Abstracts
BGS-Haridrin : A glycoprotein from boiling water extract of Turmeric (Curcuma longa L)
Dinesha R. and Leela Srinivas
Adichunchanagiri Biotechnology and Cancer Research Institute,
Balagangadharanatha Nagara-571448, Mandya District,
Karnataka, INDIA
Corresponding Author
Dr. Leela Srinivas, Ph.D.
Adichunchanagiri Biotechnology and Cancer Research Institute,
Balagangadharanatha Nagara-571448, Mandya District,
Karnataka, INDIA
Email: directorabcri@yahoo.com
BGS-Haridrin - a glycoprotein isolated from boiling water extract of Turmeric (Curcuma longa L). The extract of turmeric gave three peaks (P-I, P-II and P-III) in Sephadex G-25 chromatography. P-I showed antioxidant activity and hence subjected to DEAE A-25 chromatography showed five peaks. Peak IV showed antioxidant activity, this was named as BGS-Haridrin. SDS-PAGE and MALDI/MS analysis showed its molecular weight as ~28 kDa. BGS-Haridrin at 0.9 nM concentration shows 82%, 80%, inhibition in hydroxyl radical scavenging and lipid peroxidation inhibition assay respectively. BGS-Haridrin provides protection against H2O2, t-BOOH induced DNA damage and also against Beedi, Cigarette and Cow dung smoke condensate carcinogens and reactive oxygen species induced DNA damage. Our protein showed antibacterial activity against strains like Staphylococcus aureus, Proteus vulgaris, Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium, Vibrio cholerae, Streptococcus sps and Pseudomonas sps. MIC values of BGS-Haridrin (3.25µM) was eight time lesser than Streptomycin (25.8 µM). BGS-Haridrin was also found to be effective against the fungi Candida albicans. MIC of BGS-Haridrin (1.5µM) was 16 times lesser than Amphotericine (23.8µM).
Thus, the present study demonstrates the in vitro antioxidant, DNA damage protectant activity and antibacterial of BGS-Haridrin.
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ANTIMICROBIAL AND ANTIPLASMODIAL ACTIVITY OF ARISHININ: AN ANTIOXIDANT PEPTIDE ISOLATED FROM CURCUMA LONGA LINN.
Harsha Ramakrishna and Leela Srinivas
Adichunchanagiri Biotechnology and Cancer Research Institute (ABCRI),
Balagangadharanatha Nagara-571448, Mandya District,Karnataka, INDIA.
Corresponding Author
Dr. Leela Srinivas. Ph.D.
Director, Adichunchangiri Biotechnology and Cancer Research Institute
Balagangadharanatha Nagara - 571448
Nagamangala Taluk, Mandya, Karnataka, INDIA.
Tel: +918234-287850
Fax: +918234-287984
Email: directorabcri@gmail.com
Arishinin, an antioxidant peptide (Mol wt:~8.2kDa) isolated from boiling water extract of turmeric rhizome (Curcuma longa linn). It is an effective ROS quencher and at 0.73µM it displays 87 and 85% inhibition in TBARS and hydroxyl radical induced oxidative stress. Against DPPH and Nitric oxide free radicals, at 1.87µM, it showed 82 and 89% inhibition respectively and on a comparative scale to the standard antioxidants like Curcumin, a-Tocopherol and BHA, arishinin was required only at 10-20 times lesser dose. Antimicrobial activity by Disc diffusion and MIC value by Serial dilution was done against microbes viz E. coli, V. cholera, K. pneumonia, S. aureus, S. typhimurium, Pseudomonas, P .vulgaris, C. albicans, A. flavus, T .mentagranophytes and Rhizobium. Streptomycin and Amphotericin-B was used as standard antibacterial and antifungal agents. Average MIC values of Arishinin against bacterial and fungal strains were 2.5 and 6µM and it was 10 and 5 times lesser than the standard antibiotics like Streptomycin and Amphotericin-B. Against the malarial vector, Plasmodium falciparum, Arishinin was effective on both Chloroquine-resistant RKL-9 and Chloroquine-sensitive MRC-2 strains with an EC50 of 24 and 27nM where as Curcumin showed 50% inhibition at 5µM concentration. Arishinin was 208 and 185 times better than Curcumin against CQ-res and CQ-sen strains of P.falciparum respectively.
Arishinin, a peptide isolated from Curcuma longa is a novel Antioxidant, Antimicrobial and Antiplasmodial non toxic peptide with a possible preventive/curative health benefit.
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Isolation and characterization of hemostatically active components from cold water extract of Curcuma longa Linn. rhizome
Corresponding Author
Dr. Leela Srinivas. Ph.D.
Director, Adichunchangiri Biotechnology and Cancer Research Institute
Balagangadharanatha Nagara - 571448
Nagamangala Taluk, Mandya, Karnataka, INDIA.
Tel: +918234-287850
Fax: +918234-287984
Email: directorabcri@gmail.com
Traditionally in India turmeric is applied on cuts to stop the bleeding. The crude cold water extract of Curcuma longa Linn. exhibits proteolytic activity, as estimated by the caseinolytic assay. Casein hydrolyzing activity of a protease was inhibited by PMSF indicating that the protease possibly belongs to serine protease family. The crude extract also exhibits pro-coagulant activity by reducing plasma clotting time from 235 to 66s and promotes blood coagulation. It also hydrolyses human fibrinogen Aa and then the Bß subunits, whereas ? subunit is resistant. The extract also shows plasmin like activity as it hydrolyses fibrin clot subunit which is cross linked a-polymer and the partial hydrolysis of ? - ? dimer. The a and ß chain of fibrin appears to be resistant. The hydrolysis of human fibrinogen and fibrin clot by crude rhizome extract implies significant hemostatic property in relevance to wound healing. This study gives the scientific credence to C. longa with its propensity to stop bleeding and wound healing. However it is noteworthy that the boiling water extract of C. longa containing the antioxidant protein-Turmerin of 14 kDa (UNIPROT P-85278) does not show any protease activity and on the other hand, it is resistant to pepsin and trypsin.
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ANTIPROTEASE ACTIVITY OF TURMERIN ISOLATED FROM AQUEOUS EXTRACT OF TURMERIC
Prabhu MSL, Chethan Kumar M, Sampath Kumar S, Leela Srinivas
Adichunchanagiri Biotechnology and Cancer Research Institute
BG-Nagara, Nagamangala Taluk, Mandya District-571448, India.
Corresponding Author
Dr. Leela Srinivas. Ph.D.
Director, Adichunchangiri Biotechnology and Cancer Research Institute
Balagangadharanatha Nagara - 571448
Nagamangala Taluk, Mandya, Karnataka, INDIA.
Tel: +918234-287850
Fax: +918234-287984
Email: directorabcri@gmail.com
BGS-Turmerin(14kDa) is a water soluble colourless protein isolated from the boiling water extract of turmeric powder (Curcuma longa Linn). The purification of BGS-Turmerin was done by Sephadex G-50 chromatography which yields the antioxidant protein-Turmerin which is partially sequenced and the sequence is LCPLDVLQLS-SELLDIDGNE-VEASRILSDI-TAFGGIRCPL-TVVQSRGIGT-IISSPYRFIA, and is registered with UNIPROT registration number P-85278. This is a protease inhibitor like Kunitz of Soyabean and Miraculin of Miracle plant (Synsepalum dulcificum). It inhibits the protease activities up to 77% at 180nM dose of Russell's viper venom (Daboia russellii russellii) which has both serine and metallo proteases and also of Naja naja venom containing serine proteases up to 80% at 180 nM concentration. The above proteases are major hydrolytic enzyme responsible for the early effects of envenomation such as hemorrhage, pro-coagulation of blood and tissue damage. BGS-Turmerin prolongs the venom induced plasma clot formation by protecting the degradation of Aa and Bß chains of human fibrinogen thus providing precious minutes of life of the snake bite victim. Thus, BGS-Turmerin is a unique novel antioxidant protein, resistant to trypsin and pepsin and can withstand heat treatment up to 1000 C for 180 minutes and the biological availability is approximately 70%.
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Little is beneficial: Curcumin with potential for implantation
Sumit Paul, Snehasikta Swarnakar
Department of Physiology
Indian Institute of Chemical Biology
Kolkata-700 032, India.
Corresponding Author
Dr. Snehasikta Swarnakar, Ph. D.
Head, Department of Physiology, Indian Institute of Chemical Biology
4, Raja S.C. Mullick Road, Jadavpur, Kolkata-700032, India.
Tel: 91-33-2473-0492 Ext 224; Fax: 91-33-2473-5197.
Email: snehasiktas@hotmail.com
Although, curcumin, derived from turmeric, notably heals most pathologic conditions but we understand very little about it. It is high time to unravel the specificity of this important molecule to minimize the mistake. Implantation in general and embryo implantation in specific is one of the important gateway to study embryogenesis as well as tumorogenesis. We investigated whether curcumin, the old common spice of Indians, has any effect on implantation of blastocyst on to uterus in mice that are exposed to arsenic. Arsenic toxicity from consumption of ground water is a menace in the eastern parts of India with severe consequences in West Bengal.
Arsenic prone areas also show high child mortality, defected and aborted foetuses. Under acute arsenic toxicity, mice showed implantation failure. Curcumin was found beneficial in embryo implantation at very low dose ( <25 mg/kg b w for 3 consecutive days during pre implantation period) in mice exposed to arsenic toxicity, while excess doses caused detrimental effect to mice under similar situation. The study has the potential to open up a critical issue for judicial use of curcumin. Thus, turmeric exerts specific role on different biochemical pathways and advises against its random consumption especially for would be mothers.
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Curcumin arrests gastric ulcer under diabetic milieu
Laishram Pradeepkumar Singh, MSc.
Anamika Vivek Sharma, PhD and Snehasikta Swarnakar, PhD
Indian Institute of Chemical Biology
4 Raja S C Mullick Road, Jadavpur, Kolkata-700032.
Email: snehasiktas@hotmail.com
Fax: 033-2473-5197.
Gastric mucosa in diabetic mellitus becomes vulnerable to various ulcerogens leading to severe ulceration. Diabetes mellitus, caused by insulin deficiency, is a metabolic disorder characterized by hyperglycemia that creates huge economic and psychological trouble on a large number of people of all ages, races and socio-economic classes throughout the world. Several studies have found association of reactive oxygen species, proinflammatory cytokines, dysregulated endogenous antioxidants etc. with the pathogenesis of diabetes and gastric ulcer in diabetic state. Curcumin (diferuloylmethane), a bioactive constituent from Curcuma longa possesses remarkable anti-inflammatory, antioxidant, and anticarcinogenic property.
We hypothesized whether curcumin can exert antiulcer effect in diabetic patients. We developed gastric ulceration in streptozotocin-induced diabetic rat to examine curcumin's action on gastroprotection. Our preliminary investigations indicated antidiabetic property of curcumin through lowering the increased blood glucose level in streptozotocin-diabetic rats and maintaining the decreased weight loss almost to the control rats. Furthermore, our results highlighted that curcumin protected indomethacin-induced gastric ulceration in streptozotocin-diabetic rats by downregulating the increased activity and expression of MMP-13. We conclude that antiulcer efficacy of curcumin for gastroprotection to diabetic as well as non-diabetic milieu. It is hoped that curcumin would be a better agent to rescue injury for diabetic people.
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The disease endometriosis is rescued by turmeric
Sumit Paul, Snehasikta Swarnakar
Department of Physiology
Indian Institute of Chemical Biology
Kolkata-700 032, India.
Corresponding Author
Dr. Snehasikta Swarnakar, Ph. D.
Head, Department of Physiology, Indian Institute of Chemical Biology,
4, Raja S.C. Mullick Road, Jadavpur, Kolkata-700032, India.
Tel: 91-33-2473-0492 Ext 224;
Fax: 91-33-2473-5197.
Email: snehasiktas@hotmail.com
We have examined the effect of curcumin, a constituent of turmeric, on surgically developed endometriosis in mice. Endometriosis is a gyneocological disorder characterized by growth of endometrium at site other than the uterus and involves aberrant matrix remodeling. Endometriosis with varying severity was developed in mice by peritoneal implantation of uterine fragments. Curcumin administered either post-endometriosis or pre-endometriosis could arrest the disease in a dose dependent manner. We have reported earlier regarding the significant upregulation of matrix metalloproteinase (MMP)-9 and downregulation of tissue inhibitor of metalloprotease (TIMP)-1 in endometriotic tissues while curcumin pre- and post-treatment rescue the changes during regression of endometriosis. Currently, we evaluated the changes in the activity of MMP-3 in endometriotic tissues with or without curcumin treatment.
The elevated activity of MMP-3 in endometriotic tissues was suppressed to control value in curcumin treated samples. In addition, the attenuated activity of MMP-3 was associated with increased expression of TIMP-3 suggesting the anti-inflammatory property of curcumin. Moreover, expression of angiogenic factor e.g. vascular endothelial growth factor decreased upon curcumin pretreatment in endometriotic tissues. We report here for the first time the anti-endometriotic property of curcumin via MMP-3 that may lead to new therapeutic intervention.
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