Strain Improvement by Mutagenesis and Optimum Conditions for Culture Parameter by Response Surface Methodology for Lactose Tolerance in a Novel Native Culture Isolate Leuconostoc Mesenteroides Subsp Dextranicum

P. Shobha Rani, Renu Agrawal* and B. S. Ramesh

Large numbers of people all over the world are suffering from lactose intolerance. The present work was carried out to identify and enhance the enzyme activity in Leuconostoc mesenteroides subsp dextranicum, a native laboratory isolate deposited at MTCC, Chandigarh with accession No. 5209. The wild type strain had an activity of 7.5 mmoles/ mg protein. In the improved mutant strain the yield increased to 13.2 mmoles/ mg protein with ethyl methyl suphonate (EMS- 46.8 mg/l) treated and to 15.2 mmoles/ mg protein in UV (1 min) treated mutant. The UV strain was further studied for optimizing physical and chemical parameters by Response Surface Methodology (RSM) and the b-galactosidase activity was found to enhance 500 times (3920 mmoles/ mg protein) more when grown at pH 7.0 and supplemented with 1 % lactose at 600C. This high activity for b-galactosidase activity in Leuconostoc mesenteroides subsp dextranicum is being reported for the first time.

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Effect of Solvents and Stabilizers on the Preparation and Morphology of Insulin-loaded Poly-e-caprolactone Microspheres

Anindita Mukerjee and Vikas Pruthi*

Over the past few decades, there has been considerable interest in developing biodegradable microspheres as effective drug delivery systems. The characteristic function of these carrier particles is their ability to deliver drugs, especially proteins, across several biological barriers to the target site without denaturation and degradation. Our study focuses on the effect of various organic solvents and stabilizers and the optimum concentration of the stabilizers used for the preparation of smooth, spherical insulin-loaded poly-e-caprolactone microparticles. Results showed that dichloromethane was the solvent of choice and stabilizer (Hydroxypropyl methylcellulose) at a concentration of 1% yielded the best microsphere formation.

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Isolation, Properties and Growth Kinetics of A Bacterium Isolated From Bath Soap

K.L. Tiwari*, S.K. Jadhav and P. Shukla

A mesophilic, facultative anaerobic, spore-forming bacterium is isolated from a bath soap that was in regular use in bathroom. The isolated organism is rod shaped present in chains, gram positive, catalase and oxidase positive and actively motile in nature. Growth was observed at different pH ranging from 2.0 to 10.0 and the optimum growth was observed at pH 7.0. The optimum temperature for growth was around 45°C. Growth was observed with different ions out of which growth was present in Al, Mn, Zn, Ag, ferric and ferrous ions while absent in Cu and Co ions. The bacterium also grew well in detergents. Alcohol was produced by the bacterium.

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Partial Purification and Optimization of a Molecular Shark against Free Radicals – An Effective Approach

R. Kalpana, G. Vijayalakshmi and S. Adline Princy*

Catalase is an antioxidant enzyme produced in the body to scavenge hydrogen peroxide to water and oxygen, preventing the cell from oxidative damage. Rich plant sources of this enzyme include vegetables like tomatoes and potatoes. In our study, Kashmir apple was chosen for its nutritive value due to the presence of vitamins and apple polyphenols. As an initial part of our study, we tested our sample for a viable catalase source. After purification of catalase by ion exchange chromatography, its optimum properties were characterised. Maximum enzyme activity was found at 50°C and at a pH of 6. The purified enzyme had an activity of 9.7% and a specific activity of 92.18 U/mg of protein. Of the various inhibitors tested, copper sulfate and manganese chloride were found effective. This preliminary investigation provides an insight into its application part against the molecular sharks (free radicals).

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High Performance Liquid Chromatographic Quantification of Organic Acids and Glycerol in Rice Wine

M. Sundar*, S. Gunasekaran, M. Sukumar, M. Sivarajan and R. Rajesh Kumar

Rice wine is a drink made in Japan from fermented beverage that contains alcohol up to 16% and hundreds of rice wines are available across the world. Rice wines contain rich nutrients; they are good for bringing up a good appetite and good health upon regular drinking in suitable amounts. The organic acids present in wines were determined earlier by photometric, chromatographic and enzymatic methods but detection in HPLC is quite sensitive and selective method. The organic acids such as succinic, acetic and lactic acids and glycerol production of the different yeast isolates were analysed and quantified. The separation of succinic, acetic and lactic acids exhibited the retention times of 10 – 11, 8 – 9 and 6 – 7 min. respectively. Glycerol production was found in larger quantities compared to other organic acids. Among the various organic acids, succinic acid was produced in larger quantities by the yeast isolate YMs (2.68 mM) followed by the isolate YI (2.66 mM). In the case of acetic acid production, the yeast isolate YJ produced the maximum of 2.00 mM followed by YMs (1.42 mM). With regard to glycerol production, the highest production of 89.45 mM was obtained by the isolate YMs followed by YG. The values for succinic, acetic and lactic acids were all with low confidence limits in contrast to glycerol, which exhibited the highest values.

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A Comparison of ERA Rabies Strain Nucleotide and Deduced Amino Acid Sequences with the different Strains of Rabies Virus for Sequence Homology

N. Nagesha and P.H. Ramanjini Gowda*

Rabies is one of the most fearsome infectious diseases known to the animals and mankind. In our laboratory we have developed the transgenic crops with rabies glycoprotein gene using ERA (Evelyn Rokitnicki Abelseth) rabies virus strain. The molecular techniques confirmed the integration and expression of the rabies glycoprotein in the transgenic crops. In order to test whether the transgenic crops expressing glycoprotein of ERA strain can prevent the rabies disease caused by the other strains, we have carried out comparison of ERA nucleotide and amino acid glycoprotein sequences with different strains of rabies virus. The different strains used for the comparison are CVS, Fleury, PM 1503, PG, Chinese etc, using the CLUSTAL W multiple sequence alignment. More than 90 per cent of the homology was found across the different strains, which indicate that the glycoprotein coding sequences of different strains are highly conserved across the rabies strains.

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Antifungal Effect of L. plantarum on Spoilage Moulds in ‘Ogi’

Obadina A.O* and Oyewole O.B

Sixty-four strains of Lactobacillus plantarum were isolated during the four days of maize fermentation. Moulds involved in the spoilage of cooked fermented maize gruel (ogi) were also isolated. These moulds include Aspergillus niger, Aspergillus flavus and Neurospora sitophila. The effects of the Lactobacillus plantarum strain on the moulds were investigated. Lactobacillus plantarum strain was effective against the moulds with highest effect on Neurospora sitophilla followed by Aspergillus niger and Aspergillus flavus. L. plantarum strains OA9605 exhibited the highest antifungal activity. The effect of crude L. plantarum extracts on the cooked liquid gruel (ogi) was studied. The ‘ogi’ inoculated with L. plantarum extracts had a shelf life of 6 days. The antifungal substance was heat table.

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Potential Application of Fungal Biotechnology on the Nutritional Evaluation of Saw Dust-Gliricida Mixture by Rat

Belewu M. A.*, Aderolu Z. A., Banjo M. O., Musa A.K., Oyerinde A. A. and Andsalami O.S.

The biological and chemical processing methods on Sawdust - Gliricidia mixtures were examined in a completely randomized design model. The Saw dust - Gliricidia mixture was treated chemically with 50g of 25% NaOH on 1kg of Sawdust - Gliricidia mixture while the biological method was by incubation of Sawdust - Gliricidia mixture with mushroom (Pleurotus sajor caju). Both methods are used with the aim of achieving degradation and solubilization of fibre fractions (cellulose, lignin and ADF) while its nutritional importance was evaluated using thirty (30) Albino rats. The results revealed significance differences in the ADF, lignin, cellulose, CF, CP, EE and ash of the control (A^ untreated Saw dust - Gliricid-mixture), biological (B) and chemically treated sample (C). The degradation and solubilization of the fibre fraction were more pronounced in the biological (mushroom) treated sample compared to the control and chemically treated samples. Dry matter intake of diet B was higher than A and C by 20.99 and 35.76% respectively while the crude protein intake was 46.30 and 51.92% respectively. The crude fibre intake was lowest for C, B and A in that order. Apparent digestibilities of CP, CF, ADF and lignin were increased (p<0.05) for the biological treated (diet B) sample. The rate of gain was significantly (P< 0.05) higher in the biological treated sample compared to the other two samples. It is concluded that the biological treatment of Sawdust-Gliricidia mixture mimics the effect of degradation of solubulization of fibre fraction and it will be a useful method for pre-digesting lignocellulose materials for possible rumen microbial attack

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Influence of Growth Conditions on the Production of Bacteriocin by Lactic Acid Bacteria Isolated from Camel’s Milk

Salha H. Al-Zahrani* and Fozyah S. Al-Zahrani

Fifty six isolates of wild lactic acid bacteria (LAB) were isolated from eleven samples of camels΄ milk. All isolates were belonging to coccal shaped, CJ1 and CS2 belonged to Pediococcus sp. while CS3 and CF4 belonged to Lactococcus lactis ssp lactis. They showed antagonistic activities against spoilage and food-borne pathogenic bacteria, Salmonella typhi, Staphylococcus aureus and Lesteria monocytogenes as indicators. Four isolates with the highest antagonistic activities, CJ1, CS2, CS3 and CF4 were chosen for more studies. Inhibition due to hydrogen peroxide, organic acids were ruled out, so the antagonistic activities against Staphylococcus aureus can be described to the production of bacteriocin. The effect of incubation period, temperature, pH value and different media on the production of bacteriocin were studied. Production of bacteriocin in MRS broth was found to be higher than in M17 and M17L broth. It was found that bacteriocins were not destroyed at temperature 4ºC for 72h and at 30ºC, 80º , 100ºC for 30min and even at 121°C for 15 min. The activity was maintained at pH of 2.5-4.5 or 2.5-5, depending on the LAB strain. The bacteriocins were inactivated by the addition of proteinase K, but that produced by isolates CJ1and CS3 was sensitive to tyrosine. However, bacteriocins produced by isolate CS2 and CS3 were sensitive to pepsin. The activities of bacteriocins were not destroyed but it became lower by adding 40-65% of NaCl.

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Isolation, Properties and Growth Kinetics of A Bacterium Isolated From Bath Soap

K.L. Tiwari*, S.K. Jadhav and P. Shukla

A mesophilic, facultative anaerobic, spore-forming bacterium is isolated from a bath soap that was in regular use in bathroom. The isolated organism is rod shaped present in chains, gram positive, catalase and oxidase positive and actively motile in nature. Growth was observed at different pH ranging from 2.0 to 10.0 and the optimum growth was observed at pH 7.0. The optimum temperature for growth was around 45°C. Growth was observed with different ions out of which growth was present in Al, Mn, Zn, Ag, ferric and ferrous ions while absent in Cu and Co ions. The bacterium also grew well in detergents. Alcohol was produced by the bacterium. In Focus

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Application of Microwave Irradiation in Biocatalysis

Sanjib Kumar Karmee

This feature focuses on effect of microwave irradiation on enzyme catalyzed reactions.

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Edible Vaccines from Transgenic Plants

Virupakshagouda U. Patil* and Dharmendra P. Patil

Vaccines have accomplished miracles in the fight against infectious diseases. They have consigned smallpox to history and should soon do the same for polio. Currently two forms of vaccines are available-one purified from serum of infected individuals and other a recombinant antigen expressed and purified from yeast. Almost all current vaccines are injected. As a result, several problems arise, such as viability and cost of skilled labour to perform the injection, the cost of producing injected vaccines and the need for cold storage. However, there are alternatives to injection as the means for vaccination, mucosal immunization is one such option. Dr. Charles Arntzen of Arizona State University has put the idea of stable, plant-based vaccines as an alternative. Plant based vaccines can be grown locally, reducing the cost and complications of transportation, while the stability of proteins in intact plants removes the need for refrigeration furthermore, the edible nature of vaccines eliminates the need for syringe-based delivery, saving money and reducing the risk of infection. Till date edible vaccines are produced against many diseases of both human and animal, some of these important are discussed with reference to the present contest.

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The Rudiments of Enzyme Immobilization

Sanjib Kumar Karmee

Immobilized enzymes are economical than its native counterpart as they are reusable and possess the advantage to be used in continuous bioprocess. There are numerous method reported in literature for immobilizing a biocatalyst. However, it is important to choose an immobilization method, which avoids reaction of the matrix with amino acid residues present in the active site of an enzyme. Immobilization often stabilizes the enzyme-conformation. Hence, increases their stability and application in non-aqueous media. In this article the fundamentals of immobilization are discussed which includes: definition of immobilization and immobilized-biocatalyst, advantages of immobilization and different methods of immobilization. This article also gives an insight look in to the sub-classes of each immobilization method.

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