Research Journal of Biotechnology

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Formulation of a Natural Medium for the Induction of Callus and their Bioactivity in Alstonia Venenata R. Br

Williams Scott J. and Thankamani V.

Preparation of an appropriate tissue culture medium was attempted to induct callus formation using explants from Alstonia venenata R.Br, Apocynaceae, an important medicinal plant used by folklore especially against snake venom. Phyto-hormones used conventionally in standard tissue culture media were substituted by natural growth supplements such as tender coconut water, coconut milk and tomato juice to determine their influence on growth and callus induction. Surface sterilization was optimized with 3 % (v/v) detergent ‘Triple 100’ (Rankem, India) for 20 min. followed by 0.2% HgCl2 (w/v) for 4 min.There was no contamination in any sample. Phenolic exudation was highly reduced through the use of natural additives. Node, internode, leaves and shoot tip were used as explants. The optimal concentration of both coconut water and coconut milk for callus induction was 30%. Tomato juice had no effect on callus induction at any concentration (10 to 30%) but phenolic exudation was reduced at 20%.Tender coconut water was found to be the best forming friable brownish white internodal callus with rhizogenesis in Murashige & Skoog medium after 1 month of incubation at 25° ± 2°C, 60 % humidity and in complete darkness. Coconut water was also favorable in forming brownish white hard embedded calli in contrast to brownish white calli formed on the surface of the media with coconut milk. It is the root bark in particular that is used against snake bites.Natural mode of propagation is rarely through seeds and hence the need for mass propagation by cell culture, induction of callus and rhizogenesis and extraction of bioactive compounds. Callus induction and rhizogenesis in Alstonia venenata using easily available cheaper natural additives is reported here for the first time to the best of our knowledge based on literature.The callus extract showed antimicrobial activity against bacterial pathogens. Production of secondary metabolites as an effective and potent antidote against snake venom and design and development of 100% effective drugs could be a near possibility.

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Rotation of F1-ATPase with a Truncated Rotor in the presence of Lauryldimethylamine Oxide

Hossain Mohammad Delawar1,2* and Ali M. Yusuf2

A continual supply of energy is required for living things to operate all biological processes which keep them alive. Some biological energy is stored in the adenosine tri-phosphate (ATP) molecules. ATP releases this energy in many useful ways in the cell and ATP is synthesized in the catalytic sites of F1-ATPase. F1-ATPase is an ATP-driven motor protein in which the central g subunit rotates with respect to the surrounding a3b3 cylinder. In our previous studies1, 2 we showed that mutant F1-ATPase made by deleting 14 to 21 amino acid residues from the carboxy terminus of the g subunit exhibited complicated hydrolysis kinetics and they could generate half or more torque compared to the wild type. Complicated hydrolysis kinetics of mutants became simple in the presence of lauryldimethylamine oxide (LDAO). Here we compare the effect of lauryldimethylamine oxide (LDAO) on the rotary behavior by estimating torque generated by the mutants and wild type in the presence of LDAO. Rotation of single molecules of F1 was observed under a microscope using 0.49 mm bead duplex as a marker at 2 mM ATP. The experiment was performed at room temperature (23 0C) and using an ATP regenerating system. The study shows that torque produced by the wild type or mutants remains unaffected upon addition of 0.15% LDAO.

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Physico-Chemical Analysis of River and Aquaculture Pond Water from Bhitarkanika Mangrove Ecosystem of Orissa

Mishra R. R., Thatoi H. N.*, Rath B.1, Mishra R. K. and Mohanty R. C.2

Physico-chemical parameters like temperature, pH, electrical conductivity, dissolved oxygen, dissolved carbon dioxide, chloride, magnesium hardness, calcium hardness, nitrate nitrogen, phosphorous and chloride contents of two water bodies i.e., the river water (lotic) and the aquaculture pond (lentic) in Bhitarkanika mangrove ecosystem of Orissa were analyzed during June-August 2005. The river water bodies are affected by industrial, agricultural and domestic pollutants which make their way into river. The most dominant among them are organo-pollutants such as fertilizers and pesticides used in agricultural fields, which are drained into the river during rainy season through surface runoff. The aquaculture ponds are also affected by human activities including application of medicine, feed and lime. Apart from this, excreta of fish and growth of microorganisms may also affect the pond water to a great extent. Both the quality of river and aquaculture pond water are also under influence of fresh water flow and the tidal inundation. The present paper reveals significant differences in the concentrations of various parameters between the two water bodies, which vary with monthly studies.

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Studies on Chemical Modification of Crotalaria Pallida Lectin

Hamid Rabia and Masood Akbar

Group specific chemical modification of lectins has been used as a tool to probe the nature of residues involved in lectin activity and to study the conformational changes induced by such modifications. A lectin isolated and purified from Crotalaria pallida seeds was chemically modified using acetic anhydride, succinic anhydride and iodoacetic acid. Fifty and ninety percent derivatives of each of the modified preparations were homogenous with respect to size as judged by size exclusion chromatography. Conformational changes were investigated by gel filtration, viscometery and ultraviolet absorption spectroscopy. Hydrodynamic properties were studied by gel filtration on a calibrated Sephadex G – 200 column and by viscosity measurements. Results from both these techniques indicated significant increases in hydrodynamic volumes in succinylated and acetylated lectins but no significant changes in carboxymethylated lectins. A similar behaviour was observed in the intrinsic viscosity values of the native and the modified lectins. Carboxymethylated lectin behaves like the native protein in most of its physicochemical properties.

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Variations in Serum Lipids and Thyroid Hormone Levels during Various Phases of Menstrual Cycle in Healthy Females

Pahwa Manju Bala* and Seth Shashi

The present study was carried out to evaluate comprehensively the inter relationship and impact of thyroid hormones on cyclical variation of serum lipid levels during menstrual cycle. Serum lipid fractions and thyroid hormones were measured in a total of thirty female healthy volunteers by commercially available kits in the Biochemical Laboratory, PGIMS, Rohtak. Total serum cholesterol, LDL-cholesterol and triglyceride levels significantly increased (p<0.05) in follicular phase compared to luteal phase. Serum T3, T4 levels decreased in follicular phase insignificantly (p>0.05). Serum HDL-cholesterol increased 3mg/dl or 5% in follicular phase of menstrual cycle. Insignificant decrease in thyroid hormone levels with significant increase in serum lipid fractions during follicular phase, warrant further investigation in this field. Moreover, phase of menstrual cycle should be taken into account whenever evaluating premenopausal women for plasma lipid abnormalities.

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Effect of Bt gene in intra hirsutum Bt hybrids of cotton

Ahuja S.L.

Estimates of heterosis over three commercial checks and correlation between the heterosis of seed cotton yield and lint yield with their component traits were compared in 8 Bt and corresponding non-Bt hybrids developed by private research and development sector of India. Eight Bt differed significantly from each other like non-Bt hybrids for all the characters. Bt hybrids could produce higher seed cotton yield, lint yield, heavier and higher number of bolls but could not excel their counter parts in lint percent, desirable plant height, earliness and fibre quality traits. None of the Bt and non-Bt hybrids could produce heavier bolls and seed and was earlier than the commercial hybrid check ‘LHH-144’. Heterosis of lint yield and seed cotton yield exhibited negative correlation with boll number /plant, number of sympodial branches and lint percent and days to flowering and plant height gave positive association in comparison to their counterparts. In Bt hybrids, micronare value and fibre strength heterosis in comparison to their counter parts depicted positive associations with lint yield and seed cotton yield indicating possibility of simultaneous improvement of the two traits.

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Biosorption of Chromium (Iii) Uptake using Immobilized Brevibacterium Species

Kishore Kumar K., Krishna Prasad M., Sarma G.V. S. and Murthy Ch. V. R.

Studies on removal of chromium using Biosorption technique have indicated that considerable amount of chromium can be removed using Brevibacterium species. Four different types of Brevibacterium species (Brevibacterium divaricatum NCIM 2634, Brevibacterium immariophilum NCIM 2237, Brevibacterium roseum NCIM 2270, Brevibacterium saccharolyticum NCIM 2238) were immobilized and used as adsorbents for the removal of chromium. The results show that the uptake of chromium was 62.4% in the absence of bacteria, 95.72% to 99% in the presence of Brevibacterium species. The adsorption capacities were found to be in the order Brevibacterium roseum < Brevibacterium saccharolyticum < Brevibacterium divaricatum < Brevibacterium immariophylum. The equilibrium data were well correlated with Freundlich type of adsorption isotherm. Finally empirical correlations were developed to estimate adsorption equilibrium.

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Fermentative Production of the Extracellular Exo-acting Inulinase from a Novel Strain of Erwinia sp.

Abraham Amith and Jayachandran K.

A novel strain of Erwinia sp. isolated from soil was capable of producing maximum amount of inulinase in a medium supplemented with 1% sucrose and 0.2% inulin at optimum conditions of incubation period 18 hours, pH 6 and temperature 300C. Evaluation of the influence of inulin on the growth of the organism indicated that inulin in the medium was used as an inducer and 0.4% was found to be the optimum inulin concentration for maximum enzyme production. The enzyme produced in an automated laboratory fermentor under the optimum conditions was subjected to purification by ammonium sulphate, dialysis and ion exchange chromatography. The exo-acting nature of the inulinase enzyme was proved by thin layer chromatography.

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Review Paper: Bacterial Lipases: Classification, Properties and Applications in Biotechnology

Joshi G.K. and Sharma Vinay

The study of lipases is a subject of special interest, because it lies on the borderline where the biological and physical sciences meet. In addition, lipases are the most used enzymes in synthetic organic chemistry, catalyzing hydrolysis of fats or oil or the reverse reaction in organic solvents. The contribution of bacteria and fungi to microbial biotechnology for production of lipases has far exceeded those of the other classes. Bacteria as a group are more versatile than fungi. In the range of organic compounds it can use as sole carbon and energy sources. The seemingly limitless diversity of bacteria provides an immense pool of raw material for novel lipase production. Keeping in view of the importance of bacterial lipases in this review paper, the classification, production and utility of these remarkably versatile biocatalysts have been discussed.

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Proteomics: Advanced Technology for Understanding Complex Traits of Molecular Biology

Mushtaq Roohi1 and Zargar Sajad M.2*

Proteomics is the study of the function of all proteins expressed by the genetic material of an organism at a particular moment during its life cycle. 2D electrophoresis and Mass Spectrometry are two important technological aspects for protein studies, which have been well exploited. Tremendous progress has been made in the past few years in generating large-scale sequencing data sets. The utilization of this sequence data is needed for exploiting the function of various genes of any organism under specific conditions. Proteomics is the recently emerged most powerful tool that can drastically help in understanding the biochemical and physiological mechanisms of complex traits at the molecular level. The discipline of proteomics has been initiated to complement physical genomic research. In this review, focus has been made on the rice proteomics studies as it has most extensive proteomic coverage to date.

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