光合菌生物制氢技术

简要分析了光合细菌产氢的主要影响因素,介绍了国内外光合细菌生物制氢技术的研究和应用现状,并对光合制氢技术的发展趋势和应用前景进行了评述。     

Photoproduction of hydrogen by photosynthetic bacteria from sewage and waste water

Numerous prokaryotes, belonging to physiologically and taxonomically different groups, are able to produce hydrogen. Some photosynthetic bacteria have the property of light-dependent production of hydrogen from organic substrates. We isolated several photosynthetic purple and green bacteria from enrichment cultures made from the water of a waste-water pond of a cool-drink refilling station. After testing them for their ability to use various organic compounds as carbon source, and sulphide, thiosulphate and organic compounds as electron donor, we selected the fastest-growing isolate, aRhodopseudomonas, for a study of its ability to produce molecular hydrogen in presence of light. Immobilized cells of this isolate produced significant amounts of hydrogen from both sewage and waste water     

Photoproduction of molecular hydrogen from waste water of a sugar refinery by photosynthetic bacteria

Summary From enrichment cultures inoculated with water and sediments of a waste-water pond of a sugar refinery several photosynthetic nonsulphur bacteria have been isolated and tested for the ability to produce molecular hydrogen in the light. Strains have been found that utilize the freshly used, untreated waste substrate with higher yields than the laboratory strain used so far. Under the test conditions one strain showed higher hydrogen production rates from waste water than from any synthetic substrate.     

Acetate as a carbon source for hydrogen production by photosynthetic bacteria

Hydrogen is a clean energy alternative to fossil fuels. Photosynthetic bacteria produce hydrogen from organic compounds by an anaerobic light-dependent electron transfer process. In the present study hydrogen production by three photosynthetic bacterial strains (Rhodopseudomonas sp., Rhodopseudomonas palustris and a non-identified strain), from four different short-chain organic acids (lactate, malate, acetate and butyrate) was investigated. The effect of light intensity on hydrogen production was also studied by supplying two different light intensities, using acetate as the electron donor. Hydrogen production rates and light efficiencies were compared. Rhodopseudomonas sp. produced the highest volume of H2. This strain reached a maximum H2 production rate of 25 ml H2 l(-1) h(-1), under a light intensity of 680 micromol photons m(-2) s(-1), and a maximum light efficiency of 6.2% under a light intensity of 43 micromol photons m(-2) s(-1). Furthermore, a decrease in acetate concentration from 22 to 11 mM resulted in a decrease in the hydrogen evolved from 214 to 27 ml H2 per vessel.     

Photobiological hydrogen production: Recent advances and state of the art

Photobiological hydrogen production has advanced significantly in recent years, and on the way to becoming a mature technology. A variety of photosynthetic and non-photosynthetic microorganisms, including unicellular green algae, cyanobacteria, anoxygenic photosynthetic bacteria, obligate anaerobic, and nitrogen-fixing bacteria are endowed with genes and proteins for H₂-production. Enzymes, mechanisms, and the underlying biochemistry may vary among these systems; however, they are all promising catalysts in hydrogen production. Integration of hydrogen production among these organisms and enzymatic systems is a recent concept and a rather interesting development in the field, as it may minimize feedstock utilization and lower the associated costs, while improving yields of hydrogen production. Photobioreactor development and genetic manipulation of the hydrogen-producing microorganisms is also outlined in this review, as these contribute to improvement in the yield of the respective processes.     

暗发酵-光发酵两阶段联合生物制氢技术研究进展

随着能源紧缺的日益加剧,以及化石燃料燃烧引起的环境问题逐渐突显,氢能作为一种清洁可再生能源越来越受到青睐。生物制氢与热化学及电化学制氢相比其反应条件温和、低耗、绿色,是一项非常有应用前景的技术。生物制氢从广义上可以分为暗发酵和光发酵产氢两种,其中暗发酵微生物可以利用有机废弃物产生氢气以及有机酸等副产物,光合细菌在光照和固氮酶的作用下可以将暗发酵产生的有机酸继续用于产氢,因此两种发酵产氢方式相结合可以提高有机废物的资源化效率。将近年来暗发酵-光发酵两阶段生物制氢技术进行整理分析,从其产氢机理、主要影响因素、暗发酵-光发酵产氢结合方式(两步法、混合培养产氢)几个方面进行阐述,最后指出该技术面临的挑战。     

Engineering photosynthetic light capture: impacts on improved solar energy to biomass conversion

The main function of the photosynthetic process is to capture solar energy and to store it in the form of chemical 'fuels'. Increasingly, the photosynthetic machinery is being used for the production of biofuels such as bio-ethanol, biodiesel and bio-H₂. Fuel production efficiency is directly dependent on the solar photon capture and conversion efficiency of the system. Green algae (e.g. Chlamydomonas reinhardtii ) have evolved genetic strategies to assemble large light-harvesting antenna complexes (LHC) to maximize light capture under low-light conditions, with the downside that under high solar irradiance, most of the absorbed photons are wasted as fluorescence and heat to protect against photodamage. This limits the production process efficiency of mass culture. We applied RNAi technology to down-regulate the entire LHC gene family simultaneously to reduce energy losses by fluorescence and heat. The mutant Stm3LR3 had significantly reduced levels of LHCI and LHCII mRNAs and proteins while chlorophyll and pigment synthesis was functional. The grana were markedly less tightly stacked, consistent with the role of LHCII. Stm3LR3 also exhibited reduced levels of fluorescence, a higher photosynthetic quantum yield and a reduced sensitivity to photoinhibition, resulting in an increased efficiency of cell cultivation under elevated light conditions. Collectively, these properties offer three advantages in terms of algal bioreactor efficiency under natural high-light levels: (i) reduced fluorescence and LHC-dependent heat losses and thus increased photosynthetic efficiencies under high-light conditions; (ii) improved light penetration properties; and (iii) potentially reduced risk of oxidative photodamage of PSII.     

Characteristics of anaerobic ammonia removal by a mixed culture of hydrogen producing photosynthetic bacteria

It is known that the presence of ammonia inhibits hydrogen production by photosynthetic bacteria. In order to avoid it, a two-step process containing ammonia removal and hydrogen production was investigated in this study. Firstly, the effects of carbonate presence on ammonia removal by photosynthetic bacteria were investigated by the vial tests because it is known that the uptake of volatile fatty acids (VFAs) sometimes requires carbonate. The results of them showed that the presence of carbonate promoted the uptake of VFAs and ammonia. Especially, the uptake of propionate and/or butyrate required the presence of carbonate. The results of the batch experiments of two-step hydrogen production showed that the depletion of ammonia triggered hydrogen evolution. Herein, the presence of albumin did not inhibit hydrogen evolution and preferably it increased the hydrogen production rate. And the VFA-C/NH4-N ratio in substrate fed into two-step hydrogen production process should be more than 6.0.     

正交实验优选光合细菌混合菌群产氢的最佳条件

对光合细菌混合菌群产氢影响因子进行了实验研究。通过单因素实验和正交实验, 系统考察了碳源、氮源、碳源浓度、氮源浓度、初始pH值、光照方式、接种量等因素对产氢量的影响, 实验得出最佳工艺条件为: 采用3号菌群, 碳源为葡萄糖, 碳源浓度为3 g/L, 氮源为尿素, 氮源浓度为9 g/L, 接种量为10%, pH值为8.5, 光照方式为12 h光照-12 h黑暗交替光照, 培养温度为30°C。菌种、碳源、碳源浓度、氮源是影响产氢量的重要因素。     

Progress toward a biomimetic leaf: 4,000 h of hydrogen production by coating‐stabilized nongrowing photosynthetic Rhodopseudomonas palustris

Intact cells are the most stable form of nature's photosynthetic machinery. Coating‐immobilized microbes have the potential to revolutionize the design of photoabsorbers for conversion of sunlight into fuels. Multi‐layer adhesive polymer coatings could spatially combine photoreactive bacteria and algae (complementary biological irradiance spectra) creating high surface area, thin, flexible structures optimized for light trapping, and production of hydrogen (H₂) from water, lignin, pollutants, or waste organics. We report a model coating system which produced 2.08 ± 0.01 mmol H₂ m^?2 h^?1 for 4,000 h with nongrowing Rhodopseudomonas palustris, a purple nonsulfur photosynthetic bacterium. This adhesive, flexible, nanoporous Rps. palustris latex coating produced 8.24 ± 0.03 mol H₂ m^?2 in an argon atmosphere when supplied with acetate and light. A simple low‐pressure hydrogen production and trapping system was tested using a 100 cm² coating. Rps. palustris CGA009 was combined in a bilayer coating with a carotenoid‐less mutant of Rps. palustris (CrtI^?) deficient in peripheral light harvesting (LH2) function. Cryogenic field emission gun scanning electron microscopy (cryo‐FEG‐SEM) and high‐pressure freezing were used to visualize the microstructure of hydrated coatings. A light interaction and reactivity model was evaluated to predict optimal coating thickness for light absorption using the Kubelka‐Munk theory (KMT) of reflectance and absorptance. A two‐flux model predicted light saturation thickness with good agreement to observed H₂ evolution rate. A combined materials and modeling approach could be used for guiding cellular engineering of light trapping and reactivity to enhance overall photosynthetic efficiency per meter square of sunlight incident on photocatalysts. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Perspectives on cultivation strategies and photobioreactor designs for photo-fermentative hydrogen production

Photosynthetic bacteria have considerable biotechnological potential for biological hydrogen production due to higher substrate conversion efficiency and hydrogen yield. Phototrophic fermentation using photosynthetic bacteria has a major advantage of being able to further convert the byproducts originating from dark fermentation (e.g., volatile fatty acids) to hydrogen. Through the combination of dark and photo-fermentation processes, organic feedstock is fully converted into gaseous product (H₂) at the highest possible H₂ yield, with significant reduction of chemical oxygen demand (COD). The performance of photo-fermentation is highly dependent on the medium composition, culture conditions, and photobioreactor design. Therefore, this article provides a critical review of the effects of key factors affecting the photo-hydrogen production efficiency of photosynthetic bacteria, and also summarizes the strategies being applied in promoting the performance of photo-fermentation.     

Structure of a microbiol mat at Great Sippewissett Marsh, Cape Cod, Massachusetts

The microzination of phototrophic bacteria in a flat laminated microbiol mat at Great Sippewissett Salt Marsh on Cape Cod, Massachusetts, was studied using a combination of scanning and transmission electron microscopy, light microscopy and photosynthetic pigment analysis. Comparison of pigment content and ultrastructural information from electron microscopy of thin sections allowed us to determine the major groups of photosynthetic bacteria present. The approximately 1-cm-thick mat is located in sandy intertidal sediments of the marsh and comprised four to five distinctly colored layers. The uppermost brown layer contained Lyngbya, Nostoc, Phormidium (cyanobacteria) and Navicula (diatom) species. An intermediate bluish-green layer was dominated by Oscillatoria species. A central pink layer contained purple sulfur bacteria such as Amoebobacter, Thiocapsa, Chromatium and Thiocystis species, Below this was a distinctive orange layer, formed largely by one species of purple sulfur bacteria, Thiocapsa pfennigii. The lowermost and thinnest layer contained green sulfur bacteria of the genus Prosthecochloris, a very small prosthecate species with numerous knobby projections; this layer was not always present. Below this, where pigments were generally absent, were dark gray and black iron sulfide-rich sediments. Remnants of older decayed mats could be found deeper in the sediment. Extensive production of microbial extracellular polymers in all layers appeared to be responsible for attachment of cells to sand grains, for lamination of layers and for structural integrity of the mat as a whole. Below the layer of green sulfur bacteria, binding of sediment by microbial polymers ceased abruptly. Possibly in response to decreasing light penetration, the mean size of bacterial cells decreased in successively deeper layers. In the lowest layer where light penetration was very low, green sulfur bacteria with highly convoluted surfaces occurred. The increase in cell surface area-to-volume ratio may allow such organisms to survive at low light levels.     

THE PENETRATION OF LUMINOUS BACTERIA BY THE AMMONIUM SALTS OF THE LOWER FATTY ACIDS : PART I. GENERAL OUTLINE OF THE PROBLEM,AND THE EFFECTS OF STRONG ACIDS AND ALKALIES.

It is shown that disappearance of the light of luminous bacteria may be used as a criterion of cell penetration; that luminous bacteria are cytolyzed by water, hypotonic solutions, and by freely penetrating solutions; that luminous bacteria are not injured by hydrogen or hydroxyl ions in the external solutions within the range of pH values employed with the ammonium salts and that therefore disappearance of the light in isotonic solutions of these salts must be due to penetration of the solute; and that there is a characteristic difference between the effects of strong and of weak acids and alkalies on luminous bacteria.     

光合细菌在微生物燃料电池中的应用研究进展

微生物燃料电池(Microbial fuel cells,MFCs)降解污染物的同时产生电能,受到广泛关注。光合细菌在MFCs领域的应用实现了污水处理、CO2捕捉、光电转换等多重功能,并显示出了良好的产电特性。本文根据光合细菌在MFCs中所起作用的不同对其产电机理进行评述,并在此基础上分析了光照对光合细菌型MFCs产电性能的影响;针对当前研究的不足与面临的问题,提出了今后光合细菌在MFCs领域的应用前景与发展方向。     

Influence of Sulfate-Reducing Bacteria on Outdoor Hydrogen Production by Photosynthetic Bacterium with Seawater

The application of seawater for bacterial fermentative production is a cost-effective technology. Hydrogen production by marine photosynthetic bacterium with seawater failed to continue after more than 10 days, and was accompanied by the formation of hydrogen sulfide and a change in culture color from red to black. However, substrate consumption in the blackish culture was comparable to that in a hydrogen-producing culture. A decrease in hydrogen production occurred upon the addition of sodium sulfide at concentrations of 1.5 mM or higher. PCR analysis targeted at the 16S rDNA sequence selective for sulfate-reducing bacteria revealed the existence of sulfate-reducing bacteria in inoculation cultures of the phototrophic bacterium and medium for hydrogen production. Hence, the high sulfate concentration of seawater, the low oxidation-reduction potential under hydrogen-producing conditions, and the presence of electron donors such as acetate might promote the metabolic activities of sulfate-reducing bacteria, resulting in the deterioration of hydrogen production with seawater. Received: 15 September 1999 / Accepted: 14 October 1999     

沼泽红假单胞菌乙酸光合放氢研究

依据光合细菌生长代谢特性和有机废水降解主要产物类型,11种有机物被用于沼泽红假单胞菌（Rhodopseudomonas palustris）Z菌株的光合产氢研究,其中,乙酸反应体系产氢活性最高。在此基础上,研究了该菌株的生长与产氢动力学行为,探求了影响该菌株光合放氢的主要限制性影响因素。结果表明,该菌株产氢与生长部分相关。种子培养基和菌龄对产氢活性有明显影响。细胞最适产氢和生长所需要的光照强度和温度基本一致。当种子来源于硫酸铵高菌龄预培养物或谷氨酸钠对数期预培养物时,该菌株产氢活性显著增加,产氢延滞期明显缩短。氧浓度和接种量对产氢活性也有显著影响。供氢体和氮源浓度直接决定细胞的生长与光放氢活性。在低于70 mmol/L乙酸钠和15 mmol/L谷氨酸钠时,产氢活性随底物浓度的增加而增强。谷氨酸钠浓度高于15mmol/L时,由于游离NH₄⁺的出现,产氢活性受到抑制,但却明显刺激细胞的生长。在标准状况下,该菌株的最大产氢速率可达19.4 mL·L^-1·h^-1。     

固定化技术在促进光合细菌产氢中的应用

氢气是一种新型的清洁高效能源,制氢技术的创新是目前研究的热点。将新型的技术及材料应用到生物制氢工艺中,从而促进生物制氢技术的产氢效率和工程应用是研究的重点之一。该文阐述了光合细菌在固定化生长条件下发酵产氢的最新研究进展,从固定化技术的原理、固定化方法的应用进展及影响因素几个方面进行了综述,详细阐述了包括包埋、悬浮载体附着生长及固定生物膜法等几种固定化方法对光发酵产氢的作用,介绍了国内外用于固定化的新型材料,并对今后的研究重点及方向进行了展望。     

Maximization of hydrogen production ability in high-density suspension of Rhodovulum sulfidophilum cells using intracellular poly(3-hydroxybutyrate) as sole substrate

Growth of and hydrogen production by wild-type (WT) Rhodovulum sulfidophilum were compared with those by one of its mutants lacking the poly(3-hydroxybutyrate) (PHB) biosynthesis ability (PNM2). During phototrophic growth under aerobic conditions with fixed illumination, changes in the extinction coefficient and PHB content of WT and PNM2 cells revealed interference of light penetration by PHB. WT cells synthesized PHB at an early stage of the cultivation. PHB degradation after exhaustion of acetate during the cultivation of WT resulted in a decrease of the extinction coefficient. The hydrogen production rate under anaerobic conditions with fixed illumination was examined in WT and PNM2 cell suspensions at different densities. The hydrogen production rate was determined not by the light penetration but by the kinds of hydrogen donors and the density of suspension. The highest value of the rate of hydrogen production from PHB, 33.0 ml/l/h, was improved compared with 26.6 ml/l/h, which was the highest value in hydrogen production from succinate. Under the same illumination, conversion to hydrogen from PHB is more efficient than that from succinate, which is one of the best substrates for hydrogen production. These results suggest that the hydrogen production rate can be maximized in the hydrogen production system based on PHB degradation, which is achieved in high-density suspension under external-substrate-depleted conditions after aerobic cultivation in the presence of an excess amount of acetate.     

微生物产氢研究的进展

氢能由于其清洁、高效、可再生的特点而成为一种最有吸引力的化石燃料的替代能源。与传统的热化学和电化学制氢技术相比,生物制氢具有低能耗、少污染等特点。本文主要对各种微生物的生物产氢方法作一综合概括,着重介绍光合紫色非硫细菌(PNS)产氢研究的最新进展。