厌氧高效产氢细菌的筛选及其耐酸性研究

采用厌氧Hungate技术 ,从生物制氢反应器厌氧活性污泥中分离到 18株发酵产氢细菌 ,并从中优选出 1株高效产氢细菌B4 9。通过间歇试验 ,B4 9获得最大比产氢速率QH2 为 2 5 .0mmol/g·h ,单位体积产氢量YH2 为 1813.8mL/L ,氢气含量为 6 4 .15 %。B4 9菌株为乙醇型发酵产氢细菌 ,具有良好的耐酸性 ,在 pH3.3仍能生长。发酵产氢和细菌生长的最适 pH值约为 3.9～ 4 .2。     

秸秆发酵产氢的碱性预处理方法研究

以麦秆、稻草和滤纸为发酵底料,以厌氧活性污泥为接种物,采用不同的预处理方法去除木质素并提高纤维素的降解率,从而提高其发酵产氢能力。试验表明对于相同的底料,经过NaOH预处理和纤维素酶解后的还原糖含量、总产气量、总产氢量和氢气浓度都要高于经过氨水预处理的底料,而未经过预处理的底料发酵产氢能力最差。利用10g经过NaOH预处理的麦秆和稻草,经纤维素酶解后在发酵产氢过程中的降解率分别为23.2%和12.5%,总产氢量分别为363.3mL和254.9mL,发酵产气中氢气浓度分别为23.8%和29.1%。发酵液相中主要产物为乙醇、乙酸和丁酸。     

厨余和污泥不同混合比例碱处理产氢特性研究

以厨余垃圾和污泥为反应底物,加热预处理的污泥为发酵接种物,考察了碱处理下厨余与污泥不同混合比例的发酵产氢特性。结果表明:不同pH碱液对厨余垃圾进行预处理后,其效果以pH=13时最佳,预处理3h后SCOD和还原糖含量分别为31316.8mg/L和5.54mg/mL;碱预处理后的污泥与厨余联合发酵能够改善物料的营养平衡,缩短反应延迟时间到1h内;当厨余与污泥混和比例为5:1时为本试验最佳的试验条件,其氢气含量、比产氢速率峰值和氢产率分别为52.69%,1.73mL H_2/(h·gVS)和50.27mL H_2/gVS。     

预处理温度对活性污泥发酵产氢特性的影响

为寻求适宜的种泥热处理方法,利用摇瓶发酵实验,考察了城市污水处理厂好氧活性污泥分别经65、80、95、110℃热处理30min后,其利用葡萄糖发酵产氢的特性。结果表明:在初始pH=7.0、葡萄糖浓度10g/L、接种量2gMLVSS/L条件下,35℃培养72h,经65℃和95℃处理的种泥表现出较好的发酵产氢性能,其葡萄糖的氢气转化率分别达到1.08和1.11mol/mol,污泥的比产氢率分别为8.36和9.05mmol/gMLVSS;经65℃预处理的种泥发酵体系,表现为丁酸型发酵,其葡萄糖降解率和最大产氢速率分别高达82%和11.29mL/h,而经95℃预处理的种泥发酵体系则呈现混合酸发酵特征,其葡萄糖转化率和最大产氢速率分别仅为76%和4.45mL/h。     

Fe3O4纳米颗粒对玉米秸秆光发酵产氢的影响

以HAU-M1光合菌群作为发酵细菌,以玉米秸秆为发酵底物,研究Fe₃O₄纳米颗粒对光发酵产氢过程的影响。结果表明:粒径60 nm的Fe₃O₄纳米颗粒浓度为100 mg/L时,比产氢量达到(46.68±1.00)mL/g VSS,与对照组的(35.07±0.56)mL/g VSS相比提升(33.11±0.01)%,此时的能量转化率也提高33.10%。产氢动力学分析结果也表明Fe₃O₄纳米颗粒对反应体系有明显的影响,粒径60 nm的Fe₃O₄纳米颗粒浓度为100 mg/L时,最大产氢潜能和最大产氢速率分别为46.97 mL/g VSS和1.06 mL/(g VSS·h)。适宜的Fe₃O₄纳米颗粒的粒径和浓度能显著促进光发酵产氢能力,而浓度过高则会产生抑制作用。     

丁酸型产氢细菌WN9利用谷糠发酵的产氢特征

该研究从牡丹江江滨公园的河道底泥样中筛选获得1株丁酸型发酵产氢细菌的新菌株Clostridium butyricum WN9,并分别以葡萄糖和小米内、外壳谷糠为底物进行发酵产氢实验。实验结果表明:以葡萄糖为底物时,最大比产氢率为1.89 mol/mol;以小米内、外壳谷糠为底物时,内壳谷糠更易被利用,最适宜的内、外壳谷糠浓度分别为50,30 g/L,最大比产氢率分别为20.1,12.7 mL/g;低初始pH值条件(pH6.0)有利于提高谷糠转化效率,当内、外壳谷糠浓度均为50 g/L,初始pH值为6.0时,最大比产氢率分别提高至21.5,15.5 mL/g。     

聚球藻自相发酵产氢研究

研究了培养基的pH值、盐度和氮源等对聚球藻生长及自相发酵产氢的影响。发现聚球藻在弱碱性时(pH=7.5~8)不能正常生长,当pH值高于8.5时藻才能实现富集,当碱性进一步增强到pH值9.5时藻生长状态最佳。收获藻液置于黑暗厌氧条件下利用自身氢酶进行自相发酵产氢,单位干重的产氢量达到22.25mL/g。聚球藻无法适应高盐度环境,在盐度较低情况下(0.154 mmol/L)才能迅速生长,得到自发酵产氢最大值为25.68 mL/g。加入无机氮源能明显提高聚球藻的生长速率及生物质产量,但对随后产氢效果有抑制作用。     

生物制氢反应器产氢产乙酸菌群对挥发酸的转化

采用间歇培养的方式,利用取自生物制氢反应器的厌氧活性污泥考察了活性污泥中产氢产乙酸菌群对乙醇、乙酸、丙酸、丁酸、戊酸和乳酸的转化和产氢。结果表明,培养时间为44h时,厌氧活性污泥发酵葡萄糖的累计产气量为356mL,累计产氢量为209mL,氢气含量为58.7%。发酵产物的组成成分乙醇为427.1mg/L、乙酸为716.5mg/L、丙酸为172.5mg/L、丁酸为689.4mg/L、戊酸为123.6mg/L。发酵生物制氢反应器厌氧活性污泥中产氢产乙酸菌群能够对乙醇和乳酸进行产氢产乙酸转化,厌氧污泥转化乙醇形成的乙酸含量约为270mg/L,累计产氢量为15mL;转化乳酸形成的乙酸含量约为190mg/L,累计产氢量为7mL。厌氧污泥不能对乙酸、丙酸、丁酸和戊酸进行产氢产乙酸转化,培养过程中也没有气体生成,分析认为产氢产乙酸菌群对挥发酸的转化不是发酵生物制氢反应器产氢的主要途径。     

荷叶发酵产沼气的研究

以荷叶为原料对其发酵产沼气进行了研究。以稻草秸秆为对照,活性污泥为接种物,对荷叶厌氧发酵产沼气的能力进行了评估,并研究了荷叶对稻草秸秆发酵产沼气的影响。荷叶在37℃条件下发酵35 d,沼气产气量为167.0 mL/g,沼气中甲烷含量为65.72%,分别高出稻草秸秆15.57%,12.25%。荷叶与稻草秸秆按1∶1进行混合发酵时沼气产气率为184.5 mL/g,沼气中甲烷含量为70.68%,分别高出稻草秸秆27.68%,20.72%。荷叶发酵前后TS,VS含量变化分别为3.77%,15.17%,发酵前后pH维持在7左右。结果表明,荷叶在沼气生产上有很大潜力。     

肉座菌Z28产纤维素酶工艺研究

采用液体发酵法,对肉座菌(Hypocrea sp.)Z28的最适产纤维素酶条件进行了研究.结果表明:在以稻草粉为碳源,硫酸铵为氮源,接种量2%,发酵时间6d,温度为30℃,pH值6.0,转速为180r/min的条件下,纤维素酶活力最高,其CMCase活力为66.35U/mL,FPA活力为13.75U/mL,β-葡萄糖苷酶活力为14.26U/mL.     

Microwave-assisted alkali pretreatment of rice straw to promote enzymatic hydrolysis and hydrogen production in dark- and photo-fermentation

Rice straw was pretreated by microwave-assisted alkali to improve saccharification in enzymatic hydrolysis and hydrogen yield in combined dark- and photo-fermentation in this paper. A maximum reducing sugar yield of 69.3 g/100 g TVS was obtained when 50 g/l rice straw was pretreated by microwave heating for 15 min at 140 °C in 0.5% NaOH solution and then enzymatically hydrolyzed for 96 h. When hydrolyzed rice straw was used for hydrogen production by anaerobic bacteria in dark-fermentation, a maximum hydrogen yield of 155 ml/g TVS was obtained. The residual solution (mainly acetate and butyrate) from dark-fermentation was reutilized for hydrogen production by immobilized photosynthetic bacteria in photo-fermentation. By combination of dark- and photo-fermentation, the maximum hydrogen yield was greatly enhanced to 463 ml/g TVS, which is 43.2% of the theoretical hydrogen yield.     

Effects of operation conditions on enzymatic hydrolysis of high-solid rice straw

In the present work, the performances of high-solids enzymatic hydrolysis of rice straw were experimentally studied. Moreover, the heat flow and weight loss of the pre- and post-hydrolysis of rice straw were measured by the simultaneous thermogravimetric analyzer. It is found that the output and yield of reducing sugar initially increased and then reduced with an increase of supplied water, hydrolysis temperature and substrate mesh, while the cellulose content in residual substrate had an opposite trends. Furthermore, a high enzyme loading resulted in a constant saccharification efficiency of cellulose. The output and yield of the sugar, 77.7 mg/(g substrate) and 0.44 g/g, were respectively achieved under the optimal conditions of the supplied water of 1.0 ml/(g substrate), enzymolysis temperature of 50 °C, rice straw of 60 mesh and the cellulase dosage of 2.0 mg/(g substrate). Simultaneous thermogravimetric analysis on the rice straw indicates that the chemical compositions of rice straw changed after enzymolysis.     

Thermophilic hydrogen fermentation from Korean rice straw by Thermotoga neapolitana

Rice straw, a low-cost lignocellulosic biomass was used as feedstock for thermophilic hydrogen fermentation by Thermotoga neapolitana. Hydrogen production, the growth and cellulose digestibility of the hyperthermophile in batch mode from untreated as well as chemically pretreated (ammonia and dilute sulfuric acid) Korean rice straws were investigated. Pretreatment method using combination of 10% ammonia and 1.0% dilute sulfuric acid was developed to increase the digestibility of rice straw for the hyperthermophilic H₂ fermentation and to decrease the time consumption. In a typical fermentation using raw rice straw, 29% of the substrate was digested and 2.3 mmol H₂/g straw of hydrogen yield was consistently obtained. Compared with the pretreatments using only ammonia or dilute sulfuric acid, the combined pretreatment method using both chemical agents significantly increases the digestibility of rice straw with 85.4% of substrate consumption. H₂ production on rice straw from this combined pretreatment showed the highest yield (2.7 mmol H₂/g straw) and the highest sugar conversions (72.9% of glucose and 95.7% of xylose).     

Biological hydrogen production from steam-exploded straw by simultaneous saccharification and fermentation

Hydrogen was produced by simultaneous saccharification and fermentation from steam-exploded corn straw (SECS) using Clostridium butyricum AS1.209. Effect of various process parameters, such as solid to liquid ratio, enzyme loading and initial pH, etc., were examined with respect to maximum hydrogen productivity which was obtained by fitting the cumulative hydrogen production data to a modified Gompertz equation. Maximum specific hydrogen production rate and maximal hydrogen yield were 126 ml/g VSS d and 68 ml/g SECS, respectively. The yield of soluble metabolites was 197.7 mg/g SECS. Acetic acid accounted for 46% of the total was the most abundant product and this shows that hydrogen production from SECS was essentially acetate-type fermentation. Hydrogen production by simultaneous saccharification and fermentation of SECS has the predominance of short lag-stage and high maximum specific hydrogen production rate and it was a promising method for hydrogen production and straw biomass conversion.     

Hydrogen production by anaerobic co-digestion of rice straw and sewage sludge

In this study, the rich carbon content of rice straw and the high nitrogen content of sewage sludge make the straw a good potential substrate and the sludge a viable inoculum for biohydrogen production. Two treatment conditions for the sewage sludge (raw and heat-treated) were used in the present experiments. Batch test using a mixture of rice straw and sewage sludge were carried out to investigate the optimum carbon to nitrogen (C/N) ratio for effective biohydrogen production. The experimental results indicate that untreated sludge could be used as the inoculum for efficient hydrogen production when mixed with the appropriate proportion of rice straw. According to our results, biogas and hydrogen production in all raw sludge cases ramped up more quickly and also exhibited longer and higher hydrogen production in comparison with heat-treated cases. At the C/N ratio of 25 in untreated sludge, hydrogen production was 33% higher than heat-treated one. Additionally, under the same conditions, high and stable hydrogen content (58%) and the maximal hydrogen yield (0.74 mmol H₂/g-VS added straw) were obtained.     

碱性双氧水预处理玉米秸秆性质研究

使用碱性双氧水对玉米秸秆进行预处理,可以有效提高秸秆的酶解效果。实验表明,最优预处理参数为使用分别占秸秆质量16%的 H₂O₂和25.6% 的NaOH,于40℃下预处理秸秆24 h。对经不同预处理剂处理后的秸秆进行酶解,发现NaOH及碱性双氧水预处理秸秆的酶解还原糖产量为7.48 g/L和8.26 g/L,而经H₂O及H₂O₂预处理秸秆的还原糖产量仅为1.35 g/L和1.59 g/L。通过木质纤维素含量及SEM分析发现,氢氧化钠主要作用为溶解秸秆中的木质素及半纤维素,而双氧水的存在则会破坏秸秆表面结构。计算秸秆预处理前后质量损失发现,双氧水的存在不能显著提高秸秆预处理后的质量损失,但会氧化分解被氢氧化钠溶解的大分子物质,对此过程机理及产物还需进行深入的研究。     

猪粪发酵产氢潜力的研究

采用批量发酵工艺，以解猪粪为原料，进行了厌氧发酵产氢的研究，发酵料液pH值控制在5．0左右，实验结果表明，鲜猪粪的产氢潜力为127ml/g(TS)和158ml/g(VS).     

Thermophilic dark fermentation of untreated rice straw using mixed cultures for hydrogen production

Biohydrogen production from untreated rice straw using different heat-treated sludge, initial cultivation pH, substrate concentration and particle size was evaluated at 55 °C. The peak hydrogen production yield of 24.8 mL/g TS was obtained with rice straw concentration 90 g TS/L, particle size <0.297 mm and heat-treated sludge S1 at pH 6.5 and 55 °C in batch test. Hydrogen production using sludge S1 resulted from acetate-type fermentation and was pH dependent. The maximum hydrogen production (P), production rate (R_m) and lag (λ) were 733 mL, 18 mL/h and 45 h respectively. Repeated-batch operation showed decreasing trend in hydrogen production probably due to overloading of substrate and its non-utilization. PCR-DGGE showed both hydrolytic and fermentative bacteria (Clostridium pasteurianum, Clostridium stercorarium and Thermoanaerobacterium saccharolyticum) in the repeated-batch reactor, which perhaps in association led to the microbial hydrolysis and fermentation of raw rice straw avoiding the pretreatment step.     

Simultaneous biohydrogen (H2) and bioplastic (poly-β-hydroxybutyrate-PHB) productions under dark,photo, and subsequent dark and photo fermentation utilizing various wastes

The present study is focused on bio hydrogen (H₂) and bioplastic (i.e., poly-β-hydroxybutyrate; PHB) productions utilizing various wastes under dark fermentation, photo fermentation and subsequent dark-photo fermentation. Potential bio H₂ and PHB producing microbes were enriched and isolated. The effects of substrate (rice husk hydrolysate, rice straw hydrolysate, dairy industry wastewater, and rice mill wastewater) concentration (10–100%) and pH (5.5–8.0) were examined in the batch mode under the dark and photo fermentation conditions. Using 100% rice straw hydrolysate at pH 7, the maximum bio H₂ (1.53 ± 0.04 mol H₂/mol glucose) and PHB (9.8 ± 0.14 g/L) were produced under dark fermentation condition by Bacillus cereus. In the subsequent dark-photo fermentation, the highest amounts of bio H₂ and PHB were recorded utilizing 100% rice straw hydrolysate (1.82 ± 0.01 mol H₂/mol glucose and 19.15 ± 0.25 g/L PHB) at a pH of 7.0 using Bacillus cereus (KR809374) and Rhodopseudomonas rutila. The subsequent dark-photo fermentative bio H₂ and PHB productions obtained using renewable biomass (i.e., rice husk hydrolysate and rice straw hydrolysate) can be considered with respect to the sustainable management of global energy sources and environmental issues.