堆肥化中木质素的生物降解

木质素是堆肥化原料中一种重要的限速高聚物,其有效降解对堆肥化速度、堆肥质量有重要作用.综述了堆肥化中木质素生物降解的研究进展,包括堆肥化中降解木质素的微生物种类及其降解过程和机理,以及木质素的降解与堆肥化中腐殖质形成的关系.     

物料预处理对堆肥减量化、腐殖化和稳定化的影响及其微生物机制

堆肥是有机固体废弃物处理与资源化的主要途径之一,包括矿化和腐殖化两个过程,且都和微生物活动有关。矿化过程会产生二氧化碳(CO2)等温室气体,是一个典型的温室气体释放过程。腐殖化过程则会产生稳定的腐殖质,则是优良的土壤改良剂。在堆肥稳定化的前提下,如何有效减少堆肥过程中的CO2释放,强化堆肥的腐殖化过程,增加作为优良土壤改良剂的腐殖质产量,对于发展低碳化堆肥技术,实现堆肥的资源化利用具有重要意义。本文选取水稻秸秆和餐厨垃圾作为堆肥原料,研究不同预处理对堆肥过程中矿化和腐殖化过程的影响,并探讨了不同预处理影响矿化和腐殖化过程的微生物机理。结果表明堆料加热预处理后,堆肥的矿化作用被明显削弱,总碳(TC)减量率仅为23.4%,并且最后形成了可观产量的稳定腐殖质(每kg堆料70 d后腐殖质含量为22.09 g±0.35 g,腐殖化系数达2.0),因此加热预处理后的堆肥过程在保证稳定腐殖质的产量前提下更低碳化。预处理通过影响堆料的性质和初始状态下堆料中微生物的种类和数量从而影响堆肥的矿化和腐殖化过程。活性微生物量与脱氢酶活性是矿化过程的主要决定因素,而多酚氧化酶活性主要影响堆肥的稳定腐殖化过程。     

真菌降解木质素研究进展及在好氧堆肥中的研究展望

综述了近十年来真菌降解木质素的研究进展,包括木质素的存在与结构,真菌降解木质素生物学、酶系及作用机理、生理学以及在环境工程中应用方面的研究进展,并对好氧堆肥处理城市垃圾中木质素生物降解的研究作了展望 。     

微生物法降解木质素的研究进展

生物质是代替石化资源生产能源和化学品的关键资源,木质素作为植物细胞壁的主要成分已经在很多行业中得到了广泛的应用。然而,由于木质素结构复杂且难以降解,成为生物质资源利用的最大障碍,因此,去除或者降解木质素是利用细胞壁中其他成分的关键步骤。许多行业使用有害化学物质降解木质素,严重危害了生态环境,自然界中木质素经常被包括真菌和细菌在内的微生物降解,因此,研究微生物降解木质素的机制为解决这一问题提供了可能性。本文讨论了木质素的化学组成成分,重点讨论了自然界降解木质素的微生物种类及其降解机制,包括各种真菌和细菌的木质素降解活性,描述了由各种微生物特别是白腐真菌、褐腐真菌和细菌产生的木质素降解酶,并展望了今后木质素生物降解的研究和应用的可能方向。     

基于环境微生物资源库快速构建易腐垃圾高效堆肥复合菌剂

【目的】针对易腐垃圾成分快速构建一种高效堆肥复合菌剂。【方法】研究以"中国科学院战略生物资源服务网络计划"支持建设的微生物资源库为基础,根据菌株产酶活功能信息,针对易腐垃圾有机成分定向挑选出高产酶(蛋白酶、淀粉酶、纤维素酶、木质素酶、脂肪酶)菌株,通过测定菌株间拮抗作用,构建了一种高效堆肥复合菌剂CM菌剂。进行了菌剂发酵条件优化和易腐垃圾堆肥应用研究。【结果】CM菌剂最适发酵条件为接种7%的种子液于红糖培养基中,30°C下培养48h,此时菌剂产生蛋白酶、淀粉酶、纤维素酶、木质素酶、脂肪酶酶活分别为181.76、52.92、1.57、12.81、9.94 U/mL。堆肥结果表明CM菌剂堆肥产品pH、含水率、有机质含量均满足生物有机肥标准(NY884-2012)。CM菌剂堆肥过程中最高温度为63.5°C,高温期9–12 d,30 d后含水率为28.7%,有机质降解率为30.4%,C/N为8.93,与商品菌剂BM相比堆肥效果更好,可使堆体高温期延长4–7d、含水率降低3.8%,可加快易腐垃圾中有机质降解,降解率提升6.2%,并具有更强的固氮能力,缩短腐熟时间,提升堆肥产品品质。【结论】基于环境微生物资源库功能信息构建堆肥复合菌剂是一种快速有效的菌剂构建方法。     

木质素的微生物降解机制

研究微生物降解木质素的反应机理,可以从根本上解释微生物或酶对木质素的作用过程,对提高木质素降解效率,治理环境污染等具有非常重要的意义。从木质素结构的差异出发,总结了近年来研究木质素微生物降解机制所采用的主要模型化合物、研究方法,概述了微生物对木质素的三大作用机理:侧链氧化、去甲基化和芳香环断裂,以及参与这三个反应的主要微生物。     

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。     

一株木质素降解菌高温驯化及酶学性质的测定

【背景】高温引起的微生物活性降低是限制园林绿化废弃物堆肥过程中木质素降解的主要因素。【目的】驯化一株木质素降解菌芽孢杆菌NO.2,提高其在高温下的微生物活性,探究其生长情况及酶学特性。【方法】采用温度梯度方法驯化菌株,以菌株生长曲线、酶活力、木质素降解率为评价指标,探究驯化前后菌株间差异,以及驯化后菌株所产木质素降解酶的酶促反应温度和pH范围。【结果】与原菌株相比,驯化后菌株在60℃培养时最大生物量间差异不显著;漆酶(laccase,Lac)、锰过氧化物酶(manganeseperoxidase,MnP)和木质素过氧化物酶(ligninperoxidase,LiP)酶活力得到进一步提高,分别提高了30.75%、35.98%和29.62%,木质素降解率提高60.52%。酶学性质研究表明,驯化后菌株所产Lac、MnP和LiP在20-60℃、pH 3.0-9.0范围内酶活力均较高,而且具有较好的稳定性,稳定性依次为Lac>LiP>MnP。【结论】温度梯度驯化方法可有效提高微生物对高温环境的适应性,扩大木质素降解酶的酶促反应温度和pH范围,在进一步自主研制专用降解园林废弃物微生物菌...     

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。     

C/N驱动优势细菌菌群变化影响堆肥碳氮损失和腐殖质合成

为了探明C/N如何驱动堆肥过程中优势细菌菌群的变化而影响碳氮损失和腐殖质合成,设置3个C/N处理(20∶1、25∶1和30∶1),以羊粪和玉米秸秆为原料进行堆肥试验。结果表明: 与20∶1处理相比,30∶1和25∶1处理堆肥的碳、氮损失分别降低了33.5%、18.9%和23.6%、10.8%。优势细菌菌群、碳氮损失及有机碳组分的冗余分析表明,高C/N提高了堆肥中固氮细菌的种类和丰度,降低了反硝化细菌的种类和丰度,减少了堆肥过程中的碳氮损失;高C/N促进了木质纤维素类降解菌的生长繁殖,促进了富里酸和胡敏素降解而合成更多胡敏酸,提高了堆肥腐殖化程度。可见,C/N可通过影响堆肥中关键优势细菌菌群而影响堆肥过程和堆肥质量,调节堆肥原料C/N可以调控堆肥中碳氮损失和腐殖质的合成,从而提高堆肥质量并减少堆肥的二次环境污染。     

Microbial indicators for sawdust and bark compost stability and humification processes

To what extent some microbial index ratios are suitable for use as early criteria for the level of compost stability during aerobic composting of coniferous sawdust and bark at mesophilic conditions was studied. Evolution of the specific respiration activity (CO₂-C/biomass C) and the ratios between some groups of microorganisms were followed as a function of composting time. The specific respiration activity was found to be an early and most reliable indicator of compost stability. The peroxidase and polyphenoloxidase enzyme activity during composting, as well as the composition of newly-formed humus substances were studied. The duration of composting increased the quality of newly-formed humus substances (C_h.a.:C_f.a ratio; Ca-complexed humic acid and resistance of organo-mineral complexes). The quality of humus substances could be used to assess compost stability. However, the results can be applied only under defined conditions.     

Effect of bioaugmentation and nitrogen supplementation on composting of paddy straw

A composting experiment was conducted to evaluate the effect of a hyperlignocellulolytic fungal consortium and different nitrogen amendments on paddy straw composting in terms of changes in physicochemical and biological parameters. A fungal consortium comprising four lignocellulolytic mesophilic fungal cultures was used as inoculum for bioaugmentation of paddy straw in perforated pits. The comparative effect of farmyard manure (FYM), soybean trash, poultry litter and urea on the composting process was evaluated at monthly intervals in terms of physicochemical (pH, EC, available P, C:N ratio and humus content) and biological (enzymatic and microbial activity) parameters. The compost prepared from bioaugmented paddy straw composting mixture, with poultry manure as nitrogen supplement attained desirable C:N ratio in 1 month and displayed least phytotoxicity levels along with higher production of β-1,4-Exoglucanase. The combined activity of the autochthonous composting microbiota as well as the externally applied fungal inoculum accelerated the composting process of paddy straw. Supplementation of paddy straw with poultry manure in 8:1 ratio was identified as the best treatment to hasten the composting process. This study highlights the importance of application of fungal inoculum and an appropriate N-amendment such as poultry manure for preparation of compost using a substrate having high C:N ratio, such as paddy straw.     

翻堆频率对堆肥减量化、腐殖化和稳定化的影响

好氧堆肥是有机固体废弃物处理处置的有效手段之一,堆肥还田也是贫瘠土壤改良的常用措施。但好氧堆肥是一个典型的CO_2等温室气体的释放过程。如何减少堆肥过程中的CO_2释放,强化堆肥的腐殖化过程对于实现有机固体废弃物的低碳化堆肥、提高作为优良土壤改良剂的腐殖质产量具有重要意义。本文选取农林秸秆和餐厨垃圾作为堆肥原料,研究不同翻堆频率对堆肥过程中的物料减量化、腐殖化和稳定化的影响,以期发现一个较低碳的堆肥工艺,并从微生物角度初步探索了其影响机制。研究结果显示,不同的翻堆频率(分别为每2 d、4 d和6 d翻堆一次),堆料的减量化和腐殖化程度有一定差异,翻堆频率为4 d的堆肥工艺物料减量率最高为50.5%,但碳减量率最低为77.4%;而翻堆频率为2 d的堆肥工艺腐殖质产量最高;3种堆肥工艺经62 d堆肥都达到了腐熟程度,翻堆频率为4 d的堆肥工艺腐熟化程度最高。不同的翻堆频率可能通过影响堆肥过程中堆料的温度、含氧量等因素从而改变堆料中活性微生物量、种类和生物酶活性,进而影响堆料的矿化和腐殖化进程。     

Are Basidiomycete Laccase Gene Abundance and Composition Related to Reduced Lignolytic Activity Under Elevated Atmospheric NO<Subscript>3</Subscript><Superscript>−</Superscript> Deposition in a Northern Hardwood Forest?

Anthropogenic release of biologically available N has increased atmospheric N deposition in forest ecosystems, which may slow decomposition by reducing the lignolytic activity of white-rot fungi. We investigated the potential for atmospheric N deposition to reduce the abundance and alter the composition of lignolytic basidiomycetes in a regional network of four northern hardwood forest stands receiving experimental NO₃ ⁻ deposition (30 kg NO₃ ⁻−N ha⁻¹ year⁻¹) for a decade. To estimate the abundance of basidiomycetes with lignolytic potential, we used PCR primers targeting laccase (polyphenol oxidase) and quantitative fluorescence PCR to estimate gene copy number. Natural variation in laccase gene size permitted use of length heterogeneity PCR to profile basidiomycete community composition across two sampling dates in forest floor and mineral soil. Although past work has identified significant and consistent negative effects of NO₃ ⁻ deposition on lignolytic enzyme activity, microbial biomass, soil respiration, and decomposition rate, we found no consistent effect of NO₃ ⁻ deposition on basidiomycete laccase gene abundance or community profile. Rather, laccase abundance under NO₃ ⁻ deposition was lower (−52%), higher (+223%), or unchanged, depending on stand. Only a single stand exhibited a significant change in basidiomycete laccase gene profile. Basidiomycete laccase genes occurring in mineral soil were a subset of the genes observed in the forest floor. Moreover, significant effects on laccase abundance were confined to the forest floor, suggesting that species composition plays some role in determining how lignolytic basidiomycetes are affected by N deposition. Community profiles differed between July and October sampling dates, and basidiomycete communities sampled in October had lower laccase gene abundance in the forest floor, but higher laccase abundance in mineral soil. Although experimental N deposition significantly suppresses lignolytic activity in these forests, this change is not related to the abundance or community composition of basidiomycete fungi with laccase genes. Understanding the expression of laccases and other lignolytic enzymes by basidiomycete fungi and other lignin-decaying organisms appears to hold promise for explaining the consistent decline in lignolytic activity elicited by experimental N deposition. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Efficient,environmentally-friendly and specific valorization of lignin: promising role of non-radical lignolytic enzymes

Lignin is the second most abundant bio-resource in nature. It is increasingly important to convert lignin into high value-added chemicals to accelerate the development of the lignocellulose biorefinery. Over the past several decades, physical and chemical methods have been widely explored to degrade lignin and convert it into valuable chemicals. Unfortunately, these developments have lagged because of several difficulties, of which high energy consumption and non-specific cleavage of chemical bonds in lignin remain the greatest challenges. A large number of enzymes have been discovered for lignin degradation and these are classified as radical lignolytic enzymes and non-radical lignolytic enzymes. Radical lignolytic enzymes, including laccases, lignin peroxidases, manganese peroxidases and versatile peroxidases, are radical-based bio-catalysts, which degrade lignins through non-specific cleavage of chemical bonds but can also catalyze the radical-based re-polymerization of lignin fragments. In contrast, non-radical lignolytic enzymes selectively cleave chemical bonds in lignin and lignin model compounds and, thus, show promise for use in the preparation of high value-added chemicals. In this mini-review, recent developments on non-radical lignolytic enzymes are discussed. These include recently discovered non-radical lignolytic enzymes, their metabolic pathways for lignin conversion, their recent application in the lignin biorefinery, and the combination of bio-catalysts with physical/chemical methods for industrial development of the lignin refinery.     

Survival of Human Pathogens in Composted Sewage

Studies were conducted to assess the effectiveness of an aerobic composter in destroying pathogens that may possibly be present in raw sewage sludge. Experiments conducted in this study were designed to determine whether or not selected indicator organisms (i.e., Salmonella newport, poliovirus type 1, Ascaris lumbricoides ova, and Candida albicans) could survive the composting process. The results of the assay showed that after 43 hr of composting, no viable indicator organisms could be detected. The poliovirus type I was the most sensitive, being inactivated within the first hour, whereas C. albicans was the most resistant, requiring more than 28 hr of composting for its inactivation. The data from this study indicated that aerobic composting of sewage sludge would destroy the indicator pathogens when a temperature of 60 to 70 C is maintained for a period of 3 days.     

Effects of low pH and humus on the survivorship, growth and feeding of Gammarus pulex (L.) (Amphipoda)

SUMMARY. 1. The toxicity to the freshwater amphipod Gammarus pulex (L.) of soft water (pH6.0) with three concentrations of XAD-extracted aquatic humus was treated in the laboratory.
2. Exposure to pH6.0 water without humus added resulted in 92% mortality after 3 weeks. Humus, added at concentrations of 7 and 20 g Cm ³, decreased the mortality to 80% and 64% in the same period.
3. Surviving animals kept at pH6.0 had a lower growth rate, lower food conversion efficiency and higher body water content than animals kept at pH 7.3.
4. Humus had no significant effects on growth, food conversion or body composition of G. pulex kept at pH 6.0. However, there was a tendency for growth and food conversion to increase, and for body water content to decrease with increased humus concentration. The effects of humus on growth and food conversion of G. pulex observed in this study do not support the contention that humus acts as a free coupler to lower metabolic efficiency. On the contrary, humus tended to benefit food conversion.
5. It is suggested that low concentrations of humus can be directly beneficial to organisms in acidified water in ways other than by complexing toxic metals.     

Effects of struvite formation and nitratation promotion on nitrogenous emissions such as NH3, N2O and NO during swine manure composting

To reduce nitrogenous emissions from composting, two different countermeasures were applied simultaneously in swine manure composting. One was forming struvite by adding Mg and P at the start of composting, and the other was to promote nitratation (nitrite being oxidized nitrate) by adding nitrite-oxidizing bacteria after the thermophilic phase of composting. In the laboratory- and mid-scale composting experiments, 25-43% of NH3, 52-80% of N2O and 96-99% of NO emissions were reduced. From the nitrogen balance, it was revealed that the struvite formation reduced not only NH3, but also other nitrogenous emissions except N2O. The amount of total nitrogen losses was reduced by 60% by the two combined countermeasures, against 51% by the struvite formation alone. However, the nitratation promotion dissolved struvite crystals due to the pH decline, diminishing the effect of struvite as a slow-release fertilizer.     

一株分离自绿化废弃物堆肥枯草芽孢杆菌的常压室温等离子诱变及其发酵条件

【背景】为了提高堆肥降解有机废弃物的效率,高效堆肥菌剂成为了研究热点,其中以真菌应用的研究为多,但真菌也有对氧气和底物敏感等缺点,细菌对堆肥的作用开始被研究。本实验室以羧甲基纤维素钠(CMC-Na)为底物,从绿化废弃物堆肥中筛选得到枯草芽孢杆菌(Bacillussubtilis,B.subtilis) BL03,它具有较好的纤维素分解能力,能提高绿化废弃物堆肥中纤维素降解和腐殖质合成的速度。【目的】进一步提高B.subtilisBL03的纤维素酶生产能力。【方法】利用常压室温等离子(Atmospheric and room temperature plasma,ARTP)诱变BL03菌,通过CMC-刚果红固体培养基观察水解透明圈,以及液体发酵后检测酶活力的方法进行3轮筛选;通过连续多代培养观察突变株的遗传稳定性;通过梯度温度、p H培养研究突变株发酵的最适生长温度、培养基初始pH;利用正交设计方法研究适合突变株发酵培养的工业级原料配方。【结果】筛选到2株正突变株,酶活力分别提高了69%和72%;连续10代培养稳定,验证了突变株的遗传稳定性;其中酶活力最高的突变株BLA3890最适培养温度为37°C、培养基初始pH为5.0-6.5,研究得到较经济的发酵培养基配方。【结论】ARTP诱变B. subtilis BL03后得到的突变株BLA1973和BLA3890在绿化废弃物堆肥或其他纤维素降解行业具有进一步研究和应用的价值。     

Composting: a potentially safe process for disposal of genetically modified organisms

The widespread use of genetically modified organisms (GMOs) may result in the release of GMOs into the environment. The potential risks regarding their use and implementation of disposal methods, especially the possibility of novel genes from GMOs being transferred to natural organisms, need to be evaluated and better understood. There is an increasingly accepted public view that GMO products introduced into the environment should be degradable and should disappear after a limited period of time. Due to the risk of possible horizontal gene transfer, disposal methods for GMOs need to address destruction of both the organism and the genetic material. During the last two decades, we have developed a greater understanding of the biochemical, microbiological and molecular concepts of the composting process, such that maximum decomposition may be achieved in the shortest time with minimal negative impacts to the environment. The conditions created in a properly managed composting process environment may help in destroying GMOs and their genes, thereby reducing the risk of the spread of genetic material. When considering composting as a potential method for the disposal of GMOs, the establishment of controlled conditions providing an essentially homogenous environment appears to be an important requirement. An evaluation of composting as a safe option for disposal of GMOs is provided in this review.