Optimal conditions for the production of sulfated polysaccharide by marine microalga Gyrodinium impudicum strain KG03

A marine microalga Gyrodinium impudicum strain KG03 produced sulfated exopolysaccharide designated as p-KG03, which showed a strong antiviral activity against encephalomyocarditis virus (EMCV). To optimize culture conditions for the production of p-KG03, mineral salts, vitamins, plant growth hormones, temperature, pH and light conditions were examined. From this study, M-KG03 medium for the maximum production of p-KG03 was suggested as follows; NH(4)Cl 75 microM, NaH(3)PO(4) 200 microM, NaHCO(3) 50 microM, Na(2)SO(4) 10 microM, FeCl(2) x 6H(2)O 10 microM, MnCl(2) x 4H(2)O 0.1 microM, vitamin B(12) 0.75 microg, naphthalene acetic acid (NAA) 7.5 microg and myo-inositol 200 mg per liter of aged sea water. The optimal temperature and pH were 22.5 degrees C and 8.0, respectively. The optimal light conditions of intensity and period were 150 microE m(-2) s(-1) and 16:8 h light:dark cycle. Finally, the cell growth and p-KG03 production were measured in one liter of M-KG03 medium with 1% CO(2) and 50 ml min(-1) of airflow using two liters airlift balloon type photobioreactor (ABTPR). At these optimal conditions, p-KG03 production and cell growth were 134.6+/-5.9 mg l(-1) and 123,076+/-1,597 cells ml(-1), respectively, representing a 7.7 and 5.1 times compared with f/2 medium with Erlenmeyer flask culture (p-KG03 production 17.5+/-1.3 mg l(-1) and cell growth 24,311+/-1,291 cells ml(-1)).     

The Calvin cycle in cyanobacteria is regulated by CP12 via the NAD(H)/NADP(H) ratio under light/dark conditions

In Synechococcus PCC7942 cells grown in the dark, the concentrations of NAD(H) and NADP(H) were 128+/-2.5 and 483+/-4.0 microm, respectively, while those in the cells under light conditions were 100+/-5.0 and 649+/-7.0 microm, respectively. Analysis of gel filtration indicated that the change of the ratio of NADP(H) to NAD(H) in cyanobacterial cells under light/dark conditions controls the reversible dissociation of the PRK/CP12/GAPDH complex (approximately 520 kDa) consisting of phosphoribulokinase (PRK), CP12, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). S. 7942 CP12 lacked the two Cys residues essential for formation of the N-terminal peptide loop in the CP12 of higher plants, but the N-terminal region of S. 7942 CP12 had the ability to be associated with PRK. The growth of mutant cells in which the CP12 gene was disrupted by a kanamycin resistance cartridge gene was almost the same as that of wild-type cells under continuous light conditions. However, under the light/dark cycle (12 h/12 h), the growth of CP12-disrupted mutant cells was significantly inhibited compared with that of wild-type cells. The mutant cells showed a decreased rate of O2 consumption and an increased level of ribulose 1,5-bisphosphate compared with wild-type cells in the dark. These data suggest that under light and dark conditions, the oligomerization of CP12 with PRK and GAPDH regulates the activities of both enzymes and thus the carbon flow from the Calvin cycle to the oxidative pentose phosphate cycle.     

Regulation of growth and photosynthesis by Oscillatoria agardhii grown with a light/dark cycle

Abstract The cyanobacterium Oscillatoria agardhii was grown in turbidostat cultures with the light energy supply in either the continuous mode or in the pulsed mode (8/16 h light/dark (L/D) cycle). The light irradiance value used was sufficient to allow the maximal growth rate to be attained, when supplied continuously. Adaptation of O. agardhii to the L/D cycle was characterized by an increase in pigment content and photosynthetic performance, accompanied by a decrease in growth rate. This mode of adaptation resembled the adaptation of O. agardhii to continuous low light intensities. It is suggested that in this case the L/D cycle provokes this adaptation in order to allow the cells to accumulate carbohydrate rapidly during the light period. This was attributed to the storage of polyglucose, which served as a carbon and energy source for growth in the dark. The utilization of polyglucose in the dark was able to sustain the synthesis of all other cell components at the same rate as when cells were growing in the light. The growth yield in the dark, whilst metabolizing internally stored polyglucose, was 0.52 g cell C/g polyglucose C, or 0.62 g cell dry weight/g polyglucose. Although in the pulsed mode there is a 66% loss in light irradiance per 24 h when compared with a continuous light regime, the growth rate of the cyanobacteria grown in the pulsed mode was only 35% lower than the growth rate of a culture grown in continuous light. This can be explained by a high growth yield in the dark and by increased CO₂ fixation rates in the light of cells grown in the pulsed mode.     

Light inhibition of mitochondrial respiration in a mutant of Chlamydomonas reinhardtii devoid of ribulose-1,5-bisphosphate carboxylase/oxygenase activity

The effect of light on mitochondrial respiration has been investigated in Chlamydomonas reinhardtii rcl-u-1-10-6C, a mutant devoid of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity. No CO2 uptake was observed in the light, confirming that there was no Rubisco activity, but the CO2 evolution rate was diminished by 65 to 80%. This inhibition was ascribable to a decrease in the tricarboxylic acid cycle (Krebs cycle) activity. At the same time, O2 evolution associated with stimulation of the O2 uptake appears. Darkness or addition of DCMU fully reversed the effect of light, indicating that the inhibitory process is linked to photosystem activities. Levels of pyridine nucleotides (NAD(H) and NADP(H)) and adenine nucleotides (ATP and ADP), the most probable mediators of the interaction between photosynthesis and respiration, were measured in dark and in light. During a dark to light transition the level of NADPH increased significantly whereas the NAD(H) pool remained almost fully oxidized. The level of ADP was always extremely low. These results suggest that the inhibition of Krebs cycle activity is due to a competition for cytosolic ADP between chloroplastic photophosphorylations and oxidative phosphorylations.     

Sustained photobiological hydrogen gas production upon reversible inactivation of oxygen evolution in the green alga Chlamydomonas reinhardtii

The work describes a novel approach for sustained photobiological production of H(2) gas via the reversible hydrogenase pathway in the green alga Chlamydomonas reinhardtii. This single-organism, two-stage H(2) production method circumvents the severe O(2) sensitivity of the reversible hydrogenase by temporally separating photosynthetic O(2) evolution and carbon accumulation (stage 1) from the consumption of cellular metabolites and concomitant H(2) production (stage 2). A transition from stage 1 to stage 2 was effected upon S deprivation of the culture, which reversibly inactivated photosystem II (PSII) and O(2) evolution. Under these conditions, oxidative respiration by the cells in the light depleted O(2) and caused anaerobiosis in the culture, which was necessary and sufficient for the induction of the reversible hydrogenase. Subsequently, sustained cellular H(2) gas production was observed in the light but not in the dark. The mechanism of H(2) production entailed protein consumption and electron transport from endogenous substrate to the cytochrome b(6)-f and PSI complexes in the chloroplast thylakoids. Light absorption by PSI was required for H(2) evolution, suggesting that photoreduction of ferredoxin is followed by electron donation to the reversible hydrogenase. The latter catalyzes the reduction of protons to molecular H(2) in the chloroplast stroma.     

The light/dark cycle operation with an hour-scale period enhances caffeine production by Coffea arabica cells

The intermittent light irradiation with an hour-scale period is used for producing caffeine by Coffea arabica cells. Three factors concerning the light/dark cycle operation such as light intensity, the length of the cycle (period), and the ratio of the illumination time to the dark time (light/dark ratio) were investigated to optimize the caffeine production efficiency regarding light consumption. The light/dark ratio of 1/1 enhanced caffeine production, reaching the same level as continuous light; thus, the intermittent light irradiation improved the production efficiency twofold. The production was not influenced by the period, but was determined by light intensity regardless of intermittent or continuous light irradiation.     

Optimization of metabolic capacity and flux through environmental cues to maximize hydrogen production by the cyanobacterium "Arthrospira (Spirulina) maxima"

Environmental and nutritional conditions that optimize the yield of hydrogen (H(2)) from water using a two-step photosynthesis/fermentation (P/F) process are reported for the hypercarbonate-requiring cyanobacterium "Arthrospira maxima." Our observations lead to four main conclusions broadly applicable to fermentative H(2) production by bacteria: (i) anaerobic H(2) production in the dark from whole cells catalyzed by a bidirectional [NiFe] hydrogenase is demonstrated to occur in two temporal phases involving two distinct metabolic processes that are linked to prior light-dependent production of NADPH (photosynthetic) and dark/anaerobic production of NADH (fermentative), respectively; (ii) H(2) evolution from these reductants represents a major pathway for energy production (ATP) during fermentation by regenerating NAD(+) essential for glycolysis of glycogen and catabolism of other substrates; (iii) nitrate removal during fermentative H(2) evolution is shown to produce an immediate and large stimulation of H(2), as nitrate is a competing substrate for consumption of NAD(P)H, which is distinct from its slower effect of stimulating glycogen accumulation; (iv) environmental and nutritional conditions that increase anaerobic ATP production, prior glycogen accumulation (in the light), and the intracellular reduction potential (NADH/NAD(+) ratio) are shown to be the key variables for elevating H(2) evolution. Optimization of these conditions and culture age increases the H(2) yield from a single P/F cycle using concentrated cells to 36 ml of H(2)/g (dry weight) and a maximum 18% H(2) in the headspace. H(2) yield was found to be limited by the hydrogenase-mediated H(2) uptake reaction.     

Reactive oxygen species in programmed death of pea guard cells

Hydrogen peroxide potentiates CN(-)-induced apoptosis of guard cells recorded as destruction of cell nuclei in the epidermis from pea leaves. A still stronger effect was exerted by the addition of H2O2 and NADH, which are the substrates of the plant cell wall peroxidase producing O2*- coupled to the oxidation of NADH. The CN(-)-or (CN(-) + H2O2)-induced destruction of guard cell nuclei was completely removed by nitroblue tetrazolium (NBT) oxidizing O2*- and preventing there-by the subsequent generation of H2O2. The reduced NBT was deposited in the cells as formazan crystals. Cyanide-induced apoptosis was diminished by mannitol and ethanol, which are OH* traps. The dyes Rose Bengal (RB) and tetramethylrhodamine ethyl ester (TMRE) photosensitizing singlet oxygen production suppressed the CN(-)-induced destruction of the cell nuclei in the light. This suppression was removed by exogenous NADH, which reacts with 1O2 yielding O2*-. Incubation of leaf slices with RB in the light lowered the photosynthetic O2 evolution rate and induced the permeability of guard cells for propidium iodide, which cannot pass across intact membranes. Inhibition of photosynthetic O2 evolution by 3-(3',4'-dichlorophenyl)-1,1-dimethylurea or bromoxynil prevented CN(-)-induced apoptosis of guard cells in the light but not in the dark. RB in combination with exogenous NADH caused H2O2 production that was sensitive to NBT and estimated from dichlorofluorescein (DCF) fluorescence. Data on NBT reduction and DCF and TMRE fluorescence obtained using a confocal microscope and data on the NADH-dependent H2O2 production are indicative of generation of reactive oxygen species in the chloroplasts, mitochondria, and nuclear region of guard cells as well as with participation of apoplastic peroxidase. Cyanide inhibited generation of reactive oxygen species in mitochondria and induced their generation in chloroplasts. The results show that H2O2, OH*, and O2*- resources utilized for H2O2 production are involved in apoptosis of guard cells. It is likely that singlet oxygen generated by RB in the light, judging from the permeability of the plasmatic membrane for propidium iodide, makes Photosystem II of chloroplasts inoperative and induces necrosis of the guard cells.     

Kinetic modeling of light limitation and sulfur deprivation effects in the induction of hydrogen production with Chlamydomonas reinhardtii: Part I. Model development and parameter identification

Chlamydomonas reinhardtii is a green microalga capable of turning its metabolism towards H2 production under specific conditions. However this H2 production, narrowly linked to the photosynthetic process, results from complex metabolic reactions highly dependent on the environmental conditions of the cells. A kinetic model has been developed to relate culture evolution from standard photosynthetic growth to H2 producing cells. It represents transition in sulfur-deprived conditions, known to lead to H2 production in Chlamydomonas reinhardtii, and the two main processes then induced which are an over-accumulation of intracellular starch and a progressive reduction of PSII activity for anoxia achievement. Because these phenomena are directly linked to the photosynthetic growth, two kinetic models were associated, the first (one) introducing light dependency (Haldane type model associated to a radiative light transfer model), the second (one) making growth a function of available sulfur amount under extracellular and intracellular forms (Droop formulation). The model parameters identification was realized from experimental data obtained with especially designed experiments and a sensitivity analysis of the model to its parameters was also conducted. Model behavior was finally studied showing interdependency between light transfer conditions, photosynthetic growth, sulfate uptake, photosynthetic activity and O2 release, during transition from oxygenic growth to anoxic H2 production conditions.     

Effect of some environmental factors on cyanophage AS-1 development in Anacystis nidulans

The development cycle of the cyanophage AS-1 was studied in the host blue-green alga, Anacystis nidulans, under conditions that impair photosynthesis and under various light/dark regimes. Under standard conditions of incubation the 16-h development cycle consisted of a 5-h eclipse period and an 8-h latent period. Burst size was decreased by dark incubation to 2% of that observed in the light. An inhibitor of photosystem II, 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU), reduced the burst size to 27% of that of the uninhibited control, whereas cyanophage production was completely abolished by carbonyl-cyanide m-chlorophenyl hydrazone (CCCP), an inhibitor of photosynthetic electron transport. Dark incubation of infected cells decreased the latent period by 1–2 h and the eclipse period by 1 h, once the cultures were illuminated. This suggests that adsorption took place in the dark. Intracellular growth curves indicated that light is necessary for viral development. Infected cells must be illuminated at least 13 h to produce a complete burst at the same rate as the continuously illuminated control. Low light intensities retarded the development cycle, and at lowest light intensities no phage yield was obtained. AS-1 is highly dependent on host cell photophosphorylation for its development.List of Abbreviations CCCP Carbonyl-cyanide m-chlorophenyl hydrazone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl urea - m.o.i. multiplicity of infection - O.D. optical density - PFU plaque-forming unit Dedicated to Prof. Roger Y. Stanier on the occasion of his 60th birthday     

Hydrogen production by Anabaena variabilis PK84 under simulated outdoor conditions

Hydrogen production by autotrophic, vanadium-grown cells of Anabaena variabilis PK84, a cyanobacterial mutant impaired in the utilization of molecular hydrogen, has been studied under simulated outdoor conditions. The cyanobacterium was cultivated in an automated helical tubular photobioreactor (4.35 L) under air containing 2% CO(2), with alternating 12-h light (36 degrees C) and 12-h dark (14 degrees to 30 degrees C) periods. A. variabilis steadily produced H(2) directly in the photobioreactor during continuous cultivation for 2.5 months. The maximum H(2) production by the continuously aerated culture under light of 332 microE. s(-1). m(-2) was 230 mL per 12-h light period per photobioreactor and was observed at a growth density corresponding to 3.6 to 4.6 microgram Chl a. mL(-1) (1.2 to 1.6 mg dry weight. mL(-1)). Replacement of air with an argon atmosphere enhanced H(2) evolution by a factor of 2. This stimulatory effect was caused mainly by N(2) deprivation in the cell suspension. A short-term decrease of the CO(2) concentration in the air suppressed H(2) evolution. Anoxygenic conditions over the dark periods had a negative effect on H(2) production. The peculiarity of hydrogen production and some physiological characteristics of A. variabilis PK84 during cultivation in the photobioreactor under a light-dark regime are investigated.     

H2, N2, and O2 metabolism by isolated heterocysts from Anabaena sp. strain CA.

Metabolically active heterocysts isolated from wild-type Anabaena sp. strain CA showed high rates of light-dependent acetylene reduction and hydrogen evolution. These rates were similar to those previously reported in heterocysts isolated from the mutant Anabaena sp. strain CA-V possessing fragile vegetative cell walls. Hydrogen production was observed with isolated heterocysts. The ratio of C2H4 to H2 produced ranged from 0.9 to 1.2, and H2 production exhibited unique biphasic kinetics consisting of a 1 to 2-min burst of hydrogen evolution followed by a lower, steady-state rate of hydrogen production. This burst was found to be dependent upon the length of the dark period immediately preceding illumination and may be related to dark-to-light ATP transients. The presence of 100 nM NiCl2 in the growth medium exerted an effect on both acetylene reduction and hydrogen evolution in the isolated heterocysts from strain CA. H2-stimulated acetylene reduction was increased from 2.0 to 3.2 mumol of C2H4 per mg (dry weight) per h, and net hydrogen production was abolished. A phenotypic Hup- mutant (N9AR) of Anabaena sp. strain CA was isolated which did not respond to nickel. In isolated heterocysts from N9AR, ethylene production rates were the same under both 10% C2H2-90% Ar and 10% C2H2-90% H2 with or without added nickel, and net hydrogen evolution was not affected by the presence of 100 nM Ni2+. Isolated heterocysts from strain CA were shown to have a persistent oxygen uptake of 0.7 mumol of O2 per mg (dry weight) per h, 35% of the rate of whole filaments, at air saturating O2 levels, indicating that O2 impermeability is not a requirement for active heterocysts.     

Microalgae cultivation in air-lift reactors: modeling biomass yield and growth rate as a function of mixing frequency

The slow development of microalgal biotechnology stems from the failure in the design of large-scale photobioreactors where light energy is efficiently utilized. Due to the light gradient inside the reactor and depending on the mixing properties, algae are subjected to certain light/dark cycles where the light period is characterized by a light gradient. These light/dark cycles will determine productivity and biomass yield on light energy. Air-lift reactors can be used for microalgae cultivation and medium-frequency light/dark cycles will be found in these systems. Light/dark cycles are associated with two basic parameters: first, the light fraction, i.e., the ratio between the light period and the cycle time and second, the frequency of the light/dark cycle. In the present work, light/dark cycles found in air-lift reactors were simulated taking into account the light gradient during the light period. The effect of medium-frequency cycle time (10-100 s) and light fraction (0.1-1) on growth rate and biomass yield on light energy of the microalgae Dunaliella tertiolecta was studied. The biomass yield and growth rates were mainly affected by the light fraction, while cycle time had little influence. Response surface methodology was used and a statistical model describing the effect of light fraction and cycle time on growth rate and biomass yield on light energy was developed. The use of the model as a reactor design criterion is discussed.     

Sustained photoevolution of molecular hydrogen in a mutant of Synechocystis sp. strain PCC 6803 deficient in the type I NADPH-dehydrogenase complex

The interaction between hydrogen metabolism, respiration, and photosynthesis was studied in vivo in whole cells of Synechocystis sp. strain PCC 6803 by continuously monitoring the changes in gas concentrations (H2, CO2, and O2) with an online mass spectrometer. The in vivo activity of the bidirectional [NiFe]hydrogenase [H2:NAD(P) oxidoreductase], encoded by the hoxEFUYH genes, was also measured independently by the proton-deuterium (H-D) exchange reaction in the presence of D2. This technique allowed us to demonstrate that the hydrogenase was insensitive to light, was reversibly inactivated by O2, and could be quickly reactivated by NADH or NADPH (+H2). H2 was evolved by cells incubated anaerobically in the dark, after an adaptation period. This dark H2 evolution was enhanced by exogenously added glucose and resulted from the oxidation of NAD(P)H produced by fermentation reactions. Upon illumination, a short (less than 30-s) burst of H2 output was observed, followed by rapid H2 uptake and a concomitant decrease in CO2 concentration in the cyanobacterial cell suspension. Uptake of both H2 and CO2 was linked to photosynthetic electron transport in the thylakoids. In the ndhB mutant M55, which is defective in the type I NADPH-dehydrogenase complex (NDH-1) and produces only low amounts of O2 in the light, H2 uptake was negligible during dark-to-light transitions, allowing several minutes of continuous H2 production. A sustained rate of photoevolution of H2 corresponding to 6 micro mol of H2 mg of chlorophyll(-1) h(-1) or 2 ml of H2 liter(-1) h(-1) was observed over a longer time period in the presence of glucose and was slightly enhanced by the addition of the O2 scavenger glucose oxidase. By the use of the inhibitors DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea] and DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), it was shown that two pathways of electron supply for H2 production operate in M55, namely photolysis of water at the level of photosystem II and carbohydrate-mediated reduction of the plastoquinone pool.     

Conditioned medium factor produced and released by <Emphasis Type="Italic">Desmosdemus subspicatus</Emphasis> and its effect on the cell cycle of the producer

Conditioned medium (CM) containing algal exudates released by cells of Desmodesmus subspicatus (green microalga) enhanced proliferation of the producer in a dilution-dependent manner, and twofold diluted CM (CM/2) appeared to be the most effective. Asynchronous and synchronized by light/dark regime (14/10 h) cultures of D. subspicatus were used to explain the effects of CM/2 on photosynthesis as well as growth and reproductive processes of cell cycle of the producer. During the light period, all control cells attained three commitment points (CP) triggering the reproductive processes and 20% of them, additionally the fourth one. This resulted in the formation of mainly eight and partly 16 autospores released from the parent cell during the dark period of the cell cycle. CM/2 markedly increased the number of cells that attained the fourth CP. Chlorophyll a fluorescence parameters, measured by OJIP test (O, J, I, and P-different steps of fluorescence induction curve), were applied to explain stimulatory effect of CM/2 on the rate of oxygen evolution. The number of cells as well as the parameter of their “vitality” (P.I) determined by OJIP analysis were equally enhanced by CM/2. The maximum yield of electron trapping and transport, as well as the fraction of active reaction centers, was also significantly higher in CM/2. The cells quickly produced and released CM factor (CMF) to the culture medium; its highest activity was observed in the middle light phase of the cell cycle. Dialysis, ion exchange chromatography, and solid-phase extraction with C₁₈ allowed reproducible extraction of the active compound from CM. Subjecting the extract to reverse-phase high-performance liquid chromatography led to one active fraction eluted as a peak with the shortest retention time indicating its hydrophilicity.     

Relation between nitrogenase synthesis and activity in a marine unicellular diazotrophic strain, Gloeothece sp. 68DGA (Cyanophyte), grown under different light/dark regimens

The relationship between the abundance of nitrogenase and its activity was studied in the marine unicellular cyanobacterium Gloeothece sp. 68DGA cultured under different light/dark regimens. The Fe‐ and MoFe‐protein of nitrogenase and nitrogen (N₂)‐fixing (acetylene reduction) activity were detected only during the dark phase when the cells were grown under a 12 h light/12 h dark cycle (12L/12D). Nitrogenase activity appeared about 4 h after entering the dark phase. Maximum nitrogenase activity occurred at around the middle of the dark phase, and the activity rapidly decreased to zero before the start of the light phase. The rapid decrease of nitrogenase activity and the Fe‐protein of nitrogenase near the end of the dark phase in 12L/12D were partly recovered by the addition of l ‐methionine‐sulfoximine, an inhibitor of glutamine synthetase. Diurnal oscillation of the abundance of nitrogenase was maintained in the first subjective dark phase (i.e. the period corresponding to the dark phase) after the cells were transferred from 12L/12D to continuous illumination. However, enzyme activity was detected only when photosynthetic oxygen (O₂) evolution was completely suppressed by reducing the light intensity or by the addition of 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea. Nitrogenase always appeared in the cells about 16 h after starting the light phase, even when the 12L/12D cycle was modified by the addition or subtraction of a single 6 h period of light or dark. These results suggest the following: (i) N₂‐fixation by Gloeothece sp. 68DGA is primarily regulated by an endogenous circadian oscillator at the level of nitrogenase synthesis. (ii) The endogenous circadian rhythm resets on a shift of the timing of the light phase. (iii) Nitrogenase activity is not always reflected in the presence of nitrogenase. (iv) The activity of nitrogenase is negatively regulated by fixed nitrogen and the concentration of ambient O₂.     

大豆萌发过程的活性氧代谢

本文研究了大豆萌发过程中活性氧的产生与清除,并探讨了光因子在活性氧代谢中的作用。大豆呼吸强度、O产生速率及H2O2水平都在吸水后第四天达到高峰,然后下降,三者的变化趋势同步。SOD、POD及APX的活性随萌发过程而逐渐增强,最后趋于平稳。SOD同工酶谱中分别于萌发的第二、第三天各出现一条新的酶带。CAT在萌发的初期猛增50倍左右,之后趋于稳定。在三种清除H2O2的酶（CAT、POD、APX）中,CAT清除H2O2的能力远远高于POD与APX,CAT可能是大豆萌发过程中最主要的H2O2清除酶。光萌发时呼吸强度低于暗中萌发,但O产生速率与H2O2水平高于暗萌发,光萌发时O的产生占总耗氧量的1．1—2．7％,而暗中萌发为0．9—1．3％。光条件下SOD、APX活性明显高于暗中萌发,而POD与CAT则在光和暗条件下相差不大。     

星型孢菌素(STS)对蚕豆气孔运动的调控效应

用激光扫描共聚焦显微技术,初步研究广谱性蛋白激酶抑制剂星型孢菌素(STS)对蚕豆气孔运动的调控效应.结果表明:(1)光下STS对气孔开度无影响但暗中显著促进气孔开放,表明蛋白激酶参与光/暗对气孔运动的调控,光下蛋白激酶活性低而暗中高;(2)与H2O2清除剂抗坏血酸(ASA)和NO清除剂羧基-2-苯-4,4,5,5-四甲基咪唑-1-氧-3-氧化物(cPTIO)一样,STS既降低暗处理和光下外源H2O2、硝普钠(SNP)处理保卫细胞H2O2、NO水平,也促进气孔开放,表明暗中蛋白激酶通过抑制H2O2、NO清除机制提高保卫细胞内源H2O2、NO水平并促进气孔关闭.     

Role of carbonyl cyanide m-chlorophenylhydrazone in enhancing photobiological hydrogen production by marine green alga Platymonas subcordiformis

We demonstrated that a significant volume of H(2) gas could be photobiologically produced by a marine green alga Platymonas subcordiformis when an uncoupler of photophosphorylation, carbonyl cyanide m-chlorophenylhydrazone (CCCP), was added after 32 h of anaerobic dark incubation, whereas a negligible volume of H(2) gas was produced without CCCP. The role of CCCP in enhancing photobiological H(2) production was delineated. CCCP as an ADRY agent (agent accelerating the deactivation reactions of water-splitting enzyme system Y) rapidly inhibited the photosystem II (PSII) activity of P. subcordiformis cells, resulting in a markedly decline in the coupled oxygen evolution. The mitochondrial oxidative respiration was only slightly inactivated by CCCP, which depleted O(2) in the light. As a result, anaerobiosis during the stage of photobiological H(2) evolution was established, preventing severe O(2) inactivation of the reversible hydrogenase in P. subcordiformis. The uncoupling effect of CCCP accelerates electron transfer from water due to a disruption of the proton motive force and release of DeltapH across the thylakoid membrane and thus enhances the accessibility of electron and H(+) to hydrogenase. The electrons for hydrogen photoevolution are mainly from the photolysis of water (90%). Upon the addition of CCCP, Chl a/b ratio increased, which implies a decrease in the light-harvesting PSII antennae or an increase in PSII/PSI ratio, possibly resulting in higher efficiency of utilization of light energy. The enhancement of H(2) evolution by the addition of CCCP is mostly due to the combination of the above three mechanisms. However, the disruption of the proton gradient across the thylakoid membrane may prevent a sustained photobiological H(2) evolution due to a shortfall of ATP generation essential for the maintenance and repair functions of the cells.     

Aerobic hydrogen production by the heterocystous cyanobacteria Anabaena spp. strains CA and 1F.

Aerobic photoproduction of H2 was demonstrated in Anabaena spp. strains CA and 1F when cells were growing under nitrogen-fixing conditions. The rates of production, measured either by the hydrogen electrode or in a flow system by gas chromatography, were 10 to 15% of the rate of photosynthetic O2 evolution or 50 to 80% of the rates of acetylene reduction. Strains CA and 1F differed in several respects. In strain CA, H2 production was immediately partially sensitive to 3-(3,4-dichlorophenyl)-1,1-dimethylurea, whereas strain 1F was not immediately affected. Strain CA also showed a consistently higher rate of H2 production than did strain 1F. H2 production in strain CA was also markedly influenced by the light intensity used for growth, although the growth rates indicated that the light intensities used were essentially saturating.