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。     

Microaerobic hydrogen production by photosynthetic bacteria in a double-phase photobioreactor

The rate of hydrogen production by the marine nonsulfur photosynthetic bacterium, Rhodovulum sp., increased with increasing light intensity. A light intensity of 1800 W/m(2) hydrogen production rate was achieved at the rate of 9.4 micromol/mg dry weight/h. The hydrogen production of this strain was enhanced by the addition of a small amount of oxygen (12 micromol O(2)/reactor). Intracellular ATP content was most efficiently accumulated under microaerobic, dark conditions. Hydrogen production rate by Rhodovulum sp. was investigated using a double-phase photobioreactor consisting of light and dark compartments. This rate was compared with data obtained using a conventional photobioreactor. Rhodovulum sp. produced hydrogen at a rate of 0.38+/-0.03 micromol/mg dry weight/h under microaerobic conditions using the double-phase photobioreactor. The hydrogen production rate was four times greater under microaerobic conditions, as compared with anaerobic conditions using either type of photobioreactor. Hydrogen production using a double-phase photobioreactor was demonstrated continuously at the same rate for 150 h.     

光合细菌Rhodopseudomonas产氢的影响因子实验研究

利用光合细菌Rhodopseudomonas 8株菌株研究初期活性、光照强度、C源种类、菌株差异对生物产氢的影响表明 ,不同菌株的C源利用性有较大差异 ,但对乳酸钠都有很好的利用性 .细菌的初期活性对产氢有一定程度的影响 ,稳定生长期的细菌比对数生长期的细菌产氢活性略高 .光照强度对产氢活性的影响明显 ,在光饱和度以下 ,光照强度大则产氢速率高 .不同菌种的产氢性能有效大差异 ,从上海地区有机污染环境中分离到的RhodopseudomonasB2 1 菌株在以乳酸钠 ( 50mmol·L- 1 )为C源、谷氨酸钠 ( 1 0mol·L- 1 )为N源 ,60 0 0Lx光照、30℃下 ,最大产氢速率达到 1 4.7ml·h- 1 ·g- 1 细胞干重 .     

Sago starch as a low-cost carbon source for exopolysaccharide production by Lactobacillus kefiranofaciens

Low-cost sago starch was used as a carbon source for production of the exopolysaccharide kefiran by Lactobacillus kefiranofaciens. A simultaneous saccharification and fermentation process of sago starch for kefiran production was evaluated. Factors affecting the process such as an initial pH, temperature, starch concentration, including a mixture of α-amylase and glucoamylase were determined. The highest kefiran concentration of 0.85 g/l was obtained at the initial pH of 5.5, temperature of 30 °C, starch concentration of 4% and mixed-enzymes with activity of 100 U/g-starch. The use of a mixture of α-amylase and glucoamylase could enhance the productivity compared to the use of α-amylase alone. The optimal ratio of α-amylase to glucoamylase of 60:40 gave the highest kefiran production rate of 11.83 mg/l/h. This study showed that sago starch could serve as a low-cost substrate for kefiran production.     

Influence of nitrogen source on hydrogen generation by a photosynthetic bacterium

Axenic culture of photosynthetic purple sulphur bacterium, isolated from a paper mill effluent, grew best with ammonium chloride as nitrogen source. Cells grown with N₂ or ammonium chloride produced hydrogen with initial lag periods of 24 and 48 h, respectively. The maximum amount of hydrogen was evolved by cells grown with N₂ and with malate as electron donor.R. Lakshmi is with the Microbial Physiology Laboratory, Department of Botany, and H. Polasa is with the Department of Microbiology, Osmania University, Hyderabad 500 007, India.     

Acetate and carbon dioxide assimilation by Desulfovibrio vulgaris (Marburg), growing on hydrogen and sulfate as sole energy source

Desulfovibrio vulgaris (Marburg) was grown on hydrogen plus sulfate as sole energy source and acetate plus CO₂ as the sole carbon sources. The incorporation of U-¹⁴C acetate into alanine, aspartate, glutamate, and ribose was studied. The labelling data show that alanine is synthesized from one acetate (C-2 + C-3) and one CO₂ (C-1), aspartate from one acetate (C-2 + C-3) and two CO₂ (C-1 + C-4), glutamate from two acetate (C-1–C-4) and one CO₂ (C-5), and ribose from 1.8 acetate and 1.4 CO₂. These findings indicate that in Desulfovibrio vulgaris (Marburg) pyruvate is formed via reductive carboxylation of acetyl-CoA, oxaloacetate via carboxylation of pyruvate or phosphoenol pyruvate, and -ketoglutarate from oxaloacetate plus acetyl-CoA via citrate and isocitrate. Since C-5 of glutamate is derived from CO₂, citrate must have been formed via a (R)-citrate synthase rather than a(S)-citrate synthase. The synthesis of ribose from 1.8 mol of acetate and 1.4 mol of CO₂ excludes the operation of the Calvin cycle in this chemolithotrophically growing bacterium.     

Effects of Ethanolamine as a nitrogen source on hydrogen production by Rhodobacter capsulatus

Ethanolamine was examined as a nitrogen source in the production of hydrogen by Rhodobacter capsulatus ST-410, a hydrogenase-deficient mutant of the strain B-100. It was found that ethanolamine supports cell growth as the sole nitrogen source and permits a large amount of hydrogen evolution, detected at 138 micromol/ml-culture from 3.5 mM ethanolamine and 30 mM DL-malate. The amount corresponded to a stoichiometric yield of 77% and was close to that obtained from 7.0 mM L-glutamate and 30 mM DL-malate. The hydrogen evolution rate per unit biomass (cells) was higher than that with L-glutamate, and the cells grown with ethanolamine had higher nitrogenase activity than the cells grown with L-glutamate. In terms of bioconversion of cellulosic and hemicellulosic biomass to hydrogen, D-glucose, D-xylose, and D-cellobiose were tested as substrates. The results indicated that those sugars permit a large evolution of hydrogen through cultivation with ethanolamine as a nitrogen source. For instance, the cells grown with 3.5 mM ethanolamine evolved hydrogen of 289 micromol/ml-culture (80% yield) from 30 mM D-glucose under a controlled pH of 6.4 to 6.9.     

Temperature-sensitive PSII and promiscuous PSI as a possible solution for sustainable photosynthetic hydrogen production

Sustainable hydrogen production in cyanobacteria becomes feasible as a result of our recent studies of the structure of photosystem I encoding operon in a marine phage. We demonstrated that the fused PsaJF subunit from the phage, substituted for the two separate subunits in Synechocystis, enabled the mutated PSI to accept electrons from additional electron donors such as respiratory cytochromes. In this way, a type of photorespiration was created in which the cell consumes organic material through respiratory processes and PSI serves as a terminal electron acceptor, substituting for cytochrome oxidase. We designed a hydrogen-producing bioreactor in which this type of photorespiration could utilize the organic material of the cell as an electron source for H(2) production. We propose, in parallel, to engineer cyanobacterial and/or algal strains with a temperature-sensitive PSII and enhanced respiration rates to achieve efficient and sustainable hydrogen production. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.     

Thioacetamide as a source of hydrogen sulfide for colony growth of purple sulfur bacteria

Thioacetamide (TAA), CH₃CSNH₂, is an unstable sulfur compound which upon addition of acid decomposes into acetic acid, ammonia, and hydrogen sulfide (H₂S). This characteristic may be taken advantage of when doing plate counts or isolation streaks of phototrophic purple sulfur bacteria. We have performed plate counts and isolation streaks of these bacteria in Gas-Pak anaerobic jars (BBL) supplemented with a test tube containing 0.05 g or 0.10 g TAA dissolved in 1.0 ml of 0.2 N or 0.5 N HCl. It may be demonstrated in a Warburg respirometer that the gas is released over a period of at least 1 week. Colony growth may be observed in 5–10 days. One advantage of this technique is that sodium sulfide (Na₂S·9H₂O) need not be added to the agar medium, which, therefore, may be prepared and stored for future use. This technique has been used successfully for the isolation ofAmoebobacter, Chromatium, Ectothiorhodospira, Lamprocystis, Thiocapsa, andThiocystis.     

Protein-extracted lucerne fibres as a carbon and energy source for cellulase production by Thermomonospora curvata

Protein-extracted lucerne fibre (PELF) was evaluated as a carbon/energy source for cellulase production by the thermophilic actinomycete, Thermomonospora curvata. Shake cultures with lucerne fibre produced only one-half the cellulase obtained from growth on cotton fibre (the most inductive substrate). Control of pH at 8·0 under fermenter conditions lowered the rate of PELF carbohydrate solubilization during early growth and raised saccharifying cellulase production by about 70% to a maximum of 0·48 filter paper units/ml.     

Acetate as sole carbon and energy source for growth of methanosarcina strain 227

[This corrects the article on p. 995 in vol. 39.][This corrects the article on p. 996 in vol. 39.].     

A two-dimensional mass transfer model for an annular bioreactor using immobilized photosynthetic bacteria for hydrogen production

A two-dimensional model for substrate transfer and biodegradation in a novel, annular fiber-illuminating bioreactor (AFIBR) is proposed in which photosynthetic bacteria are immobilized on the surface of a side-glowing optical fiber to form a stable biofilm. When excited by light, the desired intensity and uniform light distribution can be obtained within the biofilm zone in bioreactor and then realize continuous hydrogen production. Substrate transfer and biodegradation within the biofilm zone, as well as substrate diffusion and convection within bulk fluid regions are considered simultaneously in this model. The validity of the model is verified experimentally. Based on the model analysis, influences of flow rate and light intensity on the substrate consumption rate and substrate degradation efficiency were investigated. The simulation results show that the optimum operational conditions for the substrate degradation within the AFIBR are: flow rate 100 ml h^?1 and light intensity 14.6 μmol photons m^?2 s^?1.     

初始底物浓度对序批式培养光合细菌产氢动力学影响

实验研究了初始底物浓度对序批式培养光合细菌生长、降解及产氢过程的影响,根据最大比生长速率实验数据拟合得到其关于初始底物浓度影响的关联式,并在建立的修正Monod模型基础上建立了光合细菌比生长速率、基质比消耗速率和比产氢速率关于底物初始浓度影响的数学模型,模型预测值与实验结果在光合细菌生长期和稳定期内得到较好吻合,反映了光合细菌生长、降解和产氢过程中受底物初始浓度限制性和抑制性影响的基本规律。分析发现光合细菌生长、降解基质和产氢过程中最适底物浓度为50 mmol/L,初始底物浓度低于或高于该浓度时,光合细菌生长、降解及产氢过程都受到限制性或抑制性影响,且抑制性影响较限制性影响效果更明显;底物比消耗速率受初始底物浓度影响较小。     

Corroding iron as a hydrogen source for sulphate reduction in growing cultures of sulphate-reducing bacteria

Summary In anaerobic corrosion experiments, hydrogenase-positiveDesulfovibrio strains, grown with limiting lactate concentrations in the presence of steel wool, formed more sulphide than expected or observed with lactate alone. The additional sulphide obviously originated from sulphate reduction with cathodically formed hydrogen from the steel surface. The hydrogenasenegativeD. sapovorans did not produce additional sulphide. The observations agree with the theory of von Wolzogen Kühr and van der Vlugt (1934) that explains anaerobic corrosion as a cathodic depolarization of iron surfaces by hydrogen-consuming sulphate-reducing bacteria. The influence of the iron surface area, the salt concentration and the pH-value on the utilization of cathodically formed hydrogen was investigated. The significance of an additional organic electron donor for the corrosion of iron in aqueous environments is discussed.