Lipoamide dehyrogenase immobilized on porous glass.

Lipoamide dehydrogenase (NADH:lipoamide oxidoreductase, EC 1.6.4.3) isolate from pig heart and Escherichia coli was covalently coupled by both diazonium and amide bonds to controlled pore glass beads (96% silica). When the enzyme was immobilized in the presence of NAD+, the enzyme no longer exhibited its normal requirement for NAD+ for full activity. If the immobilized enzyme was then treated with NADase, the requirement for NAD+ was restored. Enzyme immobilized in the absence of NAD+ exhibited normal NAD+ dependence both prior to an after NADase treatment. These results are discussed in terms of co-immobilization of NAD+ at or near the allosteric site of the enzyme.     

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.     

Properties of ribulose diphosphate carboxylase immobilized on porous glass

Ribulose-1,5-diphosphate carboxylase from spinach has been bound to arylamine porous glass with a diazo linkage and to alklamine porous glass with glutaraldehyde. Stability at elevated temperatures and responses to changes of pH and ribulose-1,5-diphosphate, Mg²⁺, and dithiothreitol concentrations were not significantly different from the soluble enzyme, though stability at 4°C was somewhat improved.     

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。     

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 .     

Atmospheric methane removal by methane-oxidizing bacteria immobilized on porous building materials

Biological treatment using methane-oxidizing bacteria (MOB) immobilized on six porous carrier materials have been used to mitigate methane emission. Experiments were performed with different MOB inoculated in building materials at high (~20 % (v/v)) and low (~100 ppmv) methane mixing ratios. Methylocystis parvus in autoclaved aerated concrete (AAC) exhibited the highest methane removal rate at high (28.5?±?3.8 μg CH₄ g^?1 building material h^?1) and low (1.7?±?0.4 μg CH₄ g^?1 building material h^?1) methane mixing ratio. Due to the higher volume of pores with diameter >5 μm compared to other materials tested, AAC was able to adsorb more bacteria which might explain for the higher methane removal observed. The total methane and carbon dioxide-carbon in the headspace was decreased for 65.2?±?10.9 % when M. parvus in Ytong was incubated for 100 h. This study showed that immobilized MOB on building materials could be used to remove methane from the air and also act as carbon sink.     

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.     

H production by immobilized cells of Clostridium butyricum on porous glass beads

Clostridium butyricum immobilized on porous glass beads in a column reactor evolved H 2 at 715 and 1,150 ml/l.h, with H 2 yields of 2.3 and 1.9 mol H 2 /mol glucose, at retention times of 2.0 and 1.0 h, respectively, with a medium containing 0.5 g glucose/l in continuous cultures without pH control.     

The properties of subtilisin,Novo type,immobilized on porous glass

Studies of the properties of subtilisin, Novo type, immobilized on porous glass with the aid of hexamethylene diisocyanate were carried out. The immobilized proteinase preparation shows optimum activity at a pH value of 10.7 and at a temperature between 60–65°C. It was stable in a wider range of pH and temperature values than the native subtilisin. The K_M values for hemoglobin and BAEE were 9.2 × 10^?5 [M] and 139 × 10^?5 [M], respectively. Under relatively non-aqueous conditions, immobilised subtilisin was able to synthesize phenylacetic acid ethyl ester.     

Nitrogen fixation and hydrogen metabolism in photosynthetic bacteria.

The photosynthetic bacteria are found in a wide range of specialized aquatic environments. These bacteria represent important members of the microbial community since they are capable of carrying out two of the most important processes on earth, namely, photosynthesis and nitrogen fixation, at the expense of solar energy. Since the discovery that these bacteria could fix atmospheric nitrogen, there has been an intensification of studies relating to both the biochemistry and physiology of this process. The practical importance of this field is emphasized by a consideration of the tremendous energy input required for the production of artificial nitrogenous fertilizer. The present communication aims to briefly review the current state of knowledge relating to certain aspects of nitrogen fixation by the photosynthetic bacteria. The topics that will be discussed include a general survey of the nitrogenase system in the various photosynthetic bacteria, the regulation of both nitrogenase biosynthesis and activity, recent advances in the genetics of the nitrogen fixing system, and the hydrogen cycle in these bacteria. In addition, a brief discussion of some of some of the possible practical applications provided by the photosynthetic bacteria will be presented.     

Affinity chromatography of proteinases using bacitracin immobilized to porous glass beads

J. FONTECHA, T. REQUENA AND H.E. SWAISGOOD. 1996. This study describes an affinity chromatography procedure for proteinase purification using bioselective binding to immobilized bacitracin. By coupling bacitracin to controlled-pore glass (CPG) beads, an affinity matrix was obtained that permitted rapid purification of proteinases under conditions that minimize autolysis. Bacitracin-CPG was used to bioselectively adsorb the extracellular proteinase secreted by Enterococcus faecalis var. liquefaciens IFPL 383. The overall purification obtained with this procedure was 5149-fold. The ability of bacitracin-CPG to bind other proteinases was examined using various commercial proteinases. The specific activities of subtilin BPN' and proteinase K were increased by bioselective adsorption and excellent recoveries of all proteinases applied were obtained.     

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.     

Some observations on immobilized hydrogen-producing bacteria: Behavior of hydrogen in gel membranes

A hydrogen-producing bacterium, Clostridium butyricum, was immobilized in polyacrylamide gel membrane, agar gel membrane, and collagen membrane. The apparatus consisted of a compartment(I or II) and a whole-cell-entrapped gel membrane which was placed in the center of the two compartments. The behavior of hydrogen through whole-cell-entrapped gel membrane was examined for improving the sensitivity of a microbial BOD sensor and the efficiency of biochemical fuel cells. The bacteria-polyacrylamide gel membrane produced a higher amount of hydrogen in compartment II than the other bacteria-entrapped gel membranes. The apparent diffusion constant decreased with increasing bacteria contents in the gel membrane. As a result, 100 mg wet cells/g gel was the maximal bacteria content in the membrane for the production of hydrogen. Twenty percent of the hydrogen evolved by the whole cells diffused to compartment II, and there was a linear relationship between the glucose concentration in a bulk solution (compartment I) and the rate of hydrogen diffused to compartment II.     

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     

Relationship between the photoproduction of hydrogen and the accumulation of PHB in non-sulphur purple bacteria

Wild type cells of Rhodobacter sphaeroides and Rhodospirillum rubrum strains Ha and S1 as well as mutant cells defective in the synthesis of poly-(3-Hydroxybutyric acid) (PHB), were used to study the competition between PHB accumulation and photoproduction of hydrogen for reducing equivalents. Mutants were isolated after transposon (Tn5) or N-methyl-N-nitro-N-nitrosoguanidine mutagenesis. The PHB-defective mutants of R. sphaeroides lacked PHB synthase activity. In two mutants Tn5 was inserted in the PHB synthase gene. No mutants occured that lacked the activity of -ketothiolase or acetoacetyl-coenzyme A reductase. Pronounced competitive effects occured only with acetate as the organic substrate. With other organic acids or sugars, which are less readily converted to PHB than acetate, competitive effects were not significant or absent. Correspondence to: H. G. Schlegel     

Pore diffusion model for a two-substrate enzymatic reaction: Application to galactose oxidase immobilized on porous glass particles

An analysis of the pore diffusion model involving a two-substrate enzymatic reaction is presented. The resulting equations have been applied to the case of galactose oxidase catalyzed oxidation of galactose when the enzyme is immobilized on porous glass particles. The physical constants of the system were obtained by theoretical predictions and the enzyme concentration in the porous medium was derived from the experimental results. The calculations were performed with the assumption that the kinetic parameters of the enzyme remain unchanged upon immobilization. The theoretically calculated effectiveness factors were compared with the experimental effectiveness factors determined from the batch kinetic experiments and were found to be in agreement. The results are presented as effectiveness factor plots graphed as functions of bulk galactose and oxygen concentrations. The model was extended in order to study the effect of external mass transfer coefficients and pore enzyme concentrations on the effectiveness factors.     

Aerobic anoxygenic photosynthetic bacteria with zinc-bacteriochlorophyll

Naturally occurring chlorophyllous pigments, which function as the cofactor in the early photochemical reaction of photosynthesis, have been proven beyond question to be magnesium-complexed porphyrin derivatives. Phototrophic organisms that use (bacterio)chlorophylls ([B]Chls) containing metals other than Mg were unknown for a long time. This common knowledge of natural photosynthesis has recently been modified by the striking finding that a novel purple pigment, zinc-chelated-BChl (Zn-BChl) a, is present as the major and functional pigment in species of the genus Acidiphilium. Acidiphilium species are obligately acidophilic chemoorganotrophic bacteria that grow and produce photopigments only under aerobic conditions. Although the mechanism of photosynthesis with Zn-BChl a in Acidiphilium species is similar to that seen in common purple bacteria, some characteristic photosynthetic features of the acidophilic bacteria are also found. The discovery of natural photosynthesis with Zn-BChl has not only provided a new insight into our understanding of bacterial photosynthesis but also raised some interesting questions to be clarified. The major questions are why the acidophilic bacteria have selected Zn-BChl for their photosynthesis and how they synthesize Zn-BChl and express photosynthetic activity with it in their natural habitats. In this article we review the current knowledge of the biology of Acidiphilium as aerobic photosynthetic bacteria with Zn-BChl a and discuss the interesting topics noted above.