• Volume 4, Issue 1

      March 1982,   pages  1-124

    • Regulation of nitrate reduction in wheat leaves

      M S Naik D J D Nicholas

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      Wheat leaves exposed to 710 nm monochromatic light, when only photosystem 1 operates, reduced small but significant amount of nitrate to nitrite. This could be due to partial inhibition of mitochondrial oxidation of NADH, brought about by cyclic photo-phosphorylation. Under dark aerobic conditions, citric acid cycle intermediates only slightly stimulated nitrate reduction. Under dark anaerobic conditions, when maximum reduction of nitrate occurred, the time course showed a 1:1 stoichiometry between nitrite and CO2. It is suggested that for maximum reduction of nitrate under physiological conditions, CO2 fixation and export of ATP via triose phosphate shuttle is essential.

    • Prostaglandin synthetase activity of goat vesicular microsomes: Co-factor requirement and effect of calcium ions

      Dolly Gosh Joytirmoy Dutta

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      The effects of several co-factors and bivalent cations on the activity of prostaglandin synthetase isolated from goat seminal vesicles were studied. Ca2+ appears to play a regulatory role in the biosynthesis of prostaglandin E2 by goat vesicular microsomes as the normal parabolic time course of synthesis changed to a sigmoid curve in the presence of 4 mM Ca2+ and to nearly a hyperbolic pattern when the microsomes were preincubated with the metal ions. The Ca2+ modulated reaction showed increased rate of prostaglandin E2 synthesis only when the period of incubation was extended beyond 30 min. The co-factor requirement of the goat enzyme was similar to that of the bovine and ovine prostaglandin synthetase systems.

    • Changes in the content of glycogen and its metabolites during acute exposure ofAnabas testudineus (Bloch) to furadan

      R Bakthavathsalam Y Srinivasa Reddy

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      Significant differences were observed in glycogen metabolism ofAnabas testudineus exposed to an acute lethal (1.56 mg/litre) and a sublethal (0.56 mg/litre) concentration of furadan. At sublethal concentration, the muscle glycogen which was utilized during the early periods of exposure, was replenished in the later period of exposure and at 120 h, the muscle glycogen levels were higher than the control. At higher concentration, the liver glycogen levels showed an increase presumably at the expense of fuel reserves of the muscle.

    • Bacitracin A: Structure-activity relationship

      S Rambhav

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      The single imidazole nucleus of L-histidine residue in bacitracin-A seems to be important for the anti-bacterial activity of the molecule, since iodination, carboxymethylation and coupling of diazobenzene sulphonic acid to the histidine residue in the antibiotic caused 90–94% loss of antibacterial activity of the antibiotic. In contrast, the bacitracin sulphone and sulphoxide derivatives are as active as the parent antibiotic.

    • Purification, physicochemical and regulatory properties of serine hydroxymethyltransferase from sheep liver

      R Manohar K S Ramesh N Appaji Rao

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      Serine hydroxymethyltransferase (EC 2.1.2.1) was purified from the cytosolic fraction of sheep liver by ammonium sulphate fractionation, CM-Sephadex chromatography, gel filtration using Ultrogel ACA 34 and Blue Sepharose affinity chromatography. The homogeneity of the enzyme was rigorously established by Polyacrylamide gel and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, isoelectrofocusing, ultracentrifugation, immunodiffusion and Immunoelectrophoresis. The enzyme was a homotetramer with a molecular weight of 210,000 ±5000. The enzyme showed homotropic cooperative interactions with tetrahydrofolate (nH =2.8) and a hyperbolic saturation pattern with L-serine. At the lowest concentration of tetrahydrofolate used (0.2 mM), only 5% of the added folate was oxidized during preincubation and assay. ThenH value was independent of the time of preincubation. Preincubation of the enzyme with serine resulted in a partial loss of the cooperative interactions (nH =1.6) with tetrahydrofolate. The enzyme was regulated allosterically by interaction with nicotinamide nucleotides; NADH was a positive effector while NAD+ was a negative allosteric effector. The subunit interactions were retained even at the temperature optimum of 60‡C unlike in the case of the monkey liver enzyme, where these interactions were absent at higher temperatures. D-Cycloserine, a structural analogue of serine caused a sigmoid pattern of inhibition, in contrast with the observations on the monkey liver enzyme. Cibacron blue F3GA completely inhibited the enzyme and this inhibition could be reversed by tetrahydrofolate. Unlike in the monkey liver enzyme, NAD+ and NADH gave considerable protection against this inhibition. The sheep liver enzyme differs significantly in its kinetic and regulatory properties from the serine hydroxymethyltransferases isolated from other sources.

    • Immobilization of trypsin on sand: Mode of binding

      R Puvanakrishnan S M Bose B R Reddi

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      Acid-washed and heat-treated river sand was separated into different fractions by geochemical methods and immobilization of trypsin was carried out on the separated fractions using 3-aminopropyltriethoxysilane and glutaraldehyde. Scanning Electron Micrographs of the purified fraction (Sp. gr >2.5 and <2.8) of magnetically non-susceptible sand and quartz showed that the enzyme could be fixed on the supports. Malonic acid (16.3 nmol and 16.7nmol per g) appeared to be bound to alkylamine purified fraction of magnetically non-susceptible sand and alkylamine quartz, respectively. Studies on the effect of 6 M guanidine.HCl on immobilized trypsin demonstrated that immobilized trypsin had considerable stability against denaturation. The results obtained indicated that magnetically non-susceptible sand was found to be nearly as good as quartz for trypsin immobilization and that trypsin was covalently coupled to sand via 3-aminopropyltriethoxysilane and glutaraldehyde.

    • Microtubule associated proteins of goat brain

      A Majumder S K Banerjee P K Sarkar

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      The microtubule associated proteins of goat brain were separated from tubulin on the basis of their thermostability and then fractionated by chromatography on Sepharose 4B column. Analysis of the fractions by SDS-Polyacrylamide gel electrophoresis and assay of their tubulin-assembly-promoting activity indicate that this activity resides primarily in the tauproteins (mol. wt. 55,000–70,000) and a class of even lower molecular weight (25,000–35,000) proteins. Electrophoresis of the microtubule associated protein fractions separated from tubulin by phosphocellulose chromatography are in agreement with the results obtained from fractionation on Sepharose 4B columns.

    • Purification and properties of polyphenoloxidase of mango peel (Mangifera indica)

      T N Prabha M V Patwardhan

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      Polyphenoloxidase from mango(Mangifera indica) peel was purified to homogeneity by ammonium sulphate fractionation, chromatography on DEAE-Sephadex and gel filtration of Sephadex G-200. The enzyme had an apparent molecular weight of 136,000. Its pH and temperature optimum were 5.4 and 50‡C, respectively. The enzyme possessed catecholase activity and was specific too-dihydroxy phenols. The enzyme also exhibited peroxidase activity. Some non-oxidizable phenolic compounds inhibited the enzyme competitively. High inhibitory effects were also shown by some metal chelators and reducing agents, Mango peel polyphenol oxidase when immobilized onto DEAE Sephadex showed slightly higher Km for catechol and lower pH and temperature optima.

    • Isolation and characterization; of α-santonin assimilating pseudomonad

      U M X Sangodkar S Mavinkurve

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      Pseudomonas cichorii strainS, isolated by soil enrichment technique, utilized santonin as the sole source of carbon, forming chromatographically destinguishable transformation products. One of the intermediary transformation products was identified as 1,2-dihydro α-santonin.

    • Control of sporulation in the filamentous cyanobacteriumAnabaena torulosa

      Tonina A Fernandes Joseph Thomas

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      In the cyanobacteriumAnabaena torulosa, sporulation occurred even during the logarithmic growth phase. Sporulation was initiated by differentiation of the vegetative cell on one side, adjoining the heterocyst followed by differentiation of the vegetative cell on the other side. Subsequently, spores were differentiated alternately on either side to form spore strings. The sequence of sporulation supports the previous notion that a gradient of spore maturation exists in cyanobacteria and also indicates that the gradient is manifested unequally on either side of heterocysts. Sporulation was absent or negligible in a minerally enriched medium but ocurred readily in a minimal medium. The extent of sporulation was inversely related to phosphate concentration. Sporulation was enhanced at higher temperature. Incandescent light, but not fluorescent light, greatly stimulated sporulation suggesting possible involvement of red light in spore differentiation. Addition of filtrate, from 5 to 8 day old cultures, to freshly inoculatedA. torulosa greatly enhanced sporulation indicating the influence of extracellular products in spore formation.

    • Metabolism of glycosaminoglycans in rats during methionine deficiency and administration of excess methionine

      G Muraleedhara Kurup P A Kurup

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      Methionine deficiency in rats caused significant decrease in the concentration of many sulphated glycosaminoglycans in the aorta and other tissues, while administration of excess methionine caused an increase in these constituents. The activity of some important biosynthetic enzymes decreased in methionine deficiency and increased on administration of excess methionine. No uniform pattern was observed in the changes in the activity of enzymes concerned with degradation of glycosaminoglycans. The concentration of 3′-phosphoade-nosine 5′-phosphosulphate and the activities of the sulphate activating system and sulpho-transferase were decreased in methionine deficiency, while feeding excess methionine did not affect these parameters as compared to controls.

    • Properties and characteristics of an anti-human chorionic gonadotropin monoclonal antibody

      S K Gupta S Ramakrishnan G P Talwar

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      The product of a hybrid cell clone, P3W80, obtained as ascites fluid from mouse peritoneal cavity had high titres of anti-human chorionic gonadotropin antibodies e.g. 30 to 40% binding of125I-human chorionic gonadotropin at 107 dilution in a radioimmunoassay. The antiserum SB6 (raised against Β-human chorionic gonadotropin distributed by National Institutes of Health, USA gave similar binding at 5000 dilution in parallel runs. The monoclonal antibody recognized best human chorionic gonadotropin (0.3 mlU of hormone/tube withB/B0 75%), but also bound Β and a subunits of human chorionic gonadotropin, 12 and 800 folds lower than human chorionic gonadotropin respectively No binding was observed with carboxy terminal peptides of Β-human chorionic gonadotropin ranging from 93 to 145 amino acid residues, indicating the lack of recognition of the C-terminal region. No cross-reaction with human leutinizing hormone was obtained at the physiological surge levels, a significant competition (B/B075 %. obtainable only at 60 mlU of LER 960 human leutinizing hormone/ tube. The antibody had heavy chain of IgG1 and light chain of kappa type. It neutralized the bio-activity of human chorionic gonadotropin bothin vitro and invivo.

    • Analysis of bovidae genomes: Arrangement of repeated and single copy DNA sequences in bovine, goat and sheep

      Uma Mehra P K Ranjekar

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      Approximately 43–60% of the total genome in bovine, goat and sheep consisted of interspersed repeated and single copy DNA sequences. Most of the interspersed repeated DNA sequences were 1500–2400 nucleotide pair long while a minor portion was more than 4000 nucleotide pair long in goat and sheep and 3200 nucleotide pair long in bovine. About 1/3rd of single copy sequence were interspersed and their length was in the range of 1000–1500 nucleotide pairs.

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