长潜力增加。结果表明,Keri分离株代表了有效的木质纤维素降解物库,具有在木质纤维素生物炼制中工业应用的巨大潜力。
    Bacterial systems have gained wide attention for depolymerization of lignocellulosic biomass, due to their high functional diversity and adaptability. To achieve the full microbial exploitation of lignocellulosic residues and the cost-effective production of bioproducts within a biorefinery, multiple metabolic pathways and enzymes of various specificities are required. In this work, highly diverse aerobic, mesophilic bacteria enriched from Keri Lake, a pristine marsh of increased biomass degradation and natural underground oil leaks, were explored for their metabolic versatility and enzymatic potential towards lignocellulosic substrates. A high number of Pseudomonas species, obtained from enrichment cultures where organosolv lignin served as the sole carbon and energy source, were able to assimilate a range of lignin-associated aromatic compounds. Comparatively more complex bacterial consortia, including members of Actinobacteria, Proteobacteria, Bacilli, Sphingobacteria, and Flavobacteria, were also enriched from cultures with xylan or carboxymethyl cellulose as sole carbon sources. Numerous individual isolates could target diverse structural lignocellulose polysaccharides by expressing hydrolytic activities on crystalline or amorphous cellulose and xylan. Specific isolates showed increased potential for growth in lignin hydrolysates prepared from alkali pretreated agricultural wastes. The results suggest that Keri isolates represent a pool of effective lignocellulose degraders with significant potential for industrial applications in a lignocellulose biorefinery.
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  • 文章类型: Journal Article
    BACKGROUND: Textile industry represents one prevalent activity worldwide, generating large amounts of highly contaminated and rich in azo dyes wastewater, with severe effects on natural ecosystems and public health. However, an effective and environmentally friendly treatment method has not yet been implemented, while concurrently, the increasing demand of modern societies for adequate and sustainable energy supply still remains a global challenge. Under this scope, the purpose of the present study was to isolate promising species of yeasts inhabiting wood-feeding termite guts, for combined azo dyes and textile wastewater bioremediation, along with biodiesel production.
    RESULTS: Thirty-eight yeast strains were isolated, molecularly identified and subsequently tested for desired enzymatic activity, lipid accumulation, and tolerance to lignin-derived metabolites. The most promising species were then used for construction of a novel yeast consortium, which was further evaluated for azo dyes degradation, under various culture conditions, dye levels, as well as upon the addition of heavy metals, different carbon and nitrogen sources, and lastly agro-waste as an inexpensive and environmentally friendly substrate alternative. The novel yeast consortium, NYC-1, which was constructed included the manganese-dependent peroxidase producing oleaginous strains Meyerozyma caribbica, Meyerozyma guilliermondii, Debaryomyces hansenii, and Vanrija humicola, and showed efficient azo dyes decolorization, which was further enhanced depending on the incubation conditions. Furthermore, enzymatic activity, fatty acid profile and biodiesel properties were thoroughly investigated. Lastly, a dye degradation pathway coupled to biodiesel production was proposed, including the formation of phenol-based products, instead of toxic aromatic amines.
    CONCLUSIONS: In total, this study might be the first to explore the application of MnP and lipid-accumulating yeasts for coupling dye degradation and biodiesel production.
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  • 文章类型: Journal Article
    如今,回收生物质是必不可少的,这不仅是因为化石能源逐渐枯竭,还因为环境污染,由于能源使用的增加,必须减少。本文拟对目前使用的植物生物质处理方法的结果进行综述。我们的目的还在于审查目前尚未使用的已发布方法。旨在探索用于生物质回收的新方法和酶的可能性。本概述的结果几乎在每个领域都令人困惑。在生物质的预处理以及所使用的酶的多样性和应用方面已经取得了进展。基于分子建模,在生物质加工过程中,在改变现有酶的功能和适应环境条件方面进展甚微。几乎没有任何出版物使用分子建模技术来改善酶功能并使酶适应各种环境条件。我们的观点是使用现代计算,生物化学,和生物技术方法将能够有目的地设计更有效和适用于生物质加工的酶。
    Recycling biomass is indispensable these days not only because fossil energy sources are gradually depleted, but also because pollution of the environment, caused by the increasing use of energy, must be reduced. This article intends to overview the results of plant biomass processing methods that are currently in use. Our aim was also to review published methods that are not currently in use. It is intended to explore the possibilities of new methods and enzymes to be used in biomass recycling. The results of this overview are perplexing in almost every area. Advances have been made in the pre-treatment of biomass and in the diversity and applications of the enzymes utilized. Based on molecular modeling, very little progress has been made in the modification of existing enzymes for altered function and adaptation for the environmental conditions during the processing of biomass. There are hardly any publications in which molecular modeling techniques are used to improve enzyme function and to adapt enzymes to various environmental conditions. Our view is that using modern computational, biochemical, and biotechnological methods would enable the purposeful design of enzymes that are more efficient and suitable for biomass processing.
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  • 文章类型: Journal Article
    Liometopum apiculatum is a species of ants widely distributed in arid and semi-arid ecosystems where there is a relative food shortage compared with tropical ecosystems. L. apiculatum has established an ecological balance involving symbiotic interactions, which have allowed them to survive through mechanisms that are still unknown. Therefore, the aim of this study was to explore the metabolic potential of isolated bacteria from L. apiculatum using enzymatic activity assay and substrate assimilation. Results revealed a complex bacteria consortium belonging to Proteobacteria, Firmicutes, and Actinobacteria phylum. Most of the isolated bacteria showed activities associated with biopolymers degradation, from them Exiguobacterium and B. simplex showed the highest amylolytic activity (27 U/mg protein), while A. johnsonii and B. pumulis showed the highest cellulolytic and xylanolytic activities (1 and 2.9 U/mg protein, respectively). By other hand, some microorganisms such as S. ficaria, E. asburiae, P. agglomerans, A. johnsonii, S. rubidaea, S. marcescens, S. warneri, and M. hydrocarbonoxydans were able to grow up to 1000 mg/L of phthalates esters. These results not only revealed the important contribution of the symbionts in L apiculatum ants feeding habits, but also have shown a promising source of enzymes with potential biotechnological applications such as lignocellulosic biomass hydrolysis and bioremediation processes.
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  • 文章类型: Journal Article
    微生物活性是红树林养分循环的主要途径。总的来说,微生物具有降解木质纤维素化合物的能力。在来自红树林的微生物群的生物技术潜力中,值得注意的是内生真菌,可以被认为是不同生物活性化合物的有效来源。在这个意义上,从红树林采样Cananeia中分离出三十(30)种内生真菌,SP,巴西。分析了这些微生物的酶活性,包括:木质素过氧化物酶EC1.11.1.14,锰过氧化物酶EC1.11.1.13和漆酶EC1.10.3.2,以及内切纤维素酶EC3.2.1.4和内切木聚糖酶EC3.2.1.8。除此之外,还研究了生物活性次级代谢产物如生物表面活性剂和/或生物乳化剂的产生。作为结果,选择了十九(19)个分离株的木质素分解能力,其中九(9)有关纤维素酶活性和十三(13)显示木聚糖酶能力。真菌分离株命名为3(3),特征为镰刀菌,显示出突出的木质素过氧化物酶(42.4UL-1)和锰过氧化物酶(23.6UL-1)活性。分离株63.1,也与镰刀菌有关。属,选择其漆酶活性(41.5UL-1)。从所有被调查的真菌中,选择分离物47(4)木霉的纤维素分解和木聚糖分解活性,分别显示45.23和26.09UmL-1。相同的真菌还显示出生物表面活性剂能力,其表面张力降低至38mN/m。此外,十五(15)种真菌表现出生物乳化剂活性,E24值高达62.8%。
    Microbial activity is the main route for cycling mangrove nutrients. In general, microorganisms have abilities to degrade lignocellulosic compounds. Among the biotechnological potential of the microbiota from mangroves, it is noteworthy about endophytic fungi, which can be considered as effective sources of different bioactive compounds. In this sense, thirty (30) endophytic fungi were isolated from mangrove forest sampling Cananeia, SP, Brazil. These microorganisms were analyzed about their enzymatic activities including: lignin peroxidase EC 1.11.1.14, manganese peroxidase EC 1.11.1.13 and laccase EC 1.10.3.2, as well endo-cellulase EC 3.2.1.4 and endo-xylanase EC 3.2.1.8. Besides that, production of bioactive secondary metabolites like biosurfactant and/or bioemulsifier was also investigated. As results, nineteen (19) isolates were selected about their ligninolytic abilities, nine (9) of them about cellulase activity and thirteen (13) showed xylanase abilities. The fungal isolate named as 3(3), characterized as Fusarium sambucinum, showed a prominent lignin peroxidase (42.4 U L-1) and manganese peroxidase (23.6 U L-1) activities. The isolate 63.1, also related to Fusarium sp. genera, was selected about its laccase activity (41.5 U L-1). From all the investigated fungi, the isolate 47(4) Trichoderma camerunense was selected about its cellulolytic and xylanolytic activities, showing 45.23 and 26.09 U mL-1, respectively. The same fungi also showed biosurfactant ability demonstrated by superficial tension decreasing to 38 mN/m. In addition, fifteen (15) fungi exhibited bioemulsifier activity, with E24 values up to 62.8%.
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  • 文章类型: Journal Article
    The ligninolytic enzyme consortium is one of the most-efficient oxidative systems found in nature, playing a pivotal role during wood decay and coal formation. Typically formed by high redox-potential oxidoreductases, this array of enzymes can be used within the emerging lignocellulose biorefineries in processes that range from the production of bioenergy to that of biomaterials. To ensure that these versatile enzymes meet industry standards and needs, they have been subjected to directed evolution and hybrid approaches that surpass the limits imposed by nature. This Opinion article analyzes recent achievements in this field, including the incipient groundbreaking research into the evolution of resurrected enzymes, and the engineering of ligninolytic secretomes to create consolidated bioprocessing microbes with synthetic biology applications.
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  • 文章类型: Journal Article
    Fungi have been recently recognized as organisms able to grow in presence of high salt concentration with halophilic and halotolerance properties and their ligninolytic enzyme complex have an unspecific action enabling their use to degradation of a number of xenobiotic compounds. In this work, both the effect of salt and polyols on growth of the basidiomycetes strains, on their ability to produce ligninolytic enzyme and diuron degradation were evaluated. Results showed that the presence of NaCl in the culture medium affected fungal specimens in different ways. Seven out of ten tested strains had growth inhibited by salt while Dacryopinax elegans SXS323, Polyporus sp MCA128 and Datronia stereoides MCA167 fungi exhibited higher biomass production in medium containing 0.5 and 0.6 mol.L(-1) of NaCl, suggesting to be halotolerant. Polyols such as glycerol and mannitol added into the culture media improved the biomass and ligninases production by D. elegans but the fungus did not reveal consumption of these polyols from media. This fungus degraded diuron in medium control, in presence of NaCl as well as polyols, produced MnP, LiP and laccase.
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