An extra cellular lipase was isolated and purified from the culture broth of Pseudomonas aeruginosa SRT 9 to apparent homogeneity using ammonium sulfate precipitation followed by chromatographic techniques on phenyl Sepharose CL-4B and Mono Q HR 5/5 column, resulting in a purification factor of 98 fold with specific activity of 12307.8 U/mg. The molecular weight of the purified lipase was estimated by SDS-PAGE to be 29 kDa with isoelectric point of 4.5. Maximum lipase activity was observed in a wide range of temperature and pH values with optimum temperature of 55ºC and pH 6.9. The lipase preferably acted on triacylglycerols of long chain (C14-C16) fatty acids. The lipase was inhibited strongly by EDTA suggesting the enzyme might be metalloprotein. SDS and metal ions such as Hg , Ag 2+ and Fe 2+ decreased the lipase activity remarkedly. Its marked stability and activity in organic solvents suggest that this lipase is highly suitable as a biotechnological tool with a variety of applications including organo synthetic reactions and preparation of enantiomerically pure pharmaceuticals. The Km and Vmax value of the purified enzyme for triolein hydrolysis were calculated to be 1.11 mmol/L and 0.05 mmol/L/min respectively.
According to International Diabetes Federation (IDF), India has 62.4 million people with diabetes and by 2030 it is predicted that the number will rise to 100 million. Studies claim that there are around 410 experimentally proven Indian medicinal plants which have anti-diabetic activity, of which the mechanism of action of 109 plants has been elucidated or reported. So, the need of the hour is to explore the claims of Indian medicinal flora and open up the facets of many Indian plants which are being examined for their beneficial role in diabetes. So, we created a database (InDiaMed) of Indian medicinal plants that captures their role in anti-diabetic activity. InDiaMed's features include chemical, pharmacological, biochemical and geographical information of the medicinal plant, scientifically relevant information of the plant, and the coherent research done on it in the field of diabetes. The database also includes the list of poly-herbal formulations which are used for treatment of diabetes in India.Availabilityhttp://www.indiamed.info
Insecticidal crystal proteins produced by Bacillus thuringiensis (Bt) are toxic to specific insect pests. They are used as alternatives to chemicals for controlling insect pests using transgenic plants and biopesticides. Three strains of Bacillus thuringiensis expressing cry1 and cry2 genes were identified in soil samples collected from different locations in India. The full-length cry1, cry1I, and cry2 type genes (3.5, 2.1, and 1.9 kb, respectively) were cloned and sequenced. The full-length clones of the three genes were sub-cloned in the E. coli expression vector, and expressed proteins were analysed. The expression of cry1, cry1I, and cry2 resulted in 132, 81, and 70 kDa proteins, respectively. The expressed proteins were tested against secondinstar larvae of Helicoverpa armigera. The expressed proteins of cry1 and cry2- type from BRI-86 exhibited the highest toxicity to H. armigera larvae with an LC50 of 2.34 and 0.2 µg/ml diet, respectively. The LC50 exhibited by the test proteins was two and four-fold lower than the reference Btk HD-1 proteins. Thus, the spore preparation or the genes can be used individually or in combination to control insect pests.
A paper-based strip was developed by the authors for the detection and quantitative determination of paracetamol in bulk and pharmaceutical formulations for point-of-care testing prior to the assay prescribed in the respective pharmacopeia. In the present paper, lab-on-chip technology was used for the assay of a simple NSAID drug paracetamol. The qualitative analysis of the drug on the strip was performed by using three different reagents such as sodium Meta vanadate (1%), potassium dichromate (0.01M), N-bromosuccinamide (1%). This method employs dissolution of the powdered sample of drug in distilled water, followed by placing the sample solution of the drug on the strip of paper which was previously impregnated with the reagent. Drug concentrations were quantified from color intensity images using a scanner in conjunction with image processing software MATLAB. Linear calibration curves were generated for the drug with each of the reagent. The results obtained were compared with standard methods such HPLC and found to be in accordance with standards. This technology provides qualitative analysis and a prior easy step before assay of the drug at appreciably reduced costs.
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