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。     

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。     

发酵条件对发酵产氢细菌B49产氢的影响

采用间歇发酵实验，研究了葡萄糖浓度、接种量、温度、氮源、不同有机底物对发酵产氢产酸细菌新菌种IM9(AF481148 in EMBL)生物产氢的影响。结果表明，接种量影响IM9的产氢；IM9生长和产氢适宜温度均为35℃；IM9不能利用无机氮源，而有机氮是IM9生长、产氢的适宜氮源；葡萄糖是IM9发酵产氢的最适宜底物，当浓度为10g／L时，IM9的葡萄糖利用率为100％，氢气得率为1．69molH2／mol glucose；此外，IM9可利用小麦、大豆、玉米、土豆及糖蜜废水和啤酒废水产氢，其中利用糖蜜废水、啤酒废水产氢分别为137．9ml H2／g COD和49．9ml H2／g COD。     

产酸发酵细菌产氢机理探讨

生物制氢技术在世界范围内受到了普通重视，对于生物产氢的机理研究也在不断深入，为生物制氢技术的开发打下了坚实的基础。该文在前人研究成果的基础上，对产酸发酵细菌的多种产氢途径和机理进行了全面探讨。分析认为，在产酸相反应器中的产酸发酵细菌，其主要产氢途径是丙酮酸脱羧产氢和辅酶Ⅰ的氧化与还原平衡调节产氢。通过生化反应的热力学分析证明，即使是氢分压高达0．5个标准大气压时，只要生境中pH值小于4．78，NADH＋H^ 转化为氢的过程就可以顺利进行。     

诱变育种选育高效产氢细菌

从自行研制的CSTR反应器中分离出一株产氢发酵细菌Ethanoligenens sp.ZGX4,以其为出发菌株,进行紫外和亚硝酸复合诱变选育,经过连续传代得到一株遗传稳定性很好的高效产氢突变株YR-3。在培养条件分别为36±1℃,初始pH为6.0,葡萄糖浓度为12g/L,其单位体积产氢量(Y_H_2)为3097.5mL/Lculture,产氢能力比对照提高70.5%,最大产氢速率为36.6mmol/g·drycell·h,比对照高出55.1%;发酵液相末端产物是以乙醇和乙酸为主的典型乙醇型发酵代谢类型。高效产氢耐酸突变体YR-3的释氢能力和产氢速率明显高于野生菌株ZGX4,显示了较强的商业应用潜力,也可为以后进一步探讨研究乙醇型细菌的制氢机制及代谢途径提供物质材料。     

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。     

产氢菌的投加方式对强化发酵菌群产氢的影响

通过间歇培养研究了产氢菌Ethanoligenens sp B49的投加方式对生物制氢反应器的混合发酵菌群生物强化作用的影响.结果表明,产氢菌的投加方式对发酵菌群的产氢能力有显著影响.产氢菌发酵液的直接投加使发酵菌群的产氢能力下降,并引起培养液中发酵产物乙醇和乙酸浓度的显著增加.分析认为,产氢菌发酵液对发酵菌群的末端产物抑制和低pH值抑制作用是导致产氢作用受到抑制的主要原因.离心后单独投加产氢菌菌体可提高发酵菌群的产氢能力,起到强化产氢的作用.投加10.8%的产氢菌强化发酵菌群时,培养45h的累计产氢量为155.0 mL.比强化前发酵菌群培养的产氢量提高了21.5%.因此在利用产氢菌生物强化发酵菌群的研究中,应采用离心分离后单独投加产氢菌菌体的方式进行生物强化.     

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。     

替硝唑片对污泥厌氧发酵产氢作用的研究

研究表明，在污泥中添加替硝唑片可使污泥中的有机物质含量明显增加，特别是可溶的有机物质；替硝唑片对提高污泥产氢效果显著，替硝唑片添加量为1．40g时，污泥的产氢效果最佳，其累积产氢量和产氢潜能分别为580．64mL和98．41mL／gVS；厌氧发酵属于典型的乙醇型发酵；发酵代谢过程主要降解的有机物质为糖类物质，总糖降解率高达70．32％，蛋白质降解率只有26.25％。     

丁酸型产氢-产酸发酵细菌pH生态位探讨

本试验结果认为,细菌在3 86,或4 5>PH>5 3生态位理论。分析原因在于,试验过程中的环境因素———C/N比的降低,氮源物质浓度的提高,相应提高了微生物的合成代谢水平,并且使得细菌发酵过程在pH值较低的环境中,向合成代谢水平较高的丁酸型发酵转变。细菌发生丁酸型发酵是在环境内多种环境因子协同作用下进行的。该试验结果拓宽了前期理论研究中得到的丁酸型发酵生态位范围,为今后相关的理论研究及实际生产提供了理论依据。     

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.     

Accelerated startup of biological hydrogen production process by addition of Ethanoligenens harbinense B49 in a biofilm-based column reactor

Two biofilm-based column reactors with walnut shell (WS) as carrier media were applied for fermentative hydrogen production using glucose as substrate by mixed microbial cultures at the temperature of 35 °C. Pure hydrogen producing bacteria Ethanoligenens harbinense B49 was supplemented into the reacting system periodically or continuously to enhance hydrogen production ability at the startup period. The results showed that the bioreactors supplemented with E. harbinense B49 performed better than the reactor without bacteria addition. Continuous addition mode was recommended, since the hydrogen production performance was better and the operation was easily to be accomplished. The optimal addition amount of pure bacteria was also investigated. The optimal bacterial addition amount was found to be 2.5% which led a better hydrogen production rate. In addition, the bioreactor supplemented with pure bacteria continuously presented a high hydrogen production ability as the specific hydrogen production rate (SHPR) maximized at 1.36 L/g-VSS·d; whereas, the bioreactor without bacteria addition obtained a maximum specific hydrogen production rate of 1.10 L/g-VSS·d. The addition of E. harbinense B49 favored the transformation to ethanol type fermentation in the bioreactor. Thereby, the startup period had been accelerated remarkably.     

Bio-hydrogen production by a new isolated strain Rhodopseudomonas sp. WR-17 using main metabolites of three typical dark fermentation type

In this work, a new strain WR-17 was isolated for photo-fermentative hydrogen production and its hydrogen production capacity was investigated by utilizing main liquid byproducts of three dark fermentation types in batch culture. Experimental results indicated that strain WR-17 was identified as genus Rhodopseudomonas and maximum hydrogen yield of 2.42 mol H₂/mol acetate was obtained when the acetate was used as sole carbon source. Strain WR-17 had an excellent ability of using mixed short chain acids of three typical fermentations such as acetate and ethanol, acetate and butyrate, acetate and propionate. Result demonstrated that the metabolites of butyric acid-type fermentation as substrate is fitting to produce hydrogen and maximum cumulative hydrogen volume of 2156 ml/L-medium was obtained when acetate of 30 mmol/L and butyrate of 15 mmol/L were used. Therefore, butyric acid-type fermentation has great potential for further obtaining high hydrogen yield by the combining photo-fermentation.     

Application of rice rhizosphere microflora for hydrogen production from apple pomace

The combination of substrate materials and bacteria is an important factor affecting conversion technology for biological hydrogen production. We performed anaerobic hydrogen fermentation of apple pomace wastes using rhizosphere bacterial microflora of rice as the parent inoculum. In the vial test, the optimal condition for hydrogen fermentation was initial pH 6.0, 35 °C, and 73.4 g pomace per liter of medium (equivalent to 10 g-hexose/L). In the batch experiment (pH 6.0, temperature 35 °C) the hydrogen yield reached 2.3 mol-H₂/mol-hexose. The time course of biogas production and PCR-DGGE analysis suggest that Clostridium spp. decomposed degradable carbohydrates rapidly and a part of the refractory carbohydrate (e.g. pectin) gradually in the apple pomace slurry. In addition to hydrogen, volatile fatty acids (VFAs) were produced in the anaerobic fermentation of apple pomace, which can be a substrate for methane fermentation. The rice rhizosphere can be a promising source of inoculum bacteria for hydrogen fermentation in combination with plant material waste like apple pomace.     

提高木糖代谢能力的酿酒酵母Y5-X3的初步构建

为获得能够发酵木糖和葡萄糖的新菌种,文中采用实验室专利菌株Saccharomyces cerevisiae Y5为宿主菌,分别采用热带假丝酵母(Candida tropicalis)和休哈塔假丝酵母(Candida shehatae)作为木糖代谢关键酶XYL1,XYL2的来源,利用酵母整合载体将XYL1,XYL2基因导入S.cerevisiae Y5基因组,同时超表达S.cerevisiae Y5内源木酮糖激酶基因XKS1,获得了酿酒酵母新菌种S.cerevisiae Y5-X3。结果表明,S.cere-visiae Y5-X3在摇瓶共发酵3%葡萄糖和2%木糖时能够利用木糖产乙醇,木糖消耗量和乙醇产量分别比宿主菌提高102.93%和12.54%,发酵2%木糖时木糖的消耗量是宿主菌的2.7倍。XKS1基因的超表达促进了木糖代谢,大大减少了木糖醇积累,提高了乙醇产量。     

Selection of natural bacterial communities for the biological production of hydrogen

Current processes used for the production of hydrogen consume a great part of the energy they produce and/or depend on fossil fuel consumption, making them inefficient and harmful to the environment. Obtaining hydrogen from living systems by fermentation of organic matter considered waste is a promising alternative for the future. Especially when you take into account that the biological production of hydrogen is intrinsically linked to the degradation of said organic matter. In this paper, we explore the efficiency of different bacterial communities (also called consortia) for anaerobic fermentation of carbohydrates. The evaluated consortia were obtained from soil, commercial compost and sludge from a sewage treatment plant. The cultures that produced the highest amounts of hydrogen were those in which the inoculums used came from sludge and compost. Both reached a maximum accumulated concentration of approximately 30% of biological hydrogen in the gas mixture on day 8 of the fermentation process, as estimated by gas chromatography.     

Enhancement of fermentative hydrogen/ethanol production from cellulose using mixed anaerobic cultures

Batch tests were conducted to evaluate the enhancement of hydrogen/ethanol (EtOH) productivity using cow dung microflora to ferment α-cellulose and saccharification products (glucose and xylose). Hydrogen/ethanol production was evaluated based on hydrogen/ethanol yields (HY/EY) under 55 °C at various initial pH conditions (5.5–9.0). Our test results indicate that cow dung sludge is a good mixed natural-microflora seed source for producing biohydrogen/ethanol from cellulose and xylose. The heat-pretreatment, commonly used to produce hydrogen more efficiently from hexose, applied to mixed anaerobic cultures did not help cow dung culture convert cellulose and xylose into hydrogen/ethanol. Instead of heat-pretreatment, the mixed culture received enrichments cultivated at 55 °C for 4 days. Positive results were observed: hydrogen/ethanol production from fermenting cellulose and xylose was effectively enhanced at increases of 4.8 (ethanol) to 8 (hydrogen) and 2.4 (ethanol) to 15.6 (hydrogen) folds, respectively. In which, the ethanol concentration produced from xylose reached 4–4.4 g/L, an output comparable to that of using heat-treated sewage sludge and better than that (1.25–3 g/L) using pure cultures. Our test results show that for the enriched cultures the initial cultivation pH can affect hydrogen/ethanol production including HY, EY and liquid fermentation product concentration and distribution. These results were also concurred using a denaturing gradient gel electrophoresis analysis saying that both cultivation pH and substrate can affect the enriched cow dung culture microbial communities. The enriched cow dung culture had an optimal initial cultivation pH range of 7.6–8.0 with peak HY/EY values of 2.8 mmol-H₂/g-cellulose, 5.8 mmol-EtOH/g-cellulose, 0.3 mol-H₂/mol-xylose and 1 mol-EtOH/mol-xylose. However, a pH change of 0.5 units from the optimal values reduced hydrogen/ethanol production efficiency by 20%. Strategies based on the experimental results for optimal hydrogen/ethanol production from cellulose and xylose using cow dung microflora are proposed.     

Hydrogen production from organic waste

The extraction of pure hydrogen from the fermentation of household waste by a mixed anaerobic bacterial flora is demonstrated. Simulated household waste (600 g) was fermented in a bioreactor, which was continuously sparged with nitrogen (30 ml/min) fed in from the bottom. The gas stream from the biorector passes through a sulphide trap (ZnO) and then through a heated palladium–silver membrane reactor to separate hydrogen from the gas stream. In this way, waste remediation and biological hydrogen production is combined in a process where a large proportion of the hydrogen produced can be collected, free of other gaseous species from the fermentation.