Influence of increased dissolved oxygen concentration on productivity and selectivity in cultures of a colabomycin-producing strain ofStreptomyces griseoflavus

The influence of enhanced O₂ concentration on growth and formation of secondary metabolites byStreptomyces griseoflavus (strain Tü 2880) was investigated in a stirred tank and in an air-lift fermentor. At a partial pressure of O₂ p_o2 = 1880 mbar the growth was lowered by 50% compared to p_o2 = 210 mbar, whilst substrate consumption and O₂ uptake rate increased markedly. Production of the colabomycin complex reached maximum values at p_o2 = 630 mbar. A similar increase of secondary metabolite formation was obtained when glycerol or acetate were fed at p_o2 = 220 mbar. The portion of the derivate colabomycin A in the product mixture rose from 43% at p_o2 = 210 mbar to 73% at p_o2 = 1260 mbar. Since dissolved O₂ concentration has a significant influence on productivity and selectivity it may be used to regulate aerobic fermentation processes.     

Insulin Sensitivity,Cardiorespiratory Fitness,and Physical Activity in Overweight Hispanic Youth

Objective: To determine whether cardiorespiratory fitness and/or physical activity (PA) were related to measures of insulin sensitivity and secretion independent of body composition in overweight Hispanic children. Research Methods and Procedures: Ninety‐five Hispanic children (n = 55 boys; n = 40 girls; 8 to 13 years old) participated in this investigation. The frequently sampled intravenous glucose tolerance test was used to determine the insulin sensitivity index (SI), the acute insulin response, and the disposition index. Cardiorespiratory fitness [maximal oxygen uptake (Vo_2max)] was evaluated using a treadmill protocol, and PA was determined by an interviewer‐administered questionnaire. Body composition was measured using DXA. Results: Unadjusted correlations indicated that Vo_2max (milliliters of O₂ per minute) was negatively related to SI (r = ?0.46, p < 0.05) and disposition index (r = ?0.31, p < 0.05) and positively associated with fasting insulin (r = 0.29, p < 0.05), but these relationships were no longer significant once gender, Tanner stage, fat mass, and soft lean tissue mass were included as covariates (all p > 0.05). Multivariate linear regression analysis showed that body fat mass explained 53% of the variance in SI and that Vo_2max (milliliters of O₂ per minute) was not independently related to SI. Cardiorespiratory fitness was positively related to both fat mass (r = 0.43, p < 0.001) and soft lean tissue mass (r = 0.89, p < 0.001). PA was not related to any measure of insulin sensitivity and secretion. Discussion: Cardiorespiratory fitness, as determined by Vo_2max (milliliters of O₂ per minute), was not independently related to insulin sensitivity or secretion, suggesting that Vo_2max influences insulin dynamics indirectly through fat mass.     

Screening of Bacterial Isolates from Polluted Soils Exhibiting Catalase and Peroxidase Activity and Diversity of their Responses to Oxidative Stress

For the survival of individual isolates of gram-negative bacteria Pseudomonas putida, Achromobacter xylosoxidans, and the gram-positive bacterium Bacillus megaterium, in an environment polluted with crude oil products, the production of catalases exhibiting both catalase and dianisidine-peroxidase activity is important. Electrophoretic resolution of cell-free extracts of aerobically grown strains in Luria–Bertani medium during exponential phase revealed distinctive expression of catalatic and peroxidatic activities detected with 3,3′-diaminobenzidine tetrahydrochloride. A considerable diversity in microbial catalase and peroxidase responses to 20 or 40 mM H₂O₂ stress, resulted from hydroperoxidase’s variant of original isolates, indicating an environmental selective pressure. However, catalase was important for the adaptation of cultures to high concentration of 60 mM H₂O₂. Appreciable differences in the sensitivity to toxic effect of H₂O₂ (20 or 40 mM) treatment between individual isolates and their adapted variants during growth were observed until the middle of exponential phase, but they were insignificant at the entry to stationary phase. Isolates also exhibited a considerable diversity in catalases responses to phenolic contaminants 1 and 2 mM o- or p-phenylenediamine. Catalase activity of bacterium P. putida was visibly stimulated only by p-phenylenediamine and not by its positional isomer o-PDA. This study contributes to a better understanding of the role catalases play in bacterial responses to a polluted environment.     

Phenoloxidases from larval cuticle of the sheep blowfly,Lucilia cuprina: Characterization,developmental changes,and inhibition by antiphenoloxidase antibodies

A tyrosinase, enzyme A (EC 1.10.3.1, o-diphenol: O₂ oxidoreductase), and a laccase, enzyme B (EC 1.10.3.2, p-diphenol: O₂ oxidoreductase), have been partially purified and characterized from larval cuticle of the sheep blowfly, Lucilia cuprina. Enzyme A is active toward a range of o-diphenols but not p-diphenols, is strongly inhibited by thiourea and phenylthiourea, has a pH optimum between 6.5 and 7.0, and yields a single, 60,000 molecular weight subunit following SDS gel electrophoresis. Enzyme B is active toward both o-diphenols and p-diphenols, is only slightly inhibited by phenylthiourea, has a pH optimum near 4.5, is highly thermostable, and has an apparent molecular weight of 90,000. Enzyme A appears to be activated from an inactive proenzyme in the cuticle and to be present throughout the wandering phase of the final larval instar, declining at pupariation. Enzyme B is present in active form, increases greatly in the cuticle just at the time of pupariation, and then decreases as sclerotization occurs. Antibodies against enzyme A have been raised in sheep and rabbits, and against enzyme B in rabbits, but diets containing antiphenoloxidase antibodies did not affect development or mortality of fly larvae.     

Isolation of the Bromo Analogues of Pyrrolnitrin from Pseudomonas pyrrolnitrica

The metabolism of an organophosphorous fungicide, Kitazin P® (S-benzyl O, O-diisopropyl phosphorothioate) by mycelia of Pyricularia oryzae, the rice blast fungus, was studied by using ³²P- and ³⁵S-doubled-labeled, ³⁵S-labeled and non-labeled compounds.

The mycelia incorporated the fungicide from the medium, metabolized most of it to water-soluble metabolites and released them into the medium. The main metabolite from Kitazin P was identefied as O, O-diisopropyl hydrogen phosphorothioate by ion-exchange chromatographic separation of double-labeled metabolites. S-Benzyl O-isopropyl hydrogen phosphorothioate, diisopropyl hydrogen phosphate and/or isopropyl dihydrogen phosphate were suspected to be minor metabolites. From the toluene-soluble fraction several metabolites were separated by thin-layer chromatography. The most eminent metabolite among them was found to be the m-hydroxy derivative of Kitazin P by thin-layer chromatographic identification of the labeled metabolite and gas-liquid chromatographic identification of the metabolite and its methylated and acetylated products with respective authentic samples. The o- and p-hydroxy derivatives were not found in the metabolites.

No significant difference in the rate and extent of metabolism was found between susceptible and resistant clones against the fungicide, when its concentration was below the growth-inhibitory concentration to the both clones.     

Regulatory O2 tensions for the synthesis of fermentation products in Escherichia coli and relation to aerobic respiration

In an oxystat, the synthesis of the fermentation products formate, acetate, ethanol, lactate, and succinate of Escherichia coli was studied as a function of the O₂ tension (pO₂) in the medium. The pO₂ values that gave rise to half-maximal synthesis of the products (pO_0.5) were 0.2–0.4 mbar for ethanol, acetate, and succinate, and 1 mbar for formate. The pO_0.5 for the expression of the adhE gene encoding alcohol dehydrogenase was approximately 0.8 mbar. Thus, the pO₂ for the onset of fermentation was distinctly lower than that for anaerobic respiration (pO_0.5≤ 5 mbar), which was determined earlier. An essential role for quinol oxidase bd in microaerobic growth was demonstrated. A mutant deficient for quinol oxidase bd produced lactate as a fermentation product during growth at microoxic conditions (approximately 10 mbar O₂), in contrast to the wild-type or a quinol-oxidase-bo-deficient strain. In the presence of nitrate, the amount of lactate was largely decreased. Therefore, under microoxic conditions, the pO₂ appears to be too high for (mixed acid) fermentation to function and too low for aerobic respiration by quinol oxidase bo. Received: 7 February 1997 / Accepted: 2 May 1997     

Analysis of the metabolites of the sodium salt of 6-hydroxy-5-(phenylazo)-2-naphthalenesulfonic acid in Sprague–Dawley rat urine

The sodium salt of 6-hydroxy-5-(phenylazo)-2-naphthalenesulfonic acid (SS-AN), which is a subsidiary color present in Food Yellow No. 5 [Sunset Yellow FCF, disodium salt of 6-hydroxy-5-(4-sulfophenylazo)-2-naphthalenesulfonic acid], was orally administered to Sprague–Dawley rats. Metabolite A, metabolite B, and unaltered SS-AN were detected as colored metabolites in the rat urine. Analysis of the chemical structures showed that metabolite A (major peak) was 6-hydroxy-5-(4-sulfooxyphenylazo)-2-naphthalenesulfonic acid, the sulfuric acid conjugate of SS-AN, and metabolite B (minor peak) was 6-hydroxy-5-(4-hydroxyphenylazo)-2-naphthalenesulfonic acid (SS-PAP), which is a derivative of metabolite A without the sulfuric acid. The colorless metabolites p-aminophenol, o-aminophenol, and aniline present in the urine were analyzed by liquid chromatography–mass spectrometry. The orally administered SS-AN had been metabolized to the colorless metabolites (p-aminophenol 45.3%, o-aminophenol 9.4%, aniline 0.4%) in the 24-h urine samples. Analysis of the colored metabolites by high-performance liquid chromatography with detection at 482 nm indicated the presence of metabolite A (0.29%), SS-PAP (0.01%), and SS-AN (0.02%) were detected in the 24-h urine samples. Approximately 56% of SS-AN was excreted into the urine and the rest is probably excreted into feces.     

Photochemistry of the O-Nitrobenzyl Protecting Group in RNA Synthesis

Abstract

UV irradiation of 2′-O (o-nitrobenzyl)adenylyl(3′-5′)uridine in the presence of O₂ yields the corresponding nitrobenzoyl derivative in addition to the expected A-U. A mechanism proposed for this oxidation involves the successive removal of the two benzylic protons with a hydroperoxide as the intermediate between the two steps.     

Availability of O2 as a Substrate in the Cytoplasm of Bacteria under Aerobic and Microaerobic Conditions

The growth rates of Pseudomonas putida KT2442 and mt-2 on benzoate, 4-hydroxybenzoate, or 4-methylbenzoate showed an exponential decrease with decreasing oxygen tensions (partial O₂ tension [pO₂] values). The oxygen tensions resulting in half-maximal growth rates were in the range of 7 to 8 mbar of O₂ (corresponding to 7 to 8 μM O₂) (1 bar = 10⁵ Pa) for aromatic compounds, compared to 1 to 2 mbar for nonaromatic compounds like glucose or succinate. The decrease in the growth rates coincided with excretion of catechol or protocatechuate, suggesting that the activity of the corresponding oxygenases became limiting. The experiments directly establish that under aerobic and microaerobic conditions (about 10 mbar of O₂), the diffusion of O₂ into the cytoplasm occurs at high rates sufficient for catabolic processes. This is in agreement with calculated O₂ diffusion rates. Below 10 mbar of O₂, oxygen became limiting for the oxygenases, probably due to their high K_m values, but the diffusion of O₂ into the cytoplasm presumably should be sufficiently rapid to maintain ambient oxygen concentrations at oxygen tensions as low as 1 mbar of O₂. The consequences of this finding for the availability of O₂ as a substrate or as a regulatory signal in the cytoplasm of bacterial cells are discussed.     

Using combined measurements of gas exchange and chlorophyll fluorescence to estimate parameters of a biochemical C3 photosynthesis model: a critical appraisal and a new integrated approach applied to leaves in a wheat (Triticum aestivum) canopy

We appraised the literature and described an approach to estimate the parameters of the Farquhar, von Caemmerer and Berry model using measured CO₂ assimilation rate (A) and photosystem II (PSII) electron transport efficiency (Φ₂). The approach uses curve fitting to data of A and Φ₂ at various levels of incident irradiance (I_inc), intercellular CO₂ (C_i) and O₂. Estimated parameters include day respiration (R_d), conversion efficiency of I_inc into linear electron transport of PSII under limiting light [κ_2(LL)], electron transport capacity (J_max), curvature factor (θ) for the non‐rectangular hyperbolic response of electron flux to I_inc, ribulose 1·5‐bisphosphate carboxylase/oxygenase (Rubisco) CO₂/O₂ specificity (S_c/o), Rubisco carboxylation capacity (V_cmax), rate of triose phosphate utilization (T_p) and mesophyll conductance (g_m). The method is used to analyse combined gas exchange and chlorophyll fluorescence measurements on leaves of various ages and positions in wheat plants grown at two nitrogen levels. Estimated S_c/o (25 °C) was 3.13 mbar µbar^?1; R_d was lower than respiration in the dark; J_max was lower and θ was higher at 2% than at 21% O₂; κ_2(LL), V_cmax, J_max and T_p correlated to leaf nitrogen content; and g_m decreased with increasing C_i and with decreasing I_inc. Based on the parameter estimates, we surmised that there was some alternative electron transport.     

Synthesis and structure–activity relationship of pyripyropene A derivatives as potent and selective acyl-CoA:cholesterol acyltransferase 2 (ACAT2) inhibitors: Part 3

In an effort to develop potent and selective inhibitors toward ACAT2, structure–activity relationship studies were carried out using derivatives based on pyripyropene A (PPPA, 1). In particular, we investigated the possibility of introducing appropriate 1,11-O-benzylidene and 7-O-substituted benzoyl moieties into PPPA (1). The new o-substituted benzylidene derivatives showed higher selectivity for ACAT2 than PPPA (1). Among them, 1,11-O-o-methylbenzylidene-7-O-p-cyanobenzoyl PPPA derivative 7q and 1,11-O-o,o-dimethylbenzylidene-7-O-p-cyanobenzoyl PPPA derivative 7z proved to be potent ACAT2 inhibitors with unprecedented high isozyme selectivity.     

The oxygen supply to thermogenic flowers

Thermogenic flowers produce heat by intense respiration, and the rates of O₂ consumption (?o₂) in some species can exceed those of all other tissues of plants and most animals. By exposing intact flowers to a range of O₂ pressures (Po₂) and measuring ?o₂, we demonstrate that the highest respiration rates exceed the capacity of the O₂ diffusive pathway and become diffusion limited in atmospheric air. The male florets on the inflorescence of Arum concinnatum have the highest known mass‐specific ?o₂ and can be severely diffusion limited. Intact spadices of Japanese skunk cabbage Symplocarpus renifolius are diffusion limited in air only when ?o₂ is maximal, but not at lower levels. True flowers of the sacred lotus Nelumbo nucifera and the appendix of Arum concinnatum are never diffusion limited in air. ?o₂ – Po₂ curves are evaluated quantitatively with the ‘Regulation Index’, a new tool to measure dependence of ?o₂ on ambient Po₂, as well as the conventional ‘Critical Po₂’. The study also includes measurements of Po₂ within thermogenic tissues with O₂‐sensitive fibre optics, and reveals that the diffusion pathway is complicated and that O₂ can be provided not only from the surface of the tissues but also from the pith of the flower's peduncle.     

Effect of Photon Fluence Rate on Oxygen Evolution and Uptake by Chlamydomonas reinhardtii Suspensions Grown in Ambient and CO(2)-Enriched Air

A closed system consisting of an assimilation chamber furnished with a membrane inlet from the liquid phase connected to a mass spectrometer was used to measure O₂ evolution and uptake by Chlamydomonas reinhardtii cells grown in ambient (0.034% CO₂) or CO₂-enriched (5% CO₂) air. At pH = 6.9, 28°C and concentrations of dissolved inorganic carbon (DIC) saturating for photosynthesis, O₂ uptake in the light (U_o) equaled O₂ production (E_o) at the light compensation point (15 micromoles photons per square meter per second). E_o and U_o increased with increasing photon fluence rate (PFR) but were not rate saturated at 600 micromoles photons per square meter per second, while net O₂ exchange reached a saturation level near 500 micromoles photons per square meter per second which was nearly the same for both, CO₂-grown and air-grown cells. Comparison of the U_o/E_o ratios between air-grown and CO₂-grown C. reinhardtii showed higher values for air-grown cells at light intensities higher than light compensation. For both, air-grown and CO₂-grown algae the rates of mitochondrial O₂ uptake in the dark measured immediately before and 5 minutes after illumination were much lower than U_o at PFR saturating for net photosynthesis. We conclude that noncyclic electron flow from water to NADP⁺ and pseudocyclic electron flow via photosystem I to O₂ both significantly contribute to O₂ exchange in the light. In contrast, mitochondrial respiration and photosynthetic carbon oxidation cycle are regarded as minor O₂ consuming reactions in the light in both, air-grown and CO₂-grown cells. It is suggested that the “extra” O₂ uptake by air-grown algae provides ATP required for the energy dependent CO₂/HCO₃⁻ concentrating mechanism known to be present in these cells.     

Evaluation of a Multi-parameter Biomarker Set for Oxidative Damage in Man: Increased Urinary Excretion of Lipid,Protein and DNA Oxidation Products after One Hour of Exercise

The objective of the present study was to evaluate a comprehensive set of urinary biomarkers for oxidative damage to lipids, proteins and DNA, in man. Eighteen moderately trained males (mean age 24.6±0.7) exercised 60?min at 70% of maximal O₂ uptake on a cycle ergometer. Urine fractions for 12?h were collected 1 day before, and for 3 consecutive days after exercise.

As biomarkers of lipid peroxidation, 8 aldehydes (i.e. propanal, butanal, pentanal, hexanal, heptanal, octanal, nonanal and malondialdehyde—MDA)and acetone were analyzed in urines by gas chromatography with electron capture detection (GC-ECD). As a biomarker of protein oxidation, o,o′-dityrosine was analyzed in urine samples by a recently developed isotope dilution HPLC-atmospheric pressure chemical ionization (APCI)-tandem-mass spectrometry (HPLC-APCI-MS/MS) methodology. As a biomarker of oxidative DNA damage, urinary excretion of 8-hydroxy-2′-deoxyguanosine (8-OHdG) was measured by an ELISA method.

On the day of exercise, significant increases were observed in urinary excretions of acetone (?p<0.025, n=18) and butanal (?p<0.01, n=18) in the 12?h daytime fractions compared to the daytime fraction before exercise. The urinary acetone excretion was also significantly (?p<0.05) increased on the 1st day after exercise. Octanal and nonanal were increased in the daytime urine fraction on the 2nd day after exercise. However, these increases were of borderline significance (?p=0.09 and p=0.07, respectively).

Significantly elevated urinary o,o′-dityrosine amounts were observed in the daytime fraction on the day of exercise (?p<0.025) and on the 1st day after exercise (?p=0.07) compared to the before exercise daytime fraction.

Excretion of urinary 8-OHdG was statistically significantly increased in the daytime fractions on the day of exercise (?p=0.07) and on the 1st day after exercise (?p<0.025) compared to before exercise daytime fraction.

Increases in urinary excretions of acetone, propanal, pentanal, MDA and 8-OHdG significantly correlated with training status (hours of exercise/week) of the volunteers, while o,o′-dityrosine did not.

To our knowledge, the present study is the first to evaluate a multi-parameter non-invasive biomarker set for damage to three main cellular targets of ROS. It shows that 1?h of exercise may already induce oxidative damage in moderately trained individuals and that the chosen urinary biomarkers are sensitive enough to monitor such damage.     

Oxygen uptake of carp,Cyprinus carpio,swimming in normoxic and hypoxic water

Synopsis Oxygen uptake (V_{o 2}) was measured in carp of approximately 40 cm length swimming at controlled variable oxygen tensions (P_{o 2}). At P_{o 2}> 120 mm Hg V_{o 2} increased with an increase in swimming speed from 5.6 to 11.3 cm · sec^–1. Extrapolation of V_{o 2} to zero activity at P_{o 2} = 140 mm Hg revealed a standard O₂ uptake of 36.7 ml O₂ · kg^–1 · h^–1 at 20° C. At the lowest swimming speed (5.6 cm · s^–1) the oxygen uptake increased when the water P_{o 2} was reduced. A near doubling in V_{o 2} was seen at P_{o 2} = 70 mm Hg compared to 140 mm Hg. At higher swimming speeds in hypoxic water V_{o 2} decreased relative to the values at low swimming speeds. As a result the slope of the lines expressing log V_{o 2} as a function of swimming speed decreased from positive to negative values with decreasing P_{o 2} of the water. pH of blood from the caudal vein drawn before and at termination of swimming at P_{o 2} = 70 mm Hg and 100 mm Hg did not show any decrease in relation to rest values at P_{o 2} = 140 mm Hg. Blood lactate concentration did not increase during swimming at these tensions.     

Partial Purification and Characterization of an Elastase-inhibitory Lipid Derivative from a Cyanobacterium, <Emphasis Type="Italic">Microcystis</Emphasis> Ku2

A unicyanobacterial isolate of cyanobacterium, identified as Microcystis Ku2, produced a mammalian elastase-inhibitory lipid derivative. Protease inhibitors in cyanobacteria are unequivocally peptides. Since this metabolite appeared in lipid phase, we worked on a hypothesis that whether metabolite other than peptides could be responsible for the characteristic inhibition. It was purified by saponification and reverse phase column chromatography. The resulting compound was tentatively characterized as a glycolipid with structure of sulfated di-pentose derivative of ceramide (MW = 956 Da). The apparent IC₅₀ for elastase was 1.3 μM.     

Simultaneous determination of Rubisco carboxylase and oxygenase kinetic parameters in Triticum aestivum and Zea mays using membrane inlet mass spectrometry

The lack of complete Rubisco kinetic data for numerous species is partly because of the time consuming nature of the multiple methods needed to assay all of the Rubisco parameters. We have developed a membrane inlet mass spectrometer method that simultaneously determines the rate of Rubisco carboxylation (v_c) and oxygenation (v_o), and the CO₂ and O₂ concentrations. Using the collected data, the Michaels‐Menten equations for v_c and v_o in response to changing CO₂ and O₂ concentrations were simultaneously solved for the CO₂ (K_c) and O₂ (K_o) constants, the maximum turnover rates of the enzyme for CO₂ (kcat_CO2) and O₂ (kcat_O2) and the specificity for CO₂ relative to O₂ (S_c/o). In the C₄ species Zea mays K_c was higher but K_o was lower compared with the C₃ species Triticum aestivum. The kcat_CO2 was higher and the kcat_O2 lower in Z. mays compared with T. aestivum and S_c/o was similar in the two species. The V_omax/V_cmax was lower in Z. mays and thus did not correlate with changes in S_c/o. In conclusion, this mass spectrometer system provides a means of simultaneously determining the important Rubisco kinetic parameters, K_c, K_o, kcat_CO2,kcat_O2 and S_c/o from the same set of assays.     

Releases of ingested phytoplankton carbon by Daphnia magna

1. The carbon budgets and assimilation efficiencies (AEs) of adults and juveniles of Daphnia magna were quantified using ¹⁴C as a tracer. Animals were fed pure Chlamydomonas reinhardtii or Scenedesmus obliquus at different food concentrations. Carbon AEs (46–70%) were comparable at food concentrations of 0.03–0.30 mg C L^?1 for both algal species, but decreased to 34–49% when the food concentration further increased by 10‐fold. The carbon AEs were significantly and negatively correlated with the food level. 2. During the postdigestive period, partitioning of ingested carbon into different compartments including excretion, respiration and egestion was not influenced by the food species and life stage. There was a negative correlation between respiration (as % of total loss) and food concentration and a positive correlation between egestion (as % of total loss) and food concentration. Dissolved organic carbon (DOC) and CO₂ accounted for 55–72% and 9–37%, respectively, of the total carbon loss from juveniles fed both algal diets. For adults, DOC and CO₂ contributed to 44–64% and 20–47% of the total carbon loss, respectively. Particulate organic carbon (POC) was a minor pathway for the overall carbon loss. 3. The turnover and release budget of structural carbon (as moults and neonate reproduction) were further evaluated in long‐term experiments at different algal concentrations. Food concentration did not affect the carbon efflux or the carbon allocation into different physiological compartments except for respiration. Juveniles had twofold lower carbon turnover rate (0.12–0.16 day^?1) than those of the adults (0.32–0.35 day^?1). In adults, comparable carbon was allocated into DOC (35–42%) and reproduction (27–35%), which were the dominant routes for carbon loss. For the juveniles, DOC accounted for 42–64% of the total carbon loss. 4. About 21–38% of the total DOC released by adults and juveniles was associated with the high molecular weight organic carbon fraction (>5 kDa). Our results show that carbon was mainly lost by D. magna in the form of DOC during assimilation process as well as from the structural materials. Reproduction or maternal transfer was another major drain of body carbon for adult D. magna.     

The chloro-o,o′-dinitrophenyl group: Properties and use in protein chemistry

The properties of the chloro-o,o′-dinitrophenyl group, more suitable as a building block for the design of chromogenic bifunctional protein reagents than the corresponding known o,p derivatives, are investigated by model reactions with small molecules and with proteins. This study shows that (1) the chlorine substituent can be easily substituted by protein nucleophiles, (2) arylation of SH and NH₂ groups leads to chromophores in which the differences between the λ_max (330 and 430 nm, respectively) are larger than in the case of o,p-dinitro compounds, (3) afler substitution by an SH group, transarylation to a vicinal NH₂, whenever possible, occurs faster than in the case of o,p derivatives (4) substitution by imidazole or phenol can be reversed by a thiol yielding an alkylthioaryl derivative; however, when mercaptoethanol is used in this reaction, o,o′-dinitrophenol is obtained, probably resulting from a S to O transfer in the initially formed hydroxyethylthiophenol, followed by hydrolysis of the labile alkoxyphenyl derivative.     

A biochemical model of photosynthetic CO2 assimilation in leaves of C3 species

Various aspects of the biochemistry of photosynthetic carbon assimilation in C₃ plants are integrated into a form compatible with studies of gas exchange in leaves. These aspects include the kinetic properties of ribulose bisphosphate carboxylase-oxygenase; the requirements of the photosynthetic carbon reduction and photorespiratory carbon oxidation cycles for reduced pyridine nucleotides; the dependence of electron transport on photon flux and the presence of a temperature dependent upper limit to electron transport. The measurements of gas exchange with which the model outputs may be compared include those of the temperature and partial pressure of CO₂(p(CO₂)) dependencies of quantum yield, the variation of compensation point with temperature and partial pressure of O₂(p(O₂)), the dependence of net CO₂ assimilation rate on p(CO₂) and irradiance, and the influence of p(CO₂) and irradiance on the temperature dependence of assimilation rate.Abbreviations RuP₂ ribulose bisphosphate - PGA 3-phosphoglycerate - C=p(CO₂) partial pressure of CO₂ - O=p(O₂) partial pressure of O₂ - PCR photosynthetic carbon reduction - PCO photorespiratory carbon oxidation