生物垃圾好氧处理中的纤维素降解菌生长规律研究

目的:研究了蔬菜垃圾好氧处理过程中,纤维素降解菌和半纤维素降解菌(细菌和真菌),纤维素酶活和半纤维素酶活,和有机物降解之间的变化规律。方法:用添加纤维素和半纤维素的牛肉膏蛋白胨培养基和查式培养基,分别培养计数纤维素降解细菌、真菌和半纤维素降解细菌、真菌;马福炉灼烧测有机物含量。结果:好氧处理的初始阶段中,前4d有机物日均降解率5.2%,后3d日均降解率2.2%。结论:半纤维素降解菌的数量比纤维素降解菌的多,半纤维素酶活力,也高于纤维素酶活力;微生物的变化情况为前6d产两种酶的微生物主要有细菌和真菌;从第6d开始真菌快速生长;至第7d真菌纤维素酶和半纤维素酶活力显著升高。     

5种优势腐生真菌降解华山松针叶的酶活测定

【目的】分析腐生真菌降解华山松落针过程中各种酶活性变化与酶间关系,探索真菌对华山松落针的降解能力。【方法】通过形态学观察和转录间隔区ITS序列分析,鉴定分离自华山松凋落物的5种优势菌株。以此为供试菌株,华山松落针为底物,通过发酵纯培养方法,测定底物总有机物质(Total organic matter,TOM)质量损失率及在发酵过程中产生的内切葡聚糖酶(Endoglucanase,EG)、木聚糖酶(Xylanase,Xyl)、木质素过氧化物酶(Lignin peroxidase,Li P)、锰过氧化物酶(Manganese peroxidase,Mn P)和酸性磷酸酶(Acid phosphatase,AP)活性。【结果】5种菌株分别为Mucor sp.、Pestalotiopsis sp.、Allantophomopsis sp.、Phoma sp.和Hypocrea sp.。5种菌引起的针叶TOM质量损失率在6.63%-15.77%之间,Pestalotiopsis sp.具有最高的AP酶活性,且EG酶、Xyl酶和Li P酶3种酶活性较高。Allantophomopsis sp.的Li P酶活性最高,并具有很高的Mn P酶活。Hypocrea sp.分泌的EG酶、Xyl酶活性低,但能产生Li P酶且有较高的AP酶活。相关性分析表明菌株分泌的AP酶活性与TOM质量损失率负相关,EG酶、Xyl酶及AP酶3种酶之间存在协同作用,特别是EG与AP之间。【结论】5种腐生真菌对华山松针叶均有降解作用,降解能力:Pestalotiopsis sp.>Allantophomopsis sp.>Hypocrea sp.>Phoma sp.>Mucor sp.。酶活大小及酶协同作用共同影响针叶降解,Pestalotiopsis sp.、Allantophomopsis sp.和Hypocrea sp.能产生木质纤维素降解酶并能引起较高的质量损失率,因此这3株菌为木质纤维素降解真菌。     

木聚糖酶对不同品种小麦戊聚糖降解的影响作用研究

以黑曲木霉C2-56菌株所产木聚糖酶为材料,研究木聚糖酶对不同小麦品种籽粒戊聚糖降解的影响作用。选取4个代表性小麦品种随机区组、重复设计,测定酶解前、后的戊聚糖含量,并对数据进行统计学处理。木聚糖酶降解不同小麦品种戊聚糖的差异不显著。本研究为合理利用小麦资源、开展专用小麦育种等方面提供理论参考。     

不同碳源和氮源对金针菇降解木质纤维素酶活性的影响

以3株栽培的金针菇Flammulina velutipes为材料,研究它们在玉米芯和棉子壳以及不同碳源、氮源培养条件下纤维素、半纤维素和木质素降解酶活性的规律。结果表明,不同金针菇菌株的羧甲基纤维素酶、木聚糖酶和漆酶活力显著不同（P<0.001）,同时,培养条件对羧甲基纤维素酶、木聚糖酶和漆酶的活力都有显著影响（P<0.001）。在简单碳源存在的条件下,金针菇的羧甲基纤维素酶和木聚糖酶活力远远低于复杂碳源培养基（P<0.05）。全营养培养基上生长的金针菇的羧甲基纤维素酶和木聚糖酶活力低于缺乏碳源和氮源的培养基（P<0.05）。漆酶活力在无简单氮源培养基上低于全培养基（P<0.05）和无葡萄糖培养基（P<0.05）,即复杂碳源和氮源培养基上的漆酶活力低于简单碳源和氮源培养基（P<0.05）。     

瘤胃微生物在有机废物处理中的应用研究

综述了瘤胃微生物在处理农业残余废物、城市有机垃圾和一些有毒物质方面的研究情况,并对影响其降解的环境条件、工艺条件和反应促进因素做了介绍,认为结合现代厌氧消化技术和瘤胃发酵技术,瘤胃微生物可以在有机废物处理中发挥较大作用.     

不同酸中和对垃圾渗滤液厌氧生物处理的影响

不同时间垃圾填埋场渗滤液用石灰絮凝、吹脱后分别经磷酸、盐酸、硫酸调pH到7,在35℃条件下进行厌氧处理。试验表明,经磷酸或盐酸中和的早期垃圾渗滤液易于厌氧生物处理,38d后其COD、BOD5、VFA（挥发性脂肪酸）都有大幅度的降低。盐酸中和的早期垃圾渗滤液其厌氧产甲烷性能良好;但磷酸中和的其产甲烷性能被完全抑制。硫酸中和的早期垃圾渗滤液在反应过程中产生大量的硫化物,最高浓度达到1.241mg／L,对厌氧处理产生了严重的抑制,但是在第38d硫化物浓度达到最高后抑制作用慢慢减弱,其COD、BOD5、VFA开始迅速下降。晚期垃圾渗滤液经磷酸、盐酸、硫酸中和后经过54d的反应,其COD、BOD5、VFA均没有明显降低。     

异养硝化⁃好氧反硝化复合菌剂在垃圾渗滤液处理中的应用

垃圾渗滤液中往往含有高浓度的有机物、氨氮等污染物。异养硝化-好氧反硝化型微生物能在脱氮的同时去除部分有机物,但目前对于相关混合菌剂直接应用于垃圾渗滤液处理的研究较少。从垃圾渗滤液中筛选出6株异养硝化-好氧反硝化菌株并组配成复合菌剂F6,探究菌剂在垃圾渗滤液中的脱氮效果。分别以单一菌株和复合菌剂F6为投放原料;以不同碳氮比、活性炭浓度、转速以及微量元素浓度为影响因素,研究复合菌剂对于垃圾渗滤液中氨氮(NH{}_{\mathrm{4}}^{+}-N)、总氮(total nitrogen, TN)、化学需氧量(chemical oxygen demand, COD)的去除性能。结果表明,提高碳氮比和微量元素浓度能够促进复合菌剂F6的降解效果;当接种量为10%、碳氮比为15%时,F6对NH{}_{\mathrm{4}}^{+}-N、TN、COD的去除率分别为74.69%、89.23%和83.50%,与不添加活性炭的处理相比,分别提高了约18.46%、20.97%和7.98%。复合异养硝化-好氧反硝化菌剂F6对高氨氮垃圾渗滤液去除NH{}_{\mathrm{4}}^{+}-N、TN、COD等方面具有良好的应用前景。     

低温微生物在污水处理中的应用分析

低温微生物作为极端环境微生物研究的一大热点,自20世纪70年代以来,其研究应用就得到了越来越广泛的关注。低温微生物凭借一系列耐冷机制,可有效抵御低温对其细胞生存生长的胁迫,保障其在低温环境下仍能够生存繁殖。文章首先阐述了低温微生物的相关内涵及其适冷机制,然后探讨了低温微生物在污水处理中的实际应用,以期为低温微生物在实际生产生活中的应用提供参考。     

外接堆肥微生物在餐厨废弃物好氧堆肥中的应用

微生物是堆肥过程的主要驱动力,其群落在堆肥过程中不断演替,进而影响好氧发酵进程和堆肥的品质。大量研究表明：接种外源微生物可促进堆肥进程,加快有机物分解和堆肥腐熟,消解抗生素等有害物质等作用。餐厨废弃物油脂、盐、水含量高和易酸化等问题会影响好氧发酵过程中微生物的活动。通过特定样品的富集驯化、选择性培养,并结合先进的分子生物学技术,筛选具有不同降解功能的菌株,并以此为基础构建的微生物菌剂能够一定程度上克服餐厨废弃物用于好氧堆肥的限制性问题。主要阐述了餐厨废弃物好氧发酵过程中微生物菌群的演替规律,构建微生物菌剂的菌种类型及功能和不同菌剂对好氧发酵的作用及潜在影响机制,以期能够为相关微生物强化技术的研发提供一些参考。     

餐厨垃圾高温好氧生物减量菌种的筛选及特性

【背景】随着餐厨垃圾产生量的逐步提高,如何实现其快速降解,成为餐厨垃圾处理亟待解决的问题。餐厨垃圾的高温好氧生物减量技术是一种可以快速降解餐厨垃圾的有效方法。【目的】筛选能够适应餐厨垃圾环境且具有高效降解餐厨垃圾中有机物能力的菌株,以提高餐厨垃圾的降解效率和减量效果。【方法】采用温度梯度耐受性实验和餐厨垃圾浸出液高油高盐耐受性实验进行菌种初筛,并利用产酶培养基复筛及餐厨垃圾生物减量实验验证。【结果】通过初筛、复筛和功能验证,最终获得4株生物减量效果优良的菌株N3-1、C7、N3-3和G6-1,其对餐厨垃圾挥发性固体(volatile solid,VS)的降解率分别为36.95%、33.23%、32.83%和31.91%,是对照组的3.02、2.71、2.68和2.61倍。经鉴定,这4株菌分别属于热嗜油地芽孢杆菌(Geobacillus thermoleovorans)、史氏芽孢杆菌(Bacillus smithii)、热解木糖地芽孢杆菌(Geobacillus caldoxylosilyticus)和立陶宛地芽孢杆菌(Geobacillus lituanicus)。【结论】筛选出的4株菌均具有较强的餐厨垃圾原料适应性和高效的生物降解能力,为开发餐厨垃圾高温好氧复合菌剂奠定了基础,并为实现餐厨垃圾减量化、无害化处理和资源化利用提供了技术支持。     

The selection and properties of Penicillium verruculosum mutants with enhanced production of cellulases and xylanases

The paper describes three Penicillium verruculosum 28K mutants with about threefold enhanced production of five industrially important carbohydrases. The two-stage fermentation process that we developed provided a further two- to threefold increase in the production of carbohydrases. Physiological and biochemical studies showed that the synthesis of all five carbohydrases is inducible. Carboxymethylcellulase, xylanase, and -glucanase are synthesized under a common regulatory control, as is evident from the concurrent increase in the synthesis of these enzymes in the presence of microcrystalline cellulose. The synthesis of avicelase and -glucosidase is evidently induced by other cellulose- and hemicellulose-containing compounds present in the fermentation medium and, hence, is regulated independently of the three aforementioned enzymes.__________Translated from Mikrobiologiya, Vol. 74, No. 2, 2005, pp. 172–178.Original Russian Text Copyright © 2005 by Soloveva, Okunev, Velkov, Koshelev, Bubnova, Kondrateva, Skomarovskii, Sinitsyn.     

生物酶改善溶解浆性能的研究进展

溶解浆由高纯度纤维素组成,用于制造再生纤维素纤维、纤维素酯、纤维素醚等材料。溶解浆性能的好坏对后续产品的生产和加工性能有着很大的影响,其中α-纤维素含量、半纤维素含量、浆粕粘度、灰分、过渡金属离子含量、纤维的物理形态、纤维素分子量分布以及溶解浆的反应性能是其重要的性能指标。生物酶因其绿色、温和、高效的特点,在改善溶解浆性能方面有很好的应用前景,并进行了大量的研究。文中介绍了溶解浆的主要性能指标和改善溶解浆性能的生物酶,重点介绍了纤维素酶和木聚糖酶在改善溶解浆性能方面的应用和研究进展,指出了生物酶改善溶解浆性能目前存在的主要问题,提出了该领域的主要研究方向,并对生物酶处理改善溶解浆性能的技术进行了展望。     

Effects of cellulase and xylanase enzymes on the deconstruction of solids from pretreatment of poplar by leading technologies

Comparative data is presented on glucose and xylose release for enzymatic hydrolysis of solids produced by pretreatment of poplar wood by ammonia fiber expansion (AFEX), ammonia recycled percolation (ARP), controlled pH, dilute acid, flowthrough (FT), lime, and sulfur dioxide (SO₂) technologies. Sugar solubilization was measured for times of up to 72 h using cellulase supplemented with β‐glucosidase at an activity ratio of 1:2, respectively, at combined protein mass loadings of 5.8–116 mg/g of glucan in poplar wood prior to pretreatment. In addition, the enzyme cocktail was augmented with up to 11.0 g of xylanase protein per gram of cellulase protein at combined cellulase and β‐glucosidase mass loadings of 14.5 and 29.0 mg protein (about 7.5 and 15 FPU, respectively)/g of original potential glucose to evaluate cellulase–xylanase interactions. All pretreated poplar solids required high protein loadings to realize good sugar yields via enzymatic hydrolysis, and performance tended to be better for low pH pretreatments by dilute sulfuric acid and sulfur dioxide, possibly due to higher xylose removal. Glucose release increased nearly linearly with residual xylose removal by enzymes for all pretreatments, xylanase leverage on glucan removal decreased at high cellulase loadings. Washing the solids improved digestion for all pretreatments and was particularly beneficial for controlled pH pretreatment. Furthermore, incubation of pretreated solids with BSA, Tween 20, or PEG6000 prior to adding enzymes enhanced yields, but the effectiveness of these additives varied with the type of pretreatment. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

产耐碱性β-1，4-聚糖酶芽孢杆菌A-30液体发酵研究

研究了搅拌转速、ＰＨ控制以及插瓶发酵过程中不同时间硫酸铵的补加对β－１,４聚糖酶形成的影响,优化得到Ａ－３０的β０１,４－聚糖酶分批发酵操作条件和初步优化了（ＮＨ４）２ＳＯ４流加发酵条件。研究结果显示麸皮表面有大量Ａ－３０菌体细胞的吸附,搅拌转速对菌体吸附和β－１,４－聚糖酶的形成有明显影响;发酵过程中ＰＨ下降有利于β－１,４－聚糖酶形成。采用了实期以５ｍｌ／ｈ的速率恒速流加,后期测定（ＮＨ４）２     

The effect of anthracene on total dry matter and extracellular enzyme analysis of a starch-amended composting process

Composting technology has been widely used as a cost-effective, eco-friendly, and efficient solid waste management option for the bioremediation of organic pollutants. Most studies have focused on the use of compost in the bioremediation of organic pollutants, without any attention being paid to the effect of the organic pollutants on the composting process. Therefore, in this study, the effect of an organic pollutant (anthracene) on a starch-amended composting process is reported. In order to determine the effect, the total dry matter and the extracellular enzyme analysis (EEA) of starch-specific (α-glucosidase) and -nonspecific (β-glucosidase) substrates were monitored. Over the course of the experiments, there appeared to be some form of anthracene effect on the progression in the total dry matter and the extracellular enzyme analysis (EEA) of the α-glucosidase, particularly in the starch and anthracene–amended composts. The interactions between the starch and the anthracene consistently increased the total dry matter and the α-glucosidase activity of the starch and anthracene–amended composts. This observation is novel, and the kind of interaction is yet unknown. Therefore, detailed investigation to understand the magnitude of such interaction is necessary to potentially resolve these issues.     

Cellulolytic Microbes in the Yanbaru,a Subtropical Rainforest with an Endemic Biota on Okinawa Island,Japan

Cellulolytic microbes in the soil of the Yanbaru, a subtropical forest with an endemic biota, on Okinawa Island, were isolated and characterized in a search for novel microbial strains with biotechnological potential. Soil samples of the Yanbaru were suspended in sterilized water, inoculated on mineral salt agar overlaid with a filter paper as carbon source, and cultivated aerobically at 30 °C. After 2 weeks of cultivation, emerging colonies were isolated and subjected to phylogenetic and enzyme analyses. The phylogenetic analyses revealed bacterial and fungal isolates belonging to nine and three genera respectively. All isolates possessed cellulase activity, and several strains showed strong activity comparable to Trichoderma cellulase. Many isolates also exhibited xylanase activity.     

The Purification of Soybean 11S Globulin with ConA-Sepharose 4B and Sepharose 6B

Kinetics of the acyl transfer catalyzed by Xanthomonas α-amino acid ester hydrolase was studied. The enzyme hydrolyzed d-α-phenylglycine methyl ester (d-PG-OMe) to give equimolar amounts of d-α-phenylglycine and methanol. With d-PG-OMe as an acyl donor and 7-amino-3-deacetoxy-cephalosporanic acid (7-ADCA) as an acyl acceptor, the enzyme transferred the acyl group from d-PG-OMe to 7-ADCA in competition with water. The addition of amine nucleophiles (7-ADCA and 6-aminopenicillanic acid) decreased the molecular activity (k_o) of the enzyme-catalyzed hydrolysis of d-PG-OMe, whereas it did not alter the Michaelis constant (K_M), and plots of l/k_o against the initial concentration of a nucleophile (n_o) gave a straight line. These results support the assumptions that the overall process for hydrolysis and acyl transfer proceeds through a common acyl-enzyme intermediate, that the acylation step of the enzyme is rate-limiting, and that the transfer competes with the hydrolysis of the acyl donor.     

斜卧青霉纤维素酶和木聚糖酶高产菌株的选育

以纤维素酶高产菌株斜卧青霉A50为出发菌株,通过紫外诱变原生质体获得1株木聚糖酶活力提高80%而纤维素酶活力没有改变的6号菌。蛋白质电泳和酶谱检测结果显示,纤维素酶谱基本无差别,而木聚糖酶谱显示6号菌比A50多了一条带。6号菌优化后的产酶培养基组成为:麸皮7%、葡萄糖0.1%,该条件下,纤维素酶活为19.7IU/mL,木聚糖酶活力为215.4IU/mL。     

Characterization of a gene encoding cellulase from uncultured soil bacteria

To detect cellulases encoded by uncultured microorganisms, we constructed metagenomic libraries from Korean soil DNAs. Screenings of the libraries revealed a clone pCM2 that uses carboxymethyl cellulose (CMC) as a sole carbon source. Further analysis of the insert showed two consecutive ORFs (celM2 and xynM2) encoding proteins of 226 and 662 amino acids, respectively. A multiple sequence analysis with the deduced amino acid sequences of celM2 showed 36% sequence identity with cellulase from the Synechococcus sp., while xynM2 had 59% identity to endo-1,4-beta-xylanase A from Cellulomonas pachnodae. The highest enzymatic CMC hydrolysis was observable at pH 4.0 and 45 degrees C with recombinant CelM2 protein. Although the enzyme CelM2 additionally hydrolyzed avicel and xylan, no substrate hydrolysis was observed on oligosaccharides such as cellobiose, pNP-beta-cellobioside, pNP-beta-glucoside, and pNP-beta-xyloside. These results showed that CelM2 is a novel endo-type cellulase.