微生物降解秸秆木质素的研究进展

我国秸秆资源丰富，每年产生逾8亿t作物秸秆。通过秸秆直接还田或肥料化还田不仅可以减少化肥的施用量，缓解农业污染压力，还能实现农作物秸秆的循环利用。木质素结构复杂，且与纤维素和半纤维素相互缠绕，因此秸秆的自然腐解过程中，木质素是主要的限速因子，为了提高降解效率，木质素降解菌的发掘和降解机制也逐渐成为研究热点。本文综述了降解木质素的真菌和细菌的研究现状，对比其真菌和细菌降解特性的优缺点并分析复合降解菌群的优势。随后对木质素降解酶系的酶学性质、在不同微生物中的表达特性进行总结，对木质素降解机制及衍生芳烃代谢路径的研究进展进行综述。最后整理木质素降解微生物在秸秆肥料化技术中的应用进展，并探讨了微生物降解秸秆木质素的应用前景和未来的研究方向。     

细菌降解木质素的研究进展

木质素是自然界最丰富的芳香化合物,其分解与陆地上碳循环密切相关。提取木质纤维素中的葡萄糖使其转化成乙醇,是生产第二代生物能源的关键步骤。但是由于木质素是一种非常稳定的化合物,难以降解是实现生物乙醇转化的主要屏障,因此关于木质素的生物降解研究具有非常重要的意义。真菌降解木质素的研究已经深入的进行了多年,并取得丰富的成果,但是关于细菌降解木质素的研究还处在初级阶段。由于广泛的生长条件和良好的环境适应能力,细菌在木质素降解方面深受研究人员的关注。本文通过总结前人的研究成果,讨论了木质素的降解机制、代谢途径及细菌降解木质素的工业应用前景,同时还展望了分子生物学及生物信息学在木质素降解方面的应用前景。     

白腐菌木质素降解酶及其在木质素降解过程中的相互作用

木质素是一类不易降解的生物物质,在自然界中,白腐真菌对木质素的降解能力最强.白腐真菌降解木质素主要依靠分泌的三种酶:木质素过氧化物酶(Lip)、锰过氧化物酶(MnP)和漆酶(Lac).对白腐真菌分泌的三种木质素降解酶在性质、分布等方面进行了比较,系境地介绍三种木质素降解酶的催化作用,并阐述其在木质素降解过程中的相互作用.     

环境微生物介导的木质素代谢及其资源化利用研究进展

木质素是一种丰富的芳烃生物大分子聚合物,其分解代谢与地球元素循环和生物资源利用密切相关。但由于木质素结构的复杂性和无规则性导致其难以降解,使得木质素降解的研究成为全球碳循环和生物质资源利用研究的难点。近年来,来自不同环境的微生物陆续被发现具有木质素降解能力,并解析出参与木质素分解代谢的多种氧化还原酶。然而对木质素详细的代谢过程仍不十分清楚,因此,探究木质素降解酶系、作用机理和代谢网络是研究微生物代谢木质素机理的关键。本文综述环境中参与木质素降解的微生物,重点解析其木质素解聚酶系组成、分泌机制和木质素的代谢途径,并在此基础上阐明近年来木质素生物转化的最新研究进展,以期为今后环境微生物代谢木质素机理及其资源化利用的研究提供参考。     

木质素芳香族化合物降解菌Sphingobium sp. SYK-6的研究进展

木质素是木质的主要成分之一,在自然界中,高分子木质素被真菌的胞外酶分解成低分子芳香族化合物,然后土壤细菌将其完全降解为二氧化碳。由此可见,木质素的完全降解过程是真菌和细菌的共同作用。研究细菌的降解机制,一方面可以理解芳香族化合物在生态系中的碳素循环,另一方面可以为木质素的有效利用提供基因和酶工具,将可再生资源的木质素转化成高附加价值的工业产品。Sphingobium sp.SYK-6是1987年从造纸厂废水中以木质素中的联苯化合物(5,5’-脱氢联香草酸)作为唯一碳源分离出的木质素化合物降解菌。在长达25年以上的研究中我们阐明了一系列芳香族化合物的代谢途径,克隆了相关基因,2012年随着基因组测序的完成,整个降解功能的全貌展现出来。介绍内容:(1)基因组信息;(2)芳醚化合物代谢;(3)联苯化合物代谢;(4)阿魏酸代谢;(5)木质素化合物降解过程中四氢叶酸依赖型机制;(6)原儿茶酸4,5开环途径;(7)3-甲氧基没食子酸代谢的多样性;(8)应用研究。我们希望SYK-6菌株成为一个让人们理解木质素化合物降解的模式菌株。最后结合课题组现在的研究课题展望了木质素化合物的降解研究的发展方向。     

侧耳菌与粗毛栓菌在麦草培养基中产生木质纤维素降解酶的研究

草本植物,包括农作物秸杆的木质素主要是由松柏醇、芥子醇和对香豆醇的脱氢聚合物和对香豆酸组成[1,2],是结构复杂、稳定、多样的生物大分子物质.虽难于被一般微生物降解,但自然界中仍存在一些可降解木质素的微生物种类,白腐真菌是最重要的一类,它们通过分泌漆酶(Laccases,Lac)、木质素过氧化物酶(Lignin peroxidases,LiP)、锰过氧化物酶(Manganese-dependent peroxidases,MnP)、纤维素酶(Cellulas-es,Cel)和半纤维素酶(Hemicellulases,Hcel)等降解植物生物质.由于白腐菌在造纸工业中的生物制浆和纸浆生物漂白、环境保护等方面[4]有着很好的应用前景,因此倍受关注. 本研究选用在液体培养基中产酶能力强且产酶较快的白腐真菌侧耳sp2和粗毛栓菌[5]进行固体培养,研究它们产生木质纤维素降解酶类和降解植物生物质的能力.研究结果报道如下.     

木质纤维素降解菌及其绛解途径研究进展

文中综述了已发现的降解纤维素和术质素的主要的天然微生物种类、相关酶类,并着重介绍了白腐真菌和细菌降解木质素的过程。     

木质素降解酶及相关基因研究进展

生物质的高效综合利用已成为全球关注的热点问题。生物质的主要成分是木质素、纤维素和半纤维素,其利用的关键是如何去除木质素,从而提高纤维素和半纤维素的得率。其中利用真菌的生物预处理方法因条件温和、无二次污染等优点符合全球经济可持续发展需要,受到研究者的普遍关注。综述了近年国内外真菌分泌的主要木质素降解酶,包括木质素过氧化物酶(Li P)、锰过氧化物酶(Mn P)、漆酶(laccase)和多功能过氧化物酶(VP)的主要特点,总结了木质素降解相关酶的基因工程、基因组学的研究成果,并对其发展前景进行了展望。     

生物降解木质纤维素类生物质固废物的研究进展

自然界中的细菌、真菌和放线菌以及某些病毒都是能够降解某种纤维素的重要微生物，现代科技的发展使这种降解功能得以被发现。自然界的各种微生物在生物质固废能源转换和利用上具有十分重要的作用，不仅能够变废为宝，还能对生物质固废进行资源化利用。文章对生物质固废的相关处理技术、生物降解木质纤维素类生物质固废物的成果进行分析，总结了将微生物应用于生物质固废处理中的相关技术，并以物质纤维素固废为单位探讨分析相应的技术，旨在为我国的资源利用和绿色发展提供帮助与参考。     

低温秸秆降解微生物菌剂的研究进展

秸秆的成分主要有纤维素、半纤维素和木质素等,降解秸秆的微生物包括细菌、放线菌和真菌。低温秸秆降解微生物的选育方法有直接从自然界中筛选、诱变育种、原生质体融合育种、基因工程育种等。目前,筛选获得的低温秸秆降解菌株的数量有限,降解秸秆的能力不高,低温条件下菌株的降解机理都需要进一步的研究。综述了低温条件下秸秆降解微生物菌剂的研究进展。     

Lignin degradation by microorganisms: A review

Lignin is an abundant plant-based biopolymer that has found applications in a variety of industries from construction to bioethanol production. This recalcitrant branched polymer is naturally degraded by many different species of microorganisms, including fungi and bacteria. These microbial lignin degradation mechanisms provide a host of possibilities to overcome the challenges of using harmful chemicals to degrade lignin biowaste in many industries. The classes and mechanisms of different microbial lignin degradation options available in nature form the primary focus of the present review. This review first discusses the chemical building blocks of lignin and the industrial sources and applications of this multifaceted polymer. The review further places emphasis on the degradation of lignin by natural means, discussing in detail the lignin degradation activities of various fungal and bacterial species. The lignin-degrading enzymes produced by various microbial species, specifically white-rot fungi, brown-rot fungi, and bacteria, are described. In the end, possible directions for future lignin biodegradation applications and research investigations have been provided.     

Biotransformation of lignin: Mechanisms,applications and future work

As one of the most abundant polymers in biosphere, lignin has attracted extensive attention as a kind of promising feedstock for biofuel and bio-based products. However, the utilization of lignin presents various challenges in that its complex composition and structure and high resistance to degradation. Lignin conversion through biological platform harnesses the catalytic power of microorganisms to decompose complex lignin molecules and obtain value-added products through biosynthesis. Given the heterogeneity of lignin, various microbial metabolic pathways are involved in lignin bioconversion processes, which has been characterized in extensive research work. With different types of lignin substrates (e.g., model compounds, technical lignin, and lignocellulosic biomass), several bacterial and fungal species have been proved to own lignin-degrading abilities and accumulate microbial products (e.g., lipid and polyhydroxyalkanoates), while the lignin conversion efficiencies are still relatively low. Genetic and metabolic strategies have been developed to enhance lignin biodegradation by reprogramming microbial metabolism, and diverse products, such as vanillin and dicarboxylic acids were also produced from lignin. This article aims at presenting a comprehensive review on lignin bioconversion including lignin degradation mechanisms, metabolic pathways, and applications for the production of value-added bioproducts. Advanced techniques on genetic and metabolic engineering are also covered in the recent development of biological platforms for lignin utilization. To conclude this article, the existing challenges for efficient lignin bioprocessing are analyzed and possible directions for future work are proposed.     

Recent Developments in Using Advanced Sequencing Technologies for the Genomic Studies of Lignin and Cellulose Degrading Microorganisms

Lignin is a complex polyphenyl aromatic compound which exists in tight associations with cellulose and hemicellulose to form plant primary and secondary cell wall. Lignocellulose is an abundant renewable biomaterial present on the earth. It has gained much attention in the scientific community in recent years because of its potential applications in bio-based industries. Microbial degradation of lignocellulose polymers was well studied in wood decaying fungi. Based on the plant materials they degrade these fungi were classified as white rot, brown rot and soft rot. However, some groups of bacteria belonging to the actinomycetes, α-proteobacteria and β-proteobacteria were also found to be efficient in degrading lignocellulosic biomass but not well understood unlike the fungi. In this review we focus on recent advancements deployed for finding and understanding the lignocellulose degradation by microorganisms. Conventional molecular methods like sequencing 16s rRNA and Inter Transcribed Spacer (ITS) regions were used for identification and classification of microbes. Recent progression in genomics mainly next generation sequencing technologies made the whole genome sequencing of microbes possible in a great ease. The whole genome sequence studies reveals high quality information about genes and canonical pathways involved in the lignin and other cell wall components degradation.     

A field of dreams: Lignin valorization into chemicals,materials, fuels,and health-care products

Lignin is one of the most abundant renewable resources on earth and is readily produced as a sidestream during biomass fractioning. So far, these large quantities of lignin have been severely underutilized, thereby wasting this valuable renewable. Recent technological advances in lignin recovery, breakdown, and conversion have now started forming the first sustainable value chains to take advantage of lignin. Microbial cell factories, inspired by nature’s miscellaneous set of lignin-degrading microbes, are at the heart of these novel processes. Recent success stories in which the enzymes and pathways of these microbes were harnessed for biobased production from lignin hold great promise for a sustainable upgrading of this renewable polymer into value-added compounds.     

Microbial degradation of lignin

The transformations of lignin that occur during its biodegradation are complex and incompletely understood. Certain fungi of the white-rot group, and possibly other fungi and bacteria, completely decompose lignin to carbon dioxide and water. Other fungi and bacteria apparently degrade lignin incompletely. Differences in lignin-degrading abilities observed for different organisms may result from differences in the completeness of their ligninolytic enzyme systems. Not all lignin components may be attacked by a particular organism. Alternatively, different organisms may differ in their basic mechanisms of attack on lignin. The basic pathways of lignin degradation have been elucidated only for certain representatives of the white-and brown-rot fungi. Although it is known that each of the principal structural components of lignin is attacked by other fungi and bacteria, the biochemistry of that attack has not been elucidated. Work with low molecular weight lignin models has provided only limited information on possible pathways of lignin degradation by microorganisms. There is little evidence to suggest a correlation between abilities to degrade single-ring aromatic or lignin model compounds and the ability to degrade polymeric lignin. More evidence has come from analysis of spent culture media for lignin breakdown products and from comparative chemical analyses of sound lignins versus decayed lignin residues. Accumulated evidence with the most thoroughly studied white-rot fungi suggests that with these fungi lignin degradation proceeds by way of extracellular mixed-function oxygenases and dioxygenases, which catalyse demethylations, hydroxylations and ring-fission reactions within a largely intact polymer, concomitant with some release of low molecular weight lignin fragments. There are also apparent relationships between lignin, carbohydrate and nitrogen metabolism for some organisms, but the relationships may vary from one organism to another. Although research is now mostly at a basic level, industrial applications may result from lignin degradation research. Considerable potential exists for the development of bioconversions which might produce low molecular weight chemicals from waste lignins, and thereby reduce our dependence on petroleum as a source of these chemicals. Alternatively, such bioconversions might produce chemically altered forms of polymeric lignin that may be valuable industrially.     

远海50个站位沉积物中潜在木质素降解菌的分离与鉴定

【背景】微生物在海洋木质素的降解过程中发挥了至关重要的作用,然而来源海洋环境的木质素降解菌的相关研究报道却很少。【目的】从远海沉积物环境分离潜在的木质素降解菌,为木质素的可再生化学物质转化提供菌种资源。【方法】利用以木质素为唯一碳源的培养基,对50个远海沉积物样品中的木质素降解菌进行富集培养与纯化,并利用含苯胺蓝的脱色培养基筛选潜在木质素降解菌,继而通过16S rRNA基因测序与序列比对初步确定潜在木质素降解菌的分类地位。【结果】从50个沉积物样品中共分离获得菌株283株,其中潜在木质素降解菌263株,它们隶属于α-变形杆菌纲(Alphapreobacteria)、γ-变形杆菌纲(Gamaproteobacteria)、芽孢杆菌纲(Bacilli)、放线杆菌纲(Actinobacteria)和黄杆菌纲(Flavobacteriia)中的32个属。【结论】远海沉积物环境中蕴含着丰富的木质素降解菌,为海洋生物资源的开发利用提供了新的参考。     

Characterization of trapped lignin-degrading microbes in tropical forest soil

Lignin is often the most difficult portion of plant biomass to degrade, with fungi generally thought to dominate during late stage decomposition. Lignin in feedstock plant material represents a barrier to more efficient plant biomass conversion and can also hinder enzymatic access to cellulose, which is critical for biofuels production. Tropical rain forest soils in Puerto Rico are characterized by frequent anoxic conditions and fluctuating redox, suggesting the presence of lignin-degrading organisms and mechanisms that are different from known fungal decomposers and oxygen-dependent enzyme activities. We explored microbial lignin-degraders by burying bio-traps containing lignin-amended and unamended biosep beads in the soil for 1, 4, 13 and 30 weeks. At each time point, phenol oxidase and peroxidase enzyme activity was found to be elevated in the lignin-amended versus the unamended beads, while cellulolytic enzyme activities were significantly depressed in lignin-amended beads. Quantitative PCR of bacterial communities showed more bacterial colonization in the lignin-amended compared to the unamended beads after one and four weeks, suggesting that the lignin supported increased bacterial abundance. The microbial community was analyzed by small subunit 16S ribosomal RNA genes using microarray (PhyloChip) and by high-throughput amplicon pyrosequencing based on universal primers targeting bacterial, archaeal, and eukaryotic communities. Community trends were significantly affected by time and the presence of lignin on the beads. Lignin-amended beads have higher relative abundances of representatives from the phyla Actinobacteria, Firmicutes, Acidobacteria and Proteobacteria compared to unamended beads. This study suggests that in low and fluctuating redox soils, bacteria could play a role in anaerobic lignin decomposition.     

A comparison between the homocyclic aromatic metabolic pathways from plant-derived compounds by bacteria and fungi

Aromatic compounds derived from lignin are of great interest for renewable biotechnical applications. They can serve in many industries e.g. as biochemical building blocks for bioplastics or biofuels, or as antioxidants, flavor agents or food preservatives. In nature, lignin is degraded by microorganisms, which results in the release of homocyclic aromatic compounds. Homocyclic aromatic compounds can also be linked to polysaccharides, tannins and even found freely in plant biomass. As these compounds are often toxic to microbes already at low concentrations, they need to be degraded or converted to less toxic forms. Prior to ring cleavage, the plant- and lignin-derived aromatic compounds are converted to seven central ring-fission intermediates, i.e. catechol, protocatechuic acid, hydroxyquinol, hydroquinone, gentisic acid, gallic acid and pyrogallol through complex aromatic metabolic pathways and used as energy source in the tricarboxylic acid cycle. Over the decades, bacterial aromatic metabolism has been described in great detail. However, the studies on fungal aromatic pathways are scattered over different pathways and species, complicating a comprehensive view of fungal aromatic metabolism. In this review, we depicted the similarities and differences of the reported aromatic metabolic pathways in fungi and bacteria. Although both microorganisms share the main conversion routes, many alternative pathways are observed in fungi. Understanding the microbial aromatic metabolic pathways could lead to metabolic engineering for strain improvement and promote valorization of lignin and related aromatic compounds.