C/N对细菌产氢发酵类型及产氢能力的影响

反应基质中的C/N比作为影响因子,参与细菌的产能代谢过程,主要作用于微生物的自身合成代谢过程和有机物在微生物体内的生物氧化过程。乙醇型发酵过程中由于物质和能量转化问高度平衡细胞合成代谢处于较低的水平,而丁酸型发酵过程中,由于NADH H参与细胞合成代谢。所以发酵基质内C／N比过低,过剩的N源物质进一步促进了微生物细胞的合成代谢。并且导致的发酵类型向丁酸型发酵转变的现象,是微生物种群维持“内平衡”的适应性结果。分析认为反应基质中的C/N比作为影响因子,是作用于系统发酵产氢过程稳定性的主要因素之一。在试验及生产过程中,为了得到最佳产氢发酵类型一乙醇型发醇,应严格控制反应系统底物环境内C／N≥200,将微生物细胞合成代谢过程控制在较低的水平,在提高系统产氢能力及其稳定性的同时,降低系统剩余污泥的产生量。     

Fe对产氢发酵细菌发酵途径及产氢能力影响

经过对20株产氢发酵细菌的静态发酵试验，结果发现加入Fe的培养液中细菌发酵由原来的丁酸型发酵过程向乙醇型发酵过程转化；在有机物发酵产氢的两个主要途径中，Fe为必要成分之一，其参与促进酶促反应的进行。在相似培养条件下，单质Fe与Fe^2 均可诱导细菌代谢向乙醇型发酵转化，其中单质Fe的作用能力优于Fe^2 ；在细菌代谢过程中，单质Fe具有提高细菌发酵产氢能力的作用。     

乙醇型发酵与丁酸型发酵产氢机理及能力分析

氢气是一种新型清洁能源，方便快捷的制氢方法正日益受到重视，产氢－产酸发酵过程中氢气产生的主要途径为乙醇型发酵过程和丁酸型发酵过程，在对这两种发酵类型产氢机理及能力的理论研究及对比试验中，发现乙醇型发酵途径发酵产氢能力要优于丁酸型发酵过程（平均为25％－40％），并且该文将pH值和氧化还原电位（ORP）作为综合环境因子考察，通过与发酵过程产氢量指标相配合分析，认为乙醇型发酵过程是一种较佳的有机物生物发酵制氢途径。     

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。     

丁酸型发酵产氢的运行稳定性

着重对发酵法生物制氢反应系统的丁酸型发酵的运行稳定性进行了研究分析。结果表明,在有机负荷大于21kgCOD/m3·d的条件下,丁酸型发酵具有不稳定性,在负荷冲击下容易转变为丙酸含量较高的发酵类型,从而导致系统产氢能力的下降。分析认为,NADH/NAD+的平衡调节能力是影响系统运行稳定性的一个关键因素。在高负荷条件下,由于丁酸型发酵的产丁酸过程不能氧化过剩的NADH+H+,导致产乙酸过程生成的剩余NADH+H+在系统内大量积累,使反应系统难以达到氧化还原的平衡状态,最终影响了系统的稳定运行。     

木糖厌氧发酵产氢特性研究

以木糖作为厌氧发酵产氢底物,热预处理(100℃,处理20 min)的厌氧颗粒污泥作为接种物,研究了中温条件(37℃)下厌氧发酵产氢特性.结果表明,当反应进行至50 h时,累积产氢量最大,为81.11 mL;乙酸、丁酸和乙醇是液相末端产物中的主要物质,其中乙酸和丁酸的浓度分别为1290 mg/L和1225 mg/L,发酵类型是典型的丁酸型发酵;反应体系的pH值开始降低,最后稳定在4.40左右,形成一个稳定的缓冲体系.     

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。     

pH值调控对发酵产氢的影响

利用厌氧活性污泥作产氢接种物，发酵有机质产生氢气，一般是在酸性条件下进行的。以厌氧活性污泥作接种物，有机酸为基质，在厌氧、恒温25℃、不同的pH值下，启动发酵产氢，以及监测产氢过程中的pH值变化，得出pH值过高时，有大量的甲烷生成，pH值过低时，则对产氢细菌不利，难于产氢。启动发酵产氢时，pH值不宜底于4．3，较为适宜的产氢pH值范围4．5～5．5。     

光合细菌小分子酸醇产氢试验研究

以光合产氢混合菌群为研究对象,研究了光合细菌在乙酸、乙醇、乳酸、丁酸几种小分子脂肪酸条件下菌体的生长和产氢特性,详细考察了乙酸和丁酸对光合产氢细菌生长和产氢的影响.研究发现,乙酸、丁酸既是光合细菌良好的生长碳源,也是高效氢供体,光合细菌在乙酸和丁酸条件下产氢率分别达到2.05和2.81molH2/mol.光合细菌以乙酸和丁酸产氢时,乙酸和丁酸的最佳添加浓度均为40mmol/L;光合细菌在乳酸条件下有较高的生长活性,但乳酸并不是光合细菌高效氢供体,光合细菌在乳酸条件下产氢活性较低;乙醇既不是光合细菌良好生长碳源,也不是高效氢供体,乙醇对光合细菌的生长和产氢均有较强的抑制作用.     

维生素B4对秸秆类生物质光发酵产氢的影响

以玉米秸秆类生物质为产氢原料,研究维生素B4对HAU-M1光合细菌生长和产氢过程的影响规律。结果表明,当维生素B4浓度为75 mg/L时,光合细菌生长情况最好,细菌干重最大值为0.934 g/L;维生素B4浓度为100 mg/L时,氢气累积产量达178 mL,比对照组显著提高了43.8%,对光合细菌产氢的促进效果最好;添加维生素B4对HAU-M1光合细菌发酵产氢过程的pH值影响显著,可减弱发酵液酸化,有利于光合细菌发酵产氢。显见,维生素B4对HAU-M1光合细菌生长及秸秆类生物质光合产氢具有明显的促进作用,可为进一步研究开发秸秆类生物质光合细菌发酵产氢工艺技术提供科学参考。     

Hydrogenase activity monitoring in the fermentative hydrogen production using heat pretreated sludge: A useful approach to evaluate bacterial communities performance

The conversion of organic compounds into H₂ has received increasing attention. Enrichment of inocula by heat pretreatment eliminates non-spore forming hydrogen consuming microorganisms and promotes spore germination in genus Clostridium, which is known as one of the key hydrogen producers. Useful information about metabolic pathway is provided by some intermediate metabolites, such as: acetic, propionic, butyric and formic acids. The increase of acetic/butyric acids ratio indicates H₂ production in heat pretreated inoculum when compared to untreated inoculum in the same cultivation conditions. The effect of heat pretreatment on inocula and consequently on the performance of bacterial communities responsible for H₂ production was monitored through the measurement of the level of hydrogenase gene expression, as well as through the content and distribution of volatile fatty acids. The acetic acid type fermentation was followed by the microorganisms presented in untreated and heat pretreated sludge. The medium containing untreated sludge presented a ratio of acetic/butyric acid of approximately 4, the same parameter was 7 when heat pretreated sludge was employed. The level of hydrogenase gene expression tripled when heat pretreated inoculum was used, indicating a higher production of H₂.     

有机负荷对杂交狼尾草和牛粪混合发酵系统内微生物群落的影响

选取杂交狼尾草和牛粪为原料,研究有机负荷和原料配比对混合原料发酵系统内微生物群落组成的影响.研究结果表明,属水平上系统内优势细菌有梭菌属(Clostridum)、普雷沃菌属(Prevotella)、互养棍状菌属(Syntrophorhabdus)、假单胞菌属(Pseudomonas)和噬蛋白质菌属(Proteiniph...     

Capnophilic lactic fermentation and hydrogen synthesis by Thermotoga neapolitana: An unexpected deviation from the dark fermentation model

The heterotrophic bacterium Thermotoga neapolitana produces hydrogen by fermentation of organic substrates. The process is referred to as dark fermentation and is typically complemented by production of acetic acid. Here we show that synthesis of products derived by reductive metabolism of pyruvate, mainly lactic acid, occurs to the detriment of acetic acid fermentation when the cultures of the thermophilic bacterium are flushed by saturating level of CO₂. Sodium bicarbonate in a very narrow range of concentrations (∼14 mM) also causes the same metabolic shift. The capnophilic (CO₂-requiring) re-orientation of the fermentative process toward lactic acid does not affect hydrogen productivity thus challenging the currently accepted dark fermentation model that predicts reduction of this gas when glucose is converted into organic products different from acetate.     

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。     

Enzymatic dynamics of microbial acid tolerance response (ATR) during the enhanced biohydrogen production process via anaerobic digestion

Organic acid especially butyric acid accumulation can inhibit fermentative hydrogen production. However acid tolerance could be increased when mixed cultures adapted to butyric acid stress in an intentional and continuous operation. As compared to the original cultures, the yield and productivity of hydrogen were enhanced by 56.5% with more acid production and a higher HAc/HBu (acetic acid/butyric acid) of 1.33 by the evolved cultures. The enhancement was due to the increase of acid tolerance with more acid tolerance response (ATR) induction. Later the representative enzymatic ATR systems were investigated in this study. It was found that two ATR systems including H⁺-ATPase and hydrogenase played important roles in hydrogen evolution and the evolved cultures had a higher overall induction. Furthermore, the dynamics of the ATR systems indicated that hydrogen-producing microorganisms with acid tolerance could prepare themselves better against self-produced acid stress and the evolved cultures had a better stress anticipation possibly due to the bacterial communities change via adaptive evolution.     

Inhibitory effect of ethanol,acetic acid,propionic acid and butyric acid on fermentative hydrogen production

The inhibitory effect of added ethanol, acetic acid, propionic acid and butyric acid on fermentative hydrogen production by mixed cultures was investigated in batch tests using glucose as substrate. The experimental results showed that, at 35 °C and initial pH 7.0, during the fermentative hydrogen production, the substrate degradation efficiency, hydrogen production potential, hydrogen yield and hydrogen production rate all trended to decrease with increasing added ethanol, acetic acid, propionic acid and butyric acid concentration from 0 to 300 mmol/L. The inhibitory effect of added ethanol on fermentative hydrogen production was smaller than those of added acetic acid, propionic acid and butyric acid. The modified Han–Levenspiel model could describe the inhibitory effects of added ethanol, acetic acid, propionic acid and butyric acid on fermentative hydrogen production rate in this study successfully. The modified Logistic model could describe the progress of cumulative hydrogen production.     

A sequential approach to uncapping of theoretical hydrogen production in a sulfate-reducing bacteria-based bio-electrochemical system

The presence of undesired methanogens with Sulfate-reducing bacteria (SRBs) is a serious challenge faced by the bioelectrochemical system (BES). In the present study, we investigate the impact of ammonia pre-treated electrodes on hydrogen production in a 600 ml anaerobic (BES) enriched with sulfate-reducing bacteria (SRBs) to inhibit the CH₄ production for achieving the theoretical H₂ production. The highest hydrogen production of 3.67 ± 0.31 M/M of glucose was recorded in the BES. The BES completely inhibited the growth of methanogens after the 7^th cycle of operation. The higher hydrogen production efficiency of BES can be justified by assuming a higher hydrogen mass transfer from the electrode surface to the biofilm. In presence of sulfate, acetate acid type of the fermentation was dominating in hydrogen production, while limitation of SO₄^2? switch over to the dominance of butyric acid type fermentative hydrogen production. Despite the sign of change in the acetate to butyric acid type metabolism, the BES system was able to uncap the theoretical hydrogen production. The notable change in vector orientation of H₂, butyric acid, and hexanoic acid inferring the significant differences in the microbial community adapted on the electrodes in the R–NH₃ and R-Cont. SEM image clearly showing ammonia-treated electrode harbour more microbial growth on the electrode surface. The ratio obtained for CH₃ and CH₂ for the R–NH₃ and R-CONT of 1.316 and 1.755 respectively by FTIR stretching vibrations showing the difference in the bacterial species adapted on the bioanodes. Cumulative hydrogen production data was computed to confirm its validity of the Gompertz model, Richard model, and Logistic model. The Richard model was found in the best-fitted models for cumulative hydrogen production.     

Influence of butyrate on fermentative hydrogen production and microbial community analysis

The effect of butyrate on hydrogen production and the potential mechanism were investigated by adding butyric acid into dark fermentative hydrogen production system at different concentrations at pH range of 5.5–7.0. The results showed that under all the tested pH from 5.5 to 7.0, the addition of butyric acid can inhibit the hydrogen production, and the inhibitory degree (from 10.5% to 100%) increased with the increase of butyric acid concentration and with the decrease of pH values, which suggested that the inhibition effect is highly associated with the concentration of undissociated acids. Substrate utilization rate and VFAs accumulation also decreased with the addition of butyric acid. The microbial community analysis revealed that butyrate addition can decrease the dominant position of hydrogen-producing microorganisms, such as Clostridium, and increase the proportion of other non-hydrogen-producing bacteria, including Pseudomonas, Klebsiella, Acinetobacter, and Bacillus.     

Enhanced hydrogen and volatile fatty acid production from sweet sorghum stalks by two-steps dark fermentation with dilute acid treatment in between

This study investigated the potential of hydrogen and volatile fatty acid coproduction from two steps dark fermentation with dilute acid treatments of the residual slurry after 1st step fermentation. Sweet sorghum stalks (SS) was used as substrate along with Clostridium thermosaccharolyticum as production microbe. Residual lignocelluloses after 1st step fermentation were treated for 1 h by sulfuric acid concentration of 0.25, 0.5, 1.0, 1.5, 2.0 and 2.5% (w/v) with different reaction temperature of 120, 90 and 60 °C were studied. The optimum severity conditions for the highest yield of products found from the treatment acid concentration of 1.5% (w/v) at 120 °C for 10 g/L of substrate concentration. Experimental data showed that two-step fermentation increased 76% hydrogen, 84% acetic acid and 113% of butyric acid production from single step. Maximum yields of hydrogen, acetic acid and butyric acid were 5.77 mmol/g-substrate, 2.17 g/L and 2.07 g/L respectively. This two-step fermentation for hydrogen and VFA production using the whole slurry would be a promising approach to SS biorefinery.