Determination of the cellulase activity distribution in Clostridium thermocellum and Caldicellulosiruptor obsidiansis cultures using a fluorescent substrate

This study took advantage of resorufin cellobioside as a fluorescent substrate to determine the distribution of cellulase activity in cellulosic biomass fermentation systems. Cellulolytic biofilms were found to express nearly four orders greater cellulase activity compared to planktonic cultures of Clostridium thermocellum and Caldicellulosiruptor obsidiansis, which can be primarily attributed to the high cell concentration and surface attachment. The formation of biofilms results in cellulases being secreted close to their substrates, which appears to be an energetically favorable stategy for insoluble substrate utilization. For the same reason, cellulases should be closely associated with the surfaces of suspended cell in soluble substrate-fed culture, which has been verified with cellobiose-fed cultures of C. thermocellum and C. obsidiansis. This study addressed the importance of cellulase activity distribution in cellulosic biomass fermentation, and provided theoretical foundation for the leading role of biofilm in cellulose degradation. System optimization and reactor designs that promote biofilm formation in cellulosic biomass hydrolysis may promise an improved cellulosic biofuel process.     

高效木薯渣分解复合菌群RXS的构建及其发酵特性研究

从富含腐烂纤维质的环境中取样,通过以木薯渣及滤纸为碳源的蛋白胨纤维素培养基不断地富集培养,构建了一组高效稳定的纤维质分解复合菌群.考察了该复合菌群对不同纤维质底物的分解性能及其在木薯渣水解过程中主要参数的变化.研究发现该复合菌群对滤纸、脱脂棉、微晶纤维素、麦秸秆和木薯渣等原料均能够进行有效的降解.在该复合菌群应用于木薯渣的水解过程中,监测发现纤维素酶、半纤维素酶、果胶酶等关键酶的酶活力分别在第2～3 d达到最大值34.4、90.5和15.8U;经过10 d的发酵后,木薯渣中的纤维素、半纤维素及木质素分别降解了79.8%、85.9%和19.4%,且木薯渣的失重高达61.5%;此外,代谢产物主要是乙酸、丁酸、己酸和甘油;而溶解性COD、总糖和总挥发酸的变化表明第2 d时木薯渣的水解率最高.上述结果表明,该复合菌群能够有效地水解木薯燃料酒精生产过程中的废弃物木薯渣,并有望用于木薯渣高效沼气发酵的前处理中.     

Tween 80和鼠李糖脂对稻草酶解的影响

采用纤维素酶促水解的方法,以稻草为底物,探讨了添加化学表面活性剂Tween 80和生物表面活性剂鼠李糖脂对酶解过程的糖产率、酶稳定性、纤维素转化率的作用以及对酶动力学特征和酶在纤维素上吸附的影响.结果表明,不同浓度的Tween 80和鼠李糖脂对稻草酶解有不同程度的促进,添加0.016%和0.048%Tween 80使糖产率分别提高18.07%和11.98%,而添加0.01%和0.03%鼠李糖脂使糖产率分别增加了23.01%和22.16%,相比较鼠李糖脂的效果更好.表面活性剂能有效增强酶的稳定性,添加高浓度表面活性剂的酶稳定性优于添加低浓度表面活性剂,添加浓度为0.048%的Tween 80得到最高相对CMCA(羧甲基纤维素酶活)108.06%和最高相对FPA(滤纸酶活)80.26%.表面活性剂能提高酶解反应的纤维素转化率,而且添加鼠李糖脂的转化率明显高于Tween 80.表面活性剂不仅能够提高最大反应速度并使米氏常数变大,而且显著地降低了纤维素酶在纤维素上的吸附.     

Combination effect of pH and acetate on enzymatic cellulose hydrolysis

The productivity and efficiency of cellulase are significant in cellulose hydrolysis. With the accumulation of volatile fatty acids (VFAs), the pH value in anaerobic digestion system is reduced. Therefore, this study will find out how the pH and the amount of acetate influence the enzymatic hydrolysis of cellulose. The effects of pH and acetate on cellulase produced from Bacillus coagulans were studied at various pH 5-8, and acetate concentrations (0-60 mmol/L). A batch kinetic model for enzymatic cellulose hydrolysis was constructed from experimental data and performed. The base hypothesis was as follows: the rates of enzymatic cellulose hydrolysis rely on pH and acetate concentration. The results showed that the suitable pH range for cellulase production and cellulose hydrolysis (represents efficiency of cellulase) was 2.6-7.5, and 5.3-8.3, respectively. Moreover, acetate in the culture medium had an effect on cellulase production (K1= 49.50 mmol/L, n=1.7) less than cellulose hydrolysis (K1=37.85 mmol/L, n=2.0). The results indicated that both the pH of suspension and acidogenic products influence the enzymatic hydrolysis of cellulose in an anaerobic environment. To enhance the cellulose hydrolysis rate, the accumulated acetate concentration should be lower than 25 mmol/L, and pH should be maintained at 7.     

Combination e ect of pH and acetate on enzymatic cellulose hydrolysis

The productivity and e ciency of cellulase are significant in cellulose hydrolysis. With the accumulation of volatile fatty acids (VFAs), the pH value in anaerobic digestion system is reduced. Therefore, this study will find out how the pH and the amount of acetate influence the enzymatic hydrolysis of cellulose. The e ects of pH and acetate on cellulase produced from Bacillus coagulans were studied at various pH 5–8, and acetate concentrations (0–60 mmol/L). A batch kinetic model for enzymatic cellulose hydrolysis was constructed from experimental data and performed. The base hypothesis was as follows: the rates of enzymatic cellulose hydrolysis rely on pH and acetate concentration. The results showed that the suitable pH range for cellulase production and cellulose hydrolysis (represents e ciency of cellulase) was 2.6–7.5, and 5.3–8.3, respectively. Moreover, acetate in the culture medium had an e ect on cellulase production (KI = 49.50 mmol/L, n = 1.7) less than cellulose hydrolysis (KI = 37.85 mmol/L, n = 2.0). The results indicated that both the pH of suspension and acidogenic products influence the enzymatic hydrolysis of cellulose in an anaerobic environment. To enhance the cellulose hydrolysis rate, the accumulated acetate concentration should be lower than 25 mmol/L, and pH should be maintained at 7.     

低温下玉米秸秆与牛粪混合厌氧发酵产气特性研究

通过静态试验,探究在15℃的低温条件下,经过预处理的玉米秸秆和牛粪,在不同混合配比下厌氧发酵的产气特性。试验结果表明:将牛粪与秸秆按1∶1配比的产气效果最好,相对于配比为2∶1、3∶1的累积产气量分别高出52.66%和73.60%。随着发酵过程进行,COD波动很大。发酵过程中碳源主要由玉米秸秆提供,秸秆所占比例越大,COD降解率越高。日产气量达到高峰的时间稍滞后于VFA,秸秆比例越大的组别,VFA整体处于越高水平。而辅酶F420与产气量之间关系不显著。发酵初期CMC酶活力变化显著,且以玉米秸杆与牛粪配比为1∶1为配比的一组CMC酶活力高于其余2组,最高值达到了15.042 U/g。     

秸秆发酵产氢的影响因素研究

以加热预处理的厌氧活性污泥为接种物,以秸秆（稻草、麦杆）为发酵底物进行微生物产氢实验,研究了不同秸秆粒径、纤维素酶用量、预处理方法、发酵液pH值对秸秆发酵产氢速率以及产氢气量的影响.其中颗粒粒径为170 μm的稻草经过NaOH预处理,按照底物与纤维素酶用量1∶1的比例酶解,进而发酵产氢的单位总产氢量为90.5 mL/g,最大单位产氢速率为0.58 mL/(h·g).     

The climate change challenge and transitions for radical changes in the European steel industry

This paper presents ideas pertaining to transitions that are envisaged in the steel industry from Cleaner Production (CP) to Systems Innovation. Limits of the socio-technical system and the climate change challenge will induce changes in the production, distribution and consumption patterns of steel and other materials. Insights from industrial economics and evolutionary theory on innovation for sustainable development are needed to assess the rationale behind the adoption and diffusion of new breakthrough technologies named Ultra Low CO₂ Steel making (ULCOS). Evolution in material consumption patterns deserves a special research agenda which focuses upon the long term evolution of the consuming sectors as major changes in the infrastructure and products that support our many energy dependent services (mobility, shelter, heat, light, etc.) are expected. These changes will be significantly amplified by greenhouse gas emission constraints.     

鼠李糖脂逆胶束体系中纤维素酶的后萃研究

采用生物表面活性剂鼠李糖脂(rhamnolipid,RL)构建RL/异辛烷/正己醇的逆胶束体系,并研究了该体系中纤维素酶后萃过程的影响因素.分别考察了后萃水相pH值、振荡时间、离子种类和强度以及添加短链醇对纤维素酶的后萃率和酶活回收率的影响.结果表明,后萃水相最佳pH值为7.0,振荡时间以30 min为最佳,后萃水相中离子强度以0.15 mol·L-1KCl最佳,正丁醇的最佳添加量为2%.在最佳实验条件下,纤维素酶的后萃率和酶活回收率分别可以达到76.22%和93.39%.生物表面活性剂RL构建的逆胶束体系对纤维素酶的后萃效果较佳,且RL具有高生物降解性,低临界胶束浓度等优点,应用前景广阔.     

利用柑桔加工过程中的废物生产纤维素酶发酵饲料

利用柑桔加工过程中废物提取有用物质后 ,再进行固体发酵生产纤维酶发酵饲料的研究 ,筛选到一株具有较高纤维素酶的青霉菌株。其最适固体培养基配方为 :桔皮∶豆饼∶麸皮 =15∶1∶1,碳酸铵质量分数为 3% ,物料∶水 =2∶1,pH值为 4 5 ;最佳产酶条件为 :培养温度为 30℃ ,培养时间为 84h左右。为柑桔加工过程中所产的废物提供了一条变废为宝 ,解决环境污染的新途径。      

Soil enzyme activities and their indication for fertility of urban forest soil

To reveal the biological characteristics of urban forest soil and the effects of soil enzyme on soil fertility as well as the correlation between physicochemical properties and enzyme activities, 44 urban forest soil profiles in Nanjing were investigated. Basic soil physicochemical properties and enzyme activities were analyzed in the laboratory. Hydrogen peroxidase, dehydrogenase, alkaline phosphatase, and cellulase were determined by potassium permanganate titration, TTC (C₁₉H₁₅N₄·Cl) colorimetry, phenyl phosphate dinatrium colorimetry, and anthrone colorimetry, respectively. The result showed that soil pH, organic carbon (C), and total nitrogen (N) had great effects on hydrogen peroxidase, dehydrogenase, and alkaline phosphatase activities in 0–20 cm thick soil. However, pH only had great effect on hydrogen peroxidase, dehydrogenase, and alkaline phosphatase activities in 20–40 cm thick soil. Hydrogen peroxidase, dehydrogenase, and alkaline phosphatase were important biological indicators for the fertility of urban forest soil. Both in 0–20 cmand 20–40 cmsoil, soil enzyme system (hydrogen peroxidase, dehydrogenase, alkaline phosphatase, and cellulase) had a close relationship with a combination of physicochemical indicators (pH, organic C, total N, available K, available P, cation exchange capacity (CEC), and microbial biomass carbon (C_mic)). The more soil enzyme activities there were, the higher the fertility of urban forest soil. __________ Translated from Urban Environment & Urban Ecology, 2007, 20(4): 4–6, 9 [译自: 城市环境与城市生态]     

Long-term stabilization of enzyme activity by immobilizing enzyme colony in polymaleimidostyrene-modified micelles

Amperometric biosensors fabricated by immobilizing enzyme colony in polymaleimidostyrene-modified micelles exhibited good sensitivity and great long-term stability. To our knowledge, there has not been a general design for various enzyme biosensors. Enzyme micelle membrane which is different from any of other conventional immobilization methods, is an innovative way and will be a well-developed biosensor technology to provide rapid and reliable measurements. The potential of using enzyme micelle membrane to fabricate biosensors will be of great hope.     

Attributional life cycle assessment of woodchips for bioethanol production

Besides the apparent need to reduce greenhouse gas emissions, other important factors contributing to the renewed interest in biofuels are energy security concerns and the need of sustainable transportation fuel. Nearly 30% of the annual CO₂ emissions in the U.S. come from the transportation sector and more than half of the fuel is imported. Biofuels appear to be a promising option to reduce carbon dioxide emissions, and the reliance on imported oil concomitantly. The interest on (ligno) cellulosic ethanol is gaining momentum as corn-based ethanol is criticized for using agricultural outputs for fuel production. Among many lignocellulosic feedstocks, woodchips is viewed as one of the most promising feedstocks for producing liquid transportation fuels. The renewable and carbon neutral nature of the feedstocks, similar chemical and physical properties to gasoline, and the low infrastructure cost due to the availability of fuel flex vehicles and transportation networks make (ligno) cellulosic bioethanol an attractive option. An in-depth LCA of woodchips shows that harvesting and woodchips processing stage and transportation to the facility stage emit large amount of environmental pollutants compared to other life cycle stages of ethanol production. Our analysis also found that fossil fuel consumption and respiratory inorganic effects are the two most critical environmental impact categories in woodchips production. We have used Eco-indicator 99 based cradle-to-gate LCA method with a functional unit of 4 m³ of dry hardwood chips production.     

高效稳定纤维素分解混合菌群的筛选及分解特性研究

以高纤维素素含量的土壤为筛选源,利用定向筛选技术,经过多代淘汰,最终筛选出一组木质纤维素分解混合菌群P-C。混合菌群的生长曲线没有明显的分界线,在培养4至5天时,出现了短暂的平稳期,在培养第7天时出现峰值,此时混合菌群的纤维素酶活最高。以天然纤维素为产酶碳源时,混合菌群纤维素酶活明显高于以人工纤维素为产酶碳源时的酶活,以秸秆为碳源时的纤维素酶活是以滤纸为碳源时的1.5倍。混合菌群纤维素酶的最适pH是6,最适酶促反应温度是45℃,但在35℃-40℃时也具有较高酶活。pH=6,糖化48 h时,发酵液糖浓度最高。     

壬基酚降解酶的固定化及特性研究

用海藻酸钠包埋法对壬基酚(NP)降解酶进行了固定化研究,并对固定化酶的最适反应温度和最适pH、及其稳定性进行了探讨。实验结果表明,用3%海藻酸钠、2%CaCl2溶液固定化的酶活力最高;固定化NP降解酶的最适反应温度为35℃,最适pH为7.0,其热稳定性和pH稳定性都有一定程度的提高。。     

含水率对生活垃圾甲烷化过程的影响

采用批式实验,通过分析产气量和气、液相组成的变化,比较了含水率分别在35%、65%～70%（田间持水率）、 80%和>95%（饱和含水率）的条件下,不同食物类废物和纤维素类废物含量的典型法国生活垃圾和中国生活垃圾以及纤维素类废物厌氧产甲烷过程的差异.结果表明,对于含易腐有机物的生活垃圾,提高含水率能够削弱VFA等中间产物对水解酸化和甲烷化的抑制作用：典型法国生活垃圾大量产甲烷所需的含水率不应低于80%,而有机物含量更高的典型中国生活垃圾直至含水率达到96%时,反应体系内快速产甲烷过程才能得以进行;同时,提高生活垃圾的含水率也有利于加速其厌氧产甲烷过程和提高甲烷的最终产量,含水率>95%的法国生活垃圾反应体系的最终甲烷产量是含水率为80%时产量的1.6倍.而对于纤维素类废物,提高含水率则能改善水分在水解和酸化过程中的可获得性,从而增大甲烷化的底物可获得量,含水率>95%的纤维素类废物反应体系的甲烷最终产量是含水率为65%时产量的3.8倍.     

糖蜜、淀粉与乳品废水厌氧发酵法生物制氢

采用连续流厌氧发酵法研究了糖蜜废水、淀粉废水与牛奶废水生物制氢,讨论了有机物中3大类物质的厌氧发酵产氢的可行性,并对几种底物的产氢稳定性进行了探讨.结果表明,糖蜜废水与淀粉废水都是较好的厌氧发酵法生物产氢底物,在3大类有机物中碳水化合物是目前技术条件下最具有可能性的原材料.在碳水化合物中,溶解性好的糖比溶解性差的淀粉在目前的技术条件下具有生物产氢可行性,而淀粉比溶解性糖更具有产氢前景.不同底物厌氧生物制氢的生态位范围有所不同,对于溶解性好的糖,稳定运行的工程控制参数为pH 4.5±0.3,而溶解性较差的淀粉废水为pH 4.0±0.2;厌氧发酵产氢的ORP值也随底物的不同而不同,但总体必须低于-220mV,在-300mV左右时较好.牛奶废水不适用于作为CSTR反应器中发酵法生物制氢底物.     

微生物燃料电池的研究综述

微生物燃料电池是一种利用微生物的催化作用,将燃料中的化学能转化为电能,同时又可以处理废水的新型技术,具有显著的环境效益和经济效益。本文对微生物燃料电池的基本原理进行了详细的叙述,对一些影响微生物燃料电池在处理污水时发电的基本因素做了较全面的比较,同时也探讨了一些现阶段微生物燃料电池的瓶颈问题。展望了微生物燃料电池(MFCs)这一绿色技术的良好的发展前景。     

中国粮食生产区域集中化的演化趋势

农业生产的区域专业化与地域分工是农业现代化的一个重要标志,也是未来中国农业发展的必由之路。受农业生产市场化程度以及耕地保护、农业补贴、价格保护等政策因素的影响,中国目前区域专业化的方向和地域分工取向尚不清晰,不过农产品生产的集中化趋势却为回答这一问题提供了重要线索。论文以稻谷、小麦和玉米3种主要粮食作物为研究对象,利用省域单元1981—2013年的产量数据,运用集中化指数方法揭示1980年以来中国粮食生产区域集中化程度的演变特征和演化趋势。结果表明：中国粮食生产的集中化程度明显上升,3种粮食作物中小麦生产的集中化程度最高,并呈相对稳定的集中化趋势,而稻谷和玉米生产的集中化程度则经历了一个“U”型变化过程,意味着稻谷和玉米生产在这一阶段发生了空间重组;依据集中化趋势,稻谷生产向黑龙江、湖南、江西等地集中,小麦生产向河南、山东、安徽等传统小麦产区集中,玉米生产则向黑龙江、内蒙古等地集中,预示未来极有可能在这些地区及其周边形成专业化粮食产区。加强这些地区的农田基本建设将有助于推进中国粮食生产的地区专业化与地域分工。