Photo-induced H2 production by [NiFe]-hydrogenase from T. roseopersicina covalently linked to a Ru(II) photosensitizer

The potential of hydrogen as a clean renewable fuel source and the finite reserves of platinum metal to be utilized in hydrogen production catalysts have provided the motivation for the development of non-noble metal-based solutions for catalytic hydrogen production. There are a number of microorganisms that possess highly efficient hydrogen production catalysts termed hydrogenases that generate hydrogen under certain metabolic conditions. Although hydrogenases occur in photosynthetic microorganisms, the oxygen sensitivity of these enzymes represents a significant barrier in directly coupling hydrogen production to oxygenic photosynthesis. To overcome this barrier, there has been considerable interest in identifying or engineering oxygen tolerant hydrogenases or generating mimetic systems that do not rely on oxygen producing photocatalysts. In this work, we demonstrate photo-induced hydrogen production from a stable [NiFe]-hydrogenase coupled to a [Ru(2,2'-bipyridine)₂(5-amino-1,10-phenanthroline)]²⁺ photocatalyst. When the Ru(II) complex is covalently attached to the hydrogenase, photocatalytic hydrogen production occurs more efficiently in the presence of a redox mediator than if the Ru(II) complex is simply present in solution. Furthermore, sustained hydrogen production occurs even in the presence of oxygen by presumably creating a local anoxic environment through the reduction of oxygen similar to what is proposed for oxygen tolerant hydrogenases. These results provide a strong proof of concept for engineering photocatalytic hydrogen production in the presence of oxygen using biohybrid mimetic systems.     

Photofermentative hydrogen production from wastes

In many respects, hydrogen is an ideal biofuel. However, practical, sustainable means of its production are presently lacking. Here we review recent efforts to apply the capacity of photosynthetic bacteria to capture solar energy and use it to drive the nearly complete conversion of substrates to hydrogen and carbon dioxide. This process, called photofermentation, has the potential capacity to use a variety of feedstocks, including the effluents of dark fermentations, leading to the development of various configurations of two-stage systems, or various industrial and agricultural waste streams rich in sugars or organic acids. The metabolic and enzymatic properties of this system are presented and the possible waste streams that might be successfully used are discussed. Recently, various immobilized systems have been developed and their advantages and disadvantages are examined.     

微生物介导反刍动物瘤胃氨生成及其对瘤胃功能的影响

反刍动物瘤胃中栖息着丰富多样的微生物,其在瘤胃内氨生成过程中发挥了重要的作用。微生物介导的氨基酸脱氨基作用和非蛋白氮水解作用是瘤胃内氨生成的主要途径。微生物介导了瘤胃内氨的生成,同时瘤胃内产生的氨也会反馈影响微生物菌群结构及瘤胃上皮功能,进而影响瘤胃发酵及宿主健康。本文主要综述了瘤胃微生物在介导氨生成中的作用和氨对瘤胃消化及瘤胃上皮功能的影响,以期对后续研究有所启发。     

Two-step immobilized enzyme conversion of cephalosporin C to 7-aminocephalosporanic acid

The first large-scale production of 7-aminocephalosporanic acid (7ACA) from cephalosporin C (CPC) using a wholly enzymatic synthesis method is reported here. We produced 7ACA from CPC in as high a molar yield as 85% using the immobilized enzymes D-amino acid oxidase (D-AOD) and glutaryl-7-ACA acylase (GL-acylase). In the first reactor, CPC is converted to keto-adipyl-7-aminocephalosporanic acid (keto-7ACA) using an immobilized D-AOD isolated from a yeast, Trigonopsis variabilis. The keto-7ACA is then spontaneously converted to glutaryl-7-aminocephalosporanic acid (GL-7ACA) via a chemical reaction with hydrogen peroxide. The hydrogen peroxide is also a product of the D-AOD reaction. Near quantitative conversion of the keto-7ACA to GL-7ACA was observed. The second reactor converts GL-7ACA to 7ACA using an immobilized GL-acylase, which was isolated from a reconbinant Escherichia coli. The final 7ACA crystalline product is a high quality product. The reactions are conducted under very mild aqueous conditions: pH 8.0 and 20 degrees to 25 degrees C. The production of desacetyl side products is minimal. This process is currently being implemented on an industrial scale to produce 7ACA. (c) 1995 John Wiley & Sons, Inc.     

固定化微生物处理氨氮废水的研究进展

生物方法因其高效、持久及无二次污染已经成为治理水污染的重要方法,固定化微生物不仅加强微生物治理效果,更能保证微生物生存稳定,是现阶段生物方法治理水污染的主要组分。近年来学者们也做了很多固定化微生物处理各类废水的研究,并取得了大量优异成果。文章对固定化微生物技术进行剖析,分析各类固定化载体材料及各种固定化方法优缺点;解析利用固定化微生物处理氨氮废水效果研究及固定化微生物处理效果影响因素;综述近年来固定化微生物处理氨氮废水应用情况;最后进行总结,并提出自己的展望。     

Production of molecular hydrogen with immobilized cells ofRhodospirillum rubrum

Summary Cells ofRhodospirillum rubrum have been immobilized in various gels and tested for photobiological hydrogen production. Agar proved to be the best immobilizing agent with respect to production rates as well as stability. Agar immobilized cells were also superior compared to liquid suspension cultures. Growth conditions of the cells prior to immobilization, e.g. cell age, light intensity or nutrient composition, were of primary importance for the activity in the later immobilized state. A reactor with agar immobilized cells has been operated successfully over 3000 h with a loss of the activity of about 60%. Mean rates for hydrogen production for immobilized cells in this work during the first 60 to 70 hours after immobilization were in the range of 18 to 34 μl H₂ mg⁻¹ d.w. h⁻¹ and thus by a factor of up to 2 higher than liquid cultures under the same conditions. Maximal rates of hydrogen production (57 μl H₂ ml⁻¹ immobilized cell suspension) were reached in agar gel beads with cells immobilized after 70 h growth in liquid culture in the light and a cell density of 1.0 mg ml⁻¹, 70 h after immobilization.     

Bioengineering aspects of biophotolysis

The practical aspects of producing hydrogen by photosynthetic microorganisms are reviewed. Various alternative concepts for hydrogen production are discussed, both single and two-stage systems. The best developed process currently is based on nitrogen-fixing heterocystous blue-green algae which can produce hydrogen and oxygen simultaneously. Solar energy conversion to hydrogen efficiencies of 0.2% averaged over several weeks have been obtained with outdoor systems. Practical systems would require a ten-fold increase in conversion efficiencies. Also, systems which produce pure hydrogen are preferred. Photosynthetic bacteria are of near-term applications. A general design for a biophotolysis system is proposed consisting of vertically arranged, thin-walled glass tubes with an inert gas recirculated through the cultures for mixing and removal of hydrogen. Gas mass transfer considerations, energy utilization, and economics favour such a system.     

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。     

A chemiluminescence automatic analyser for the measurement of biological compounds

A compact automated analyser which could analyse constituents in biological fluids with a small sample volume and in a short time has been developed. The instrument was composed of a flow injection analysis system equipped with chemiluminometric detection and an immobilized enzyme column reactor used in combination. Chemiluminescence has high sensitivity, and its reaction proceeds very quickly. Furthermore, an immobilized enzyme column reactor can produce a sufficient amount of hydrogen peroxide from compounds in serum in a short time. When enzymes are used as reagents for the analysis of substances in blood or blood serum, the final signals emitted by different enzyme reactions are usually not only hydrogen peroxide but also ammonia, NAD(P)H and so on. However, the practical chemiluminescence method for ammonia and NAD(P)H has not been established. We have discovered a new practical method for ammonia and NAD(P)H using an enzyme column reactor consisting of both immobilized L -glutamate dehydrogenase and L -glutamate oxidase. The determinations of glucose and uric acid in serum by chemiluminometry after production of hydrogen peroxide by the respective oxidases are presented. A newly chemiluminometric determination of ammonia, NAD(P)H and its applications to other enzymatic analyses that give ammonia and NAD(P)H as a final signal are also described.     

Biofiltration--the treatment of fluids by microorganisms immobilized into the filter bedding material: a review

Biofiltration is distinguished from other biological waste treatments by the fact that there is a separation between the microorganisms and the treated waste. In biofiltration systems the microorganisms are immobilized to the bedding material, while the treated fluid flows through it. In recent decades, a vast amount of literature has been written on single experiments involving the treatment of fluids by immobilized microorganisms. Several artificial immobilization methods have been examined and impressive results have been achieved in the treatment of fluids with one of the artificial immobilization methods the entrapment of microorganisms within polymer beads. This method, even though it needs to be improved, seems to have a future potential in commercial biofiltration systems. The methods of artificial immobilization of microorganisms within biofiltration systems have several advantages, but also suffer from several disadvantages in comparison to the treatment of fluids by naturally attached microorganisms. Understanding the mechanisms and forces responsible for the attachment of microbes to the bedding material, in attempt to improve this attachment, is of the utmost importance. Further improvement of the artificial entrapment of microorganisms within polymers will allow the exploitation of the advantages of this method in the treatment of fluids. The aim of this review essay is to introduce the main principles of two immobilization processes - the self-attachment of microorganisms to the bedding material and the artificial entrapment of microorganisms within polymer beads. Both treatments of liquids and gases with each immobilization process are discussed. The advantages and disadvantages of each immobilization process are pointed out and different aspects of the fluid treatment with the two immobilization processes are compared.     

微生物发酵产氢的影响因素分析*

随着环保要求的愈益严格和化石能源的日益短缺,氢作为清洁高效的可再生能源日益受到人们的重视。微生物发酵产氢可以利用可再生的生物质,符合可持续发展的要求。针对影响微生物发酵产氢的因素,总结了国内外在该领域的研究成果,重点介绍了产氢微生物、营养物、产物和工艺操作条件等方面对发酵产氢的影响,同时还阐述了以有机废弃物为基质时的发酵产氢影响因素。     

Ammonia removal from livestock wastewater by ammonia-assimilating microorganisms immobilized in polyvinyl alcohol

We isolated ammonia-assimilating microorganisms from the livestock manure treatment systems and evaluated their ammonia-assimilating ability. Many isolates utilized ammonia at high rates when they were purely cultivated in a nitrogen-limited medium to which sterilized lagoon extract had been added. Some isolates that were immobilized in polyvinyl alcohol (PVA) utilized ammonia present in the media containing viable lagoon microorganisms. Staining with 4′,6′-diamidino-2-phenylindole (DAPI) indicated that the immobilized high ammonia-assimilating isolates grew dominantly within the PVA beads. High ammonia-assimilating isolates in the mixed culture containing viable lagoon microorganisms were identified as Pseudomonas spp. and member of Rhizobiaceae species by partial sequencing of the 16S ribosomal DNA.     

Immobilized cell technology applied in solubilization of insoluble inorganic (rock) phosphates and P plant acquisition

This paper reviews current knowledge of the production of organic acids by immobilized microorganisms with a simultaneous solubilization of rock phosphate in fermentation and soil conditions. The most widely applied methods are based on the passive immobilization in preformed porous carriers and entrapment of the microbial cells in natural gels. In general, immobilized systems show higher acid producing and rock phosphate solubilizing activity than freely suspended cells. The potential of gel-entrapped P-solubilizers and mycorrhizal fungi as microbial soil inoculants is also pointed out. Some advantages and constraints of using immobilized cells are discussed and a special emphasis on further research is given.     

Use of a tapered fluidized bed as a Continuous bioreactor

Reactor systems based on tapered fluidized beds are being developed for aqueous bioprocesses in which adhering microorganisms or immobilized active biological fractions are used. The use of a fluidized bed prevents biomass buildup, accommodates particulates in the feed stream, is compatible with gas sparging, and allows easy removal or addition of the active materials. The tapered reactor tends to stabilize the fluidized bed, thus allowing a much wider range of operating conditions. Preliminary experimental results and an empirical mathematical model of the tapered bed indicate that bed stability is associated with a decreasing velocity and void-fraction profile up the bed and the pressure drop across the bed decreases with increasing flow rates. The tapered fluidized bed bioreactor is being evaluated for use in the enzymatic production of hydrogen, microbiological denitrification, and microbiological degradation of coal conversion aqueous waste streams. The enzyme catalyzed conversion of lactose to glucose and galactose was used in the evaluation of the reactor concept.     

Biosorption and recycling of gold using various microorganisms

In order to obtain basic information on the biosorption and recycling of gold from aqueous systems using microbial cells, the biosorption of gold by various microorganisms was investigated. Of 75 strains of microorganisms tested (25 bacteria, 19 actinomycetes, 17 fungi and 14 yeasts), high abilities of gold biosorption from a solution containing hydrogen tetrachloroaurate (III) were found in some gram-negative bacterial strains, such as Acinetobacter calcoaceticus, Erwinia herbicola, Pseudomonas aeruginosa, and P. maltophilia. Most of the gram-positive bacteria, actinomycetes, fungi and yeasts had a lower ability for gold biosorption than gram-negative bacteria. On the other hand, all of the microorganisms tested adsorbed far smaller amounts of gold from a solution containing gold dicyanoaurate (I). The biosorption of gold from a solution containing hydrogen tetrachloroaurate (III) using P. maltophilia having a high adsorbing ability for gold was very rapid and was affected by the pH of the solution, external gold concentration, and cell amounts. P. maltophilia cells immobilized with polyacrylamide gel also have a high ability for gold biosorption. The gold adsorbed on the immobilized cells is easily desorbed with 0.1 M thiourea solution. The immobilized P. maltophilia cells can be used repeatedly in biosorption-desorption cycles.     

Immobilized plant cell reactors

The use of plant cells for the production of biochemicals represents a new area of biotechnological exploration. The techniques envisioned for industrial processes are related to those developed for microorganisms and a strong emphasis should be placed on immobilized cell systems. This review examines the spectrum of products that are synthesized by higher plants and the immobilization techniques that are suited to entrap plant cells from suspension culture. Different reactor configurations are described. Both packed-bed reactors with alginate-entrapped cells and hollow-fibre cartridges with sequestered cells have utility for the continuous production of biochemicals.     

Microbiology of synthesis gas fermentation for biofuel production

A significant portion of biomass sources like straw and wood is poorly degradable and cannot be converted to biofuels by microorganisms. The gasification of this waste material to produce synthesis gas (or syngas) could offer a solution to this problem, as microorganisms that convert CO and H2) (the essential components of syngas) to multicarbon compounds are available. These are predominantly mesophilic microorganisms that produce short-chain fatty acids and alcohols from CO and H2. Additionally, hydrogen can be produced by carboxydotrophic hydrogenogenic bacteria that convert CO and H2O to H2 and CO2. The production of ethanol through syngas fermentation is already available as a commercial process. The use of thermophilic microorganisms for these processes could offer some advantages; however, to date, few thermophiles are known that grow well on syngas and produce organic compounds. The identification of new isolates that would broaden the product range of syngas fermentations is desirable. Metabolic engineering could be employed to broaden the variety of available products, although genetic tools for such engineering are currently unavailable. Nevertheless, syngas fermenting microorganisms possess advantageous characteristics for biofuel production and hold potential for future engineering efforts.     

Effect of ammonia concentration on fermentative hydrogen production by mixed cultures

The effect of ammonia concentration ranging from 0 to 10 g N/L on fermentative hydrogen production by mixed cultures was investigated in batch tests using glucose as substrate at 35 degrees C and initial pH 7.0. The experimental results showed that during the fermentative hydrogen production, the substrate degradation efficiency increased with increasing ammonia concentration from 0 to 0.01 g N/L. The hydrogen production potential, hydrogen yield and average hydrogen production rate increased with increasing ammonia concentration from 0 to 0.1g N/L. The maximum hydrogen production potential of 291.4 mL, maximum hydrogen yield of 298.8 mL/g glucose and maximum average hydrogen production rate of 8.5 mL/h were all obtained at the ammonia concentration of 0.1g N/L.     

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.     

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。