碱液湿磨预处理稻草的酶解性能

预处理是提高酶法木质纤维素糖化效率的重要加工过程.本论文对稻草进行碱液湿磨预处理和酶解,探讨了预处理碱液浓度对稻草的成分、结构和酶解的影响,同时利用米氏方程对稻草酶解反应进行动力学分析,求出了米氏常数Km 和最大反应速率rmax.实验结果表明,碱液湿磨预处理明显改善了稻草的酶解性能.未处理稻草酶解的还原糖收率为13.4%、Km 为66.8 mg/mL、rmax 为312.5μmol/(min · mL).采用1%NaOH 溶液对稻草进行预处理1h 后,还原糖收率提高到41.4%,Km 减小到15.9 mg/mL, rmax 提高到666.7μmol/(min · mL).预处理过程中木质素去除、纤维素晶体结构消除、底物可及度增加是酶解中还原糖收率和反应速率上升的主要原因.     

城市生物废弃物干式厌氧消化温度实验研究

研究了温度对干式厌氧发酵过程的影响,结果表明温度对pH值影响,高温(55℃)处理pH值迅速达到最低值,中温(35℃)和室温处理相对滞后,室温处理在消化后期出现酸积累现象,pH值维持在较低水平;发酵后期中温处理和高温处理pH值回升.温度对氧化还原电位影响更为明显,氧化还原电位在消化初期下降很快,到达最低点后波动很小.在整个消化过程中的氧化还原电位依次为室温>中温>高温.高温处理VS去除率最高,其值为36.44%;从产气量及甲烷气体含量比较可知,高温处理效率也是最高,其值分别为2548mL、62%,产气时间最长.     

利用餐厨垃圾循环半连续厌氧发酵产氢研究

利用餐厨垃圾采用半连续厌氧发酵进行产氢的研究。实验结果表明以高温(100℃)预处理15 min的厌氧活性污泥为种泥,在温度37℃,pH 6.0左右,较宽的稀释率(1.0~4.0 d-1)范围内,均能较好的实现厌氧发酵产氢。在稀释率D=2.4 d-1下,流出液中乙醇、乙酸、丙酸、丁酸和戊酸的质量分数分别为5.6%、29.6%、5.4%、58.5%和0.9%,产氢过程属于典型的丁酸型发酵,最终氢气的体积分数可达60%,氢气的产生速率为5.49 m3/(m3.d)。将厌氧发酵液相产物作为稀释液返回到反应器中,反应器的产氢能力大幅度的提高,当回流比R=0.8时,最大产氢速率可达10.9 m3/(m3.d),最终氢气的含量可达65%,厌氧发酵反应器的产氢能力提高了约130%。     

基于GET技术的混合厌氧发酵产甲烷研究

GET是一种国外新研发的在田间利用秸秆进行甲烷生产的技术。为了推动该技术在我国的实际应用,本研究以稻草与牛粪为发酵原料,水稻田土壤作为基底,采用正交试验法,测试了温度、发酵基质配比（稻草∶牛粪）和秸秆腐熟剂浓度对混合厌氧发酵产气量的影响。结果表明：各处理因子的产气动态呈“S”型增长模式,但在快速增长时间和增长幅度上有一定区别。30 ℃和40 ℃处理的累积产气量显著高于10 ℃和20 ℃处理的累积产气量;发酵基质配比以1∶1处理的累积产气量最大,分别比1∶0（对照）、1∶2和1∶3配比高34.12%、20.39%和21.78%;秸秆腐熟剂浓度的产气效果在实验范围内随着浓度的增加而增加。厌氧发酵过程中各实验因子下挥发性脂肪酸（VFAs）含量和pH在20 d左右分别有一个高峰和低谷值且都有较大差异,各因子主要是通过在这个时期对厌氧发酵进程的影响而对整个产气量产生影响。综合分析显示稻草厌氧发酵产生物甲烷的最优组合为：温度30 ℃、发酵基质配比1∶1、秸秆腐熟剂浓度8%。     

不同预处理条件对海水养殖废弃物发酵产氢的影响

【目的】利用海水养殖场有机废弃物厌氧发酵产氢,可在减少有机污染物的同时获取氢气。【方法】以海水养殖场有机废弃物为底物,比较嗜热酶(S-TE)、酸、碱、灭菌、微波不同预处理方法对厌氧发酵产氢效果的影响,并对发酵过程中底物性质变化[SCOD、可溶性蛋白质、可溶性糖、pH、VFAs(挥发性脂肪酸)和乙醇]进行探讨。【结果】灭菌预处理产氢效果最好,产氢率为22.0 mL/g VSS,酸处理的效果最差,产氢率为7.6 mL/g VSS。可溶性糖大量消耗之后,氢气不再产生。接种S-TE预处理污泥的底物能更多地释放营养物质,并在整个发酵过程中保持较为稳定的pH值。发酵过程中产生的VFAs主要成分是乙酸,在发酵后期出现乙醇。【结论】灭菌预处理是海水养殖场有机废弃物厌氧发酵产氢的最佳预处理方法,可溶性糖为这一过程主要的营养来源。     

MSW干式厌氧发酵过程中颗粒度影响研究

不同颗粒度干式厌氧发酵处理生物废弃物,对发酵过程有着重要影响,试验表明：不同颗粒度对pH值变化影响较大,较小颗粒度发酵初期下降较快,较大颗粒度pH值变化不显著,而ORP差异比较明显,颗粒度越小,ORP下降越快;蛋白质和还原糖在匀浆状态下初期含量很高,而中后期无太大差异;纤维素酶在高破碎的情况下具有很强的活性,中后期减弱,而果胶酶活力在处理1中初期上升较快,随后变化不显著;VS和CODcr的去除率为处理2效果最好;从产气总量和产气速率比较,处理2为最佳,所产甲炕含量也为最高。     

预处理对泔脚发酵产氢的强化研究

采用热(80℃,15 min)预处理的城市生活垃圾厌氧消化污泥为接种物,考察了不同预处理方法对泔脚中温(36℃)批式发酵产氢的影响。Gompertz模型拟合结果表明:微波850 W,4 min与pH9.0下预处理泔脚的发酵产氢延迟时间λ、最大比产氢率、产氢率、生物气中氢气的最高体积分数分别为:3.47 h,9.43 mL/(g.h),186.23 mL/g及46.0%时,具有更大的产氢优越性。泔脚的发酵产氢过程也是一个酸化过程,发酵产氢结束后,4个预处理方案的发酵产氢余物的pH在4.40~5.00之间,pH均有较大幅度的下降。     

高温油藏内源微生物及其提高采收率潜力研究

大港孔店油田油藏特征、流体和微生物性质分析结果表明,属于高温生态环境,地层水矿化度较低,氮、磷浓度低,而且缺乏电子受体,主要的有机物来源是油气.油田采用经过除油处理的油藏产出水回注方式开发,油层中存在的微生物类型主要是厌氧嗜热菌,包括发酵菌(102个/mL～105个/mL),产甲烷菌(103个/mL);好氧菌主要存在于注水井周围.硫酸盐还原菌(SRB)还原速率0.002 μg S2-/(L·d)～18.9 μg S2-/(L·d),产甲烷菌产甲烷速率0.012 μgCH4/(L·d)～16.2 μgCH4/(L·d).好氧菌能够氧化油形成生物质,部分氧化产物为挥发性脂肪酸和表面活性荆.产甲烷菌在油氧化菌液体培养基中产生CH4,CO2为好氧微生物和厌氧微生物的共同代谢产物.这些产物具有提高原油流动性的作用.用示踪剂研究了注入水渗流方向.通过综合分析,油藏微生物具有较大的潜力,基于激活油层茵的提高采收率方法在该油田是可行的.     

氨水中和Lactobacillus delbrueckii subsp.lactis BME5-18M发酵生成L-乳酸铵的研究

实验确定了Lactobacillus delbrueckiisubsp.lactis BME5-18M接种的最佳种龄为24h.以氨水取代传统的中和剂碳酸钙中和发酵生成的乳酸、调控发酵液的pH,考察了不同pH值对菌体生长和产酸的影响,确定了菌种生长和产酸的较适pH值为6.5.考察了底物流加速度对菌种生长和产酸的影响,对间歇和流加发酵时菌体的生长量和产酸量进行了动力学关联.在较适pH值6.5和较佳流加速度25mL/h条件下,乳酸的产量可达到136.8g/L,产率为1.71 g/(L·h).     

滇池水葫芦固液分离后的沼气发酵研究

在常温下,分离水葫芦为水葫芦渣和水葫芦汁,水葫芦渣与猪粪结合进行批量发酵试验,研究了产气量及原料半纤维素、纤维素、木质素的降解,结果表明原料的产气率为504.04 L/g TS,甲烷含量65%,半纤维素、纤维素降低,木质素升高.对水葫芦汁厌氧发酵的产气及COD降解情况进行了研究,结果表明水葫芦汁的产气潜力为2.192 mL/g,COD降解率达到91.27%.     

乙醇对瘤胃液接种稻秸的体外发酵产物及细菌群落结构的影响

旨在研究乙醇对山羊瘤胃液与水稻秸秆厌氧共培养的影响。利用频繁传代的体外发酵技术和高通量测序方法,分析了短链脂肪酸(SCFA)产量和细菌群落的变化。结果表明,经体外培养传代8次的稻秸发酵液的总短链脂肪酸产量显著高于瘤胃液(P<0.01);与未添加乙醇的稻秸发酵液相比,添加乙醇显著提高了乙酸、戊酸和己酸的比例,降低了丙酸和丁酸的比例(P<0.01),总SCFA产量及异丁酸和异戊酸比例无显著差异。与瘤胃液相比,稻秸发酵液的拟杆菌门(Bacteroidetes)相对丰度下降,厚壁菌门(Firmicutes)相对丰度升高(P<0.05),且添加乙醇显著提高了厚壁菌门和放线菌门(Actinobacteria)的相对丰度(P<0.05);添加乙醇使双歧杆菌属(Bifidobacterium)、未定性的毛螺菌属(unidentified Lachnospiraceae)、产琥珀酸菌属(Succiniclasticum)、脱硫弧菌属(Desulfovibrio)和未定性的梭菌属(unidentified Clostridiales)的相对丰度显著升高(P<0.05)。乙醇使稻秸发酵液的显著性差异物种(Biomarker)增加;稻秸发酵液与瘤胃液亲缘关系较近,而添加乙醇显著改变了细菌区系;短链脂肪酸比例在稻秸发酵液细菌群落多样性中具有重要作用。研究表明,体外频繁传代和添加乙醇可以提高稻秸发酵液的乙酸、戊酸和己酸产量,乙醇改变了稻秸发酵液的细菌群落结构。     

添加剂对水稻秸秆实地混合厌氧发酵产甲烷系统的作用

为提高水稻秸秆利用效率,改进水稻秸秆实地混合厌氧发酵产甲烷技术,本研究开展了添加剂在混合厌氧发酵系统的应用研究。试验选取水稻秸秆和猪粪作为发酵原料,通过分别添加不同的浓度的吐温20和腐植酸,测定甲烷的产气量和浓度、秸秆的降解和土壤肥力的变化,以揭示添加剂类型及其浓度对水稻田实地甲烷生产系统的影响。结果表明:添加剂的掺入并未影响产气的动态变化趋势,但显著地促进产气和提高产气浓度,整体效果由高到低为腐植酸吐温20对照,其中经腐植酸处理的产气量和产气浓度对照相比分别提高了50. 73%和24. 55%。添加剂的掺入有利于水稻秸秆纤维素和半纤维素的降解,但对木质素没有显著影响;其中以0. 15 g/L腐植酸和0. 30 g/L吐温20的降解率最高,相较于对照其纤维素降解率均提高了22. 11%,半纤维素降解率分别提高了107. 13%和98. 39%。添加剂的掺入能显著增加土壤肥力,以0. 15 g/L腐植酸和0. 30 g/L吐温20处理水平的效果最优,相较于对照,其土壤有机质分别增加了29. 63%和23. 72%,全氮分别增加了52. 32%和42. 38%,全磷分别增加了83. 33%和57. 14%。     

Kinetics of rice straw methane fermentation

Summary The kinetics of anaerobic fermentation of rice straw to methane were studied. Rice straw was the only carbon source at influent volatile solid concentrations of 18.9 and 37.8 g/l. Semicontinous runs were carried out at 37°C in laboratory scale perfectly mixed reactors. The Contois' kinetic model constants were calculated from the experimental data. Arefrac tory coefficient was measured (R=0.374) to account for the nonbiodegradable portion of the organic matter of rice straw and incorporated into the kinetic equations. The predicted values of effluent substrate concentration, volumetric methane yield, volumetric methane production rate, and biodegradable conversion efficiency fit well with those measured experi mentally.Percent destruction values of feed constituents were measured.     

一株促生秸秆降解菌的分离与鉴定

针对秸秆处理不当影响全世界环境污染的问题，筛选多功能秸秆降解菌，旨在得到高效降解秸秆且具有促生作用的微生物菌种。结合纤维素钠-刚果红(CMC-Na)平板筛选，通过16S rRNA基因分析，进行菌株鉴定，得到一株具有纤维素降解效果的菌株XJ-132,经16S rRNA基因鉴定为枯草芽胞杆菌(Bacillus subtilis)。与单独施用秸秆处理相比，加入菌株XJ-132 60 d后，秸秆降解率提高21.0%,且对水稻生长促进作用显著，地上、下部鲜重分别增加17.8%和9.6%。水稻种子喷施菌株XJ-132发酵液，低浓度发酵液对种子萌发具有一定促进作用。结果表明，菌株XJ-132可能通过产吲哚乙酸(IAA)、产铁载体、产氨等多种有益物质，降解秸秆的同时促进水稻生长。筛选具有促生作用的秸秆降解菌能够更好地加速秸秆降解，具有广泛的开发利用前景。     

Utilization of agro-industrial waste for xylanase production by Aspergillus foetidus MTCC 4898 under solid state fermentation and its application in saccharification

Xylanase production by Aspergillus foetidus MTCC 4898 was carried out under solid state fermentation using wheat bran and anaerobically treated distillery spent wash. Response surface methodology involving Box–Behnken design was employed for optimizing xylanase production. The interactions among various fermentation parameters viz. moisture to substrate ratio, inoculum size, initial pH, effluent concentration and incubation time were investigated and modeled. The predicted xylanase activity under optimized parameters was 8200–8400 U/g and validated xylanase activity was 8450 U/g with very poor cellulase activity. Crude xylanase was used for enzymatic saccharification of agroresidues like wheat straw, rice straw and corncobs. Dilute NaOH and ammonia pretreatments were found to be beneficial for the efficient enzymatic hydrolysis of all the three substrates. Dilute NaOH pretreated wheat straw, rice straw and corncobs yielded 4, 4.2, 4.6 g/l reducing sugars, respectively whereas ammonia treated wheat straw, rice straw and corncobs yielded 4.9, 4.7, 4.6 g/l reducing sugars, respectively. The hydrolyzates were analysed by HPTLC. Xylose was found to be the major end product with traces of glucose in the enzymatic hydrolyzates of all the substrates.     

Ethanol production from rice straw using optimized aqueous-ammonia soaking pretreatment and simultaneous saccharification and fermentation processes

Rice straw was pretreated using aqueous-ammonia solution at moderate temperatures to enable production of the maximum amount of fermentable sugars from enzymatic hydrolysis. The effects of various operating variables including pretreatment temperature, pretreatment time, the concentration of ammonia and the solid-to-liquid ratio on the degree of lignin removal and the enzymatic digestibility were optimized using response surface methodology. The optimal reaction conditions, which resulted in an enzymatic digestibility of 71.1%, were found to be 69 °C, 10 h and an ammonia concentration of 21% (w/w). The effects of different commercial cellulases and the additional effect of a non-cellulolytic enzyme, xylanase, were also evaluated. Additionally, simultaneous saccharification and fermentation was conducted with rice straw to assess the ethanol production yield and productivity.     

利用光合菌发酵对玉米秸杆进行转化的研究*

系统研究了光合菌在氨法处理和非氨处理两种环境下,在厌氧、好氧、以及自然条件下对玉米秸秆的转化。通过发酵液中还原糖与蛋白质浓度的测定结果的比较、判断、优选出一种最适合条件下光合菌对玉米转化的途径。研究表明,在以氨化法处理的玉米秸秆为底物与光合菌的发酵实验中,发酵液中的还原糖和蛋白质的尝试都要比非氨法条件下玉米秸秆为底物与光合菌发酵实验中的发酵液中的不原糖和蛋白质的浓度高。实验结果证明了转化产生的还原糖、蛋白质都是光合菌能利用的营养成分,由此达到利用光合菌转化玉米秸秆的研究目的。     

The optimized CO2-added ammonia explosion pretreatment for bioethanol production from rice straw

A CO₂-added ammonia explosion pretreatment was performed for bioethanol production from rice straw. The pretreatment conditions, such as ammonia concentration, CO₂ loading level, residence time, and temperature were optimized using response surface methodology. The response for optimization was defined as the glucose conversion rate. The optimized pretreatment conditions resulting in maximal glucose yield (93.6 %) were determined as 14.3 % of ammonia concentration, 2.2 MPa of CO₂ loading level, 165.1 °C of temperature, and 69.8 min of residence time. Scanning electron microscopy analysis showed that pretreatment of rice straw strongly increased the surface area and pore size, thus increasing enzymatic accessibility for enzymatic saccharification. Finally, an ethanol yield of 97 % was achieved via simultaneous saccharification and fermentation. Thus, the present study suggests that CO₂-added ammonia pretreatment is an appropriate process for bioethanol production from rice straw.     

The effects of aqueous ammonia-pretreated rice straw as solid substrate on laccase production by solid-state fermentation

Chemical composition and physical structure of solid substrate have significantly impacts on fermentation performance. The aqueous ammonia was used to pretreat rice straw. Furthermore, the feasibility of pretreatment to improve laccase production was also evaluated in terms of the enzymatic digestibility, chemical structure, physical structure, and laccase production. The results showed that aqueous ammonia pretreatment could modify chemical compositions, destroy rigid structure of the lignocellulosic substrate, increase enzymatic digestibility and change water state, which were beneficial to facilitate the fungus growth and nutrition utilization. Pretreatment of lignocellulosic substrate with aqueous ammonia at 80 °C gave the best effect on laccase production, yielding 172.74 U/g laccase at 14 days, which was 3.4 times higher than that of the control. The aqueous ammonia pretreatment could alternate the physicochemical characteristics of lignocellulosic substrate, resulting in the improved laccase production, which was a promising method that might be explored in solid-state fermentation.

     

Interactions between rumen anaerobic fungi and ciliate protozoa in the degradation of rice straw cell walls

Suspensions of mixed rumen protozoa were added to incubations of the anaerobic fungus Neocallimastix patriciarum with rice straw cell walls. The protozoa did not influence the dry matter lost from the straw, or the solubilization of monosaccharides, but they had a marked effect on the fermentation products formed. Studies with ¹⁴C-labelled protozoa suggested that the presence of protozoa reduced the fungal carboxymethylcellulase activity to around half of that found in pure cultures of the fungus.