Free radical mechanisms in enzyme reactions

Free radicals are formed in prosthetic groups or amino acid residues of certain enzymes. These free radicals are closely related to the activation process in enzyme catalysis, but their formation does not always result in the formation of substrate free radicals as a product of the enzyme reactions. The role of free radicals in enzyme catalysis is discussed.     

Hydrogen evolution by co-immobilized Chlorella vulgaris and Clostridium butyricum cells

Whole cells of Chlorella vulgaris and Clostridium butyricum were co-immobilized in 2% agar gel. NADP was suitable as an electron carrier. The rate of hydrogen evolution increased with increasing NADP concentration. The optimum conditions for hydrogen evolution were pH 7.0 and 37°C. The immobilized C. vulgaris-NADP-immobilized Cl. butyricum system continuously evolved hydrogen at a rate of 0.29–1.34 μmol/h per mg Chl for 6 days. On the other hand, the system without NADP evolved only a trace amount of hydrogen.     

Active site organization of bacterial type I fatty acid synthetase

Four kinds of active sites of bacterial fatty acid synthetase were mapped on distinct regions within a subunit. Active sites were specifically labeled with radioactive substrates and active-site-directed inhibitors. Labeled enzymes were cleaved with proteases, and the fragments thus produced were identified with respect to specific labels by SDS-polyacrylamide gel electrophoresis and a fluorographic technique. The linear alignment of such fragments in the original subunit was established and when the results were combined with those of our previous work, five active sites were located in three regions as follows. Starting from the N-terminal of the subunit, we located acetyl, malonyl and palmitoyl transferases in the first region, the acyl carrier site in the second region (Morishima & Ikai (1985) Biochim. Biophys. Acta 832, 297-307), and beta-ketoacyl synthetase in the third region. The observed order of active sites of bacterial fatty acid synthetase can be correlated with that of the yeast enzyme, which has two kinds of subunits.     

Study of the specific heme orientation in reconstituted hemoglobins

NMR studies of the recombination reaction of apohemoglobin derivatives with natural and unnatural hemes and of the heme-exchange reaction for reconstituted hemoglobin have revealed that the heme is incorporated into the apoprotein with stereospecific heme orientations dependent upon the heme peripheral 2,4-substituents and the axial iron ligand(s). Heme orientations also depend on whether recombination occurs at the alpha or beta subunit and on whether or not the complementary subunit is occupied by the heme. In the recombination reaction with the azido complex of deuterohemin, the alpha subunit of the apohemoglobin preferentially combines with the hemin in the "disordered" heme orientation, whereas protohemin is inserted in either of two heme orientations. Mesohemin inserts predominantly in the "native" heme orientation. For the beta subunit, specific heme orientation was also encountered, but the specificity was somewhat different from that of the alpha subunit. It was also shown that the specific heme orientation in both subunits is substantially affected by the axial heme ligands. These findings imply that apohemoglobin senses the steric bulkiness of both the porphyrin 2,4-substituents and the axial iron ligands in the heme-apoprotein recombination reaction. To gain an insight into the effect of the protein structure, the heme reconstitution reaction of semihemoglobin, demonstrating that the heme orientation in the reconstituted semihemoglobin with the azido-deuterohemin complex was in the native form, was also examined.(ABSTRACT TRUNCATED AT 250 WORDS)     

Vegetative growth and productivity ofEichhornia azurea with special emphasis on leaf dynamics

The lifeform and the biological production of pure stands ofEichhornia azurea Kunth in three lakes in tropical Brazil were studied. The lifeform ofE. azurea is termed ‘semi-emergent’, because the plant has well developed trailing stems just under the water, and the aerial lamina emerges with the thick petiole. The density of shoot apices was 9.9, 17.2 and 17.1 m⁻² in Lake Dom Helvecio, Lake Jacaré and Lake Carioca, respectively. The mean daily increment of the apical shoot biomass was between 1.8 and 4.8 g m⁻² day⁻¹. The mean leaf life-span in Lake Dom Helvecio, Lake Jacaré and Lake Carioca was estimated to be 78, 49 and 64 days in the wet season and 73, 70 and 73 days in the dry season, respectively. The stem life-span was estimated to be about 28 months. Starch content in the current years' stem ranged from 24 to 118 mg g⁻¹ dry matter with fluctuations, the amplitude of which decreased with age. The differences for most of the growth parameters, such as density of shoot apices, daily increment of biomass and leaf life-span, between dry and wet season are smaller than those among the three lakes. Both the decrease in daily dry matter production and the increase in leaf life-span occurred in order from Lake Dom Helvecio to Lake Jacaré and Lake Carioca. The low productivity ofE. azurea is considered to be related to a low leaf area index, a long time interval for the emergence of new leaves, long leaf life-span and a low capacity for branching.     

A specific microbial sensor for formic acid

Summary A microbial sensor consisting of immobilized Clostridium butyricum, two gas permeable Teflon membranes and fuel cell type electrode was suitable for the determination of formic acid. When the sensor was inserted into the sample solution containing formic acid, the current increases to a steady state with a response time of 20 min. The relationship between the steady state current and the formic acid concentration is linear up to 1 000 mg l^–1. The currents are reproducible with an average relative error of 5%. Selectivity of the sensor is satisfactory. Results obtained with this sensor and by gas chromatography were in good agreement (regression coefficient; 0.98) when the cultivation medium of Aeromonas formicans was employed. Immobilized Clostridium butyricum is stable for more than 20 days.     

Occurrence and induction of spermidine-N1-acetyltransferase in Escherichia coli

Spermidine acetylase activity was detected in extracts prepared from $\text{Escherichia coli}$ and there was a marked increase in activity over the early period of growth. This increase reached a maximum 3 h after inoculation and was followed by an increase in ornithine decarboxylase activity. The acetylase was also able to use spermine as a substrate, but not putrescine. With spermidine and acetyl-CoA as substrate, the product formed was exclusively N¹-acetyl-spermidine. This is the first evidence for the occurrence in bacteria of spermidine-N¹-acetyltransferase, an enzyme which has previously been described in mammalian cells. These results suggest that acetylation of spermidine may be involved in the growth of $\text{Escherichia coli}$ and in the regulation of its polyamine content.