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Rishīkesh, India

Jain R.,Banasthali University | Garg V.,Banasthali University | Dangwal K.,Modern Institute of Technology | Lily M.K.,Modern Institute of Technology
Bioscience, Biotechnology and Biochemistry | Year: 2013

The present study aimed at a comparative characterization of two distinct extracellular monocrotophos hydrolases, from Penicillium aculeatum ITCC 7980.10 (M3) and Fusarium pallidoroseum ITCC 7785.10 (M4), isolated from agricultural fields. The MCP hydrolases were purified by Sephadex G-100 column and DEAESepharose CL-6B ion-exchange column followed by SDS-PAGE analysis, which showed the presence of two hydrolases, of 33 and 67 kDa respectively. Both enzymes were most active at alkaline pH and were stable over a wide range of temperatures (60-70 C). Between the strains, the MCP hydrolases from M3 were 2-fold more active than that from M4. Enzyme kinetic studies showed lowest Km (33.52mM) and highest Vmax (5.18 U/mg protein) for OPH67 of M3 in comparison to the Km and Vmax of the other hydrolases purified from M3 and M4, suggesting that M3 OPH67 was the most efficient MCP hydrolase. To the best of our knowledge, this is the first report of the purification of two distinct extracellular thermostable MCP hydrolases from fungal strains Penicillium aculeatum ITCC 7980.10 and Fusarium pallidoroseum ITCC 7785.10. Owing to its potential MCP hydrolyzing activity, M3 OPH67 can perhaps used directly or in the encapsulated form for remediation of MCP contaminated sites. Source


Objective: The present study aimed the characterization and purification of acid phosphatases from monocrotophos (MCP) hydrolyzing Aspergillus niger ITCC 7782.10 isolated from local agricultural field. Material and Methods: Intracellular phosphatases from MCP hydrolyzing Aspergillus niger ITCC 7782.10 were purified by four step purification procedure which were further characterized. Results: The study demonstrated the presence of two intracellular phosphatases of varying molecular weights of 33 and 67 kDa which were purified to 126 and 76 fold respectively by four step purification strategy. Both the phosphatases showed similar pattern for pH, temperature and metal ion however, the activity of P33 was found to be higher than that of P67. The phosphatases were found to be most active at acidic pH and were found to be stable till 80°C demonstrating them to be acid phosphatases and thermos table. Km and Vmax values of P33 was found to be 0.28 mM and 1.21 Umg-1protein respectively whereas Km of P67 was found to be 0.72 mM and Vmax value was 1.85 Umg-1protein. This indicated that P33 had more affinity towards para nitro phenol phosphate (pNPP) as a substrate than P67. Conclusion: This is the first report indicating the presence of two different intracellularly thermostable acid phosphatases within the same MCP hydrolyzing fungal strain Aspergillus niger ITCC 7782.10. Therefore, these acid phosphatases could be effectively used for the bioremediation of MCP contaminated sites. Source


Singh H.,Modern Institute of Technology | Lily M.K.,Modern Institute of Technology | Dangwal K.,Modern Institute of Technology
Asian Pacific Journal of Tropical Disease | Year: 2015

Objective: To evaluate and compare the polyphenol contents, antioxidant, anti-elastase, anti-collagenase, anti-tyrosinase and anti-inflammatory activities of 13 wild edible fruits [Pyracantha crenulata, Berberis asiatica (B. asiatica), Ficus subincisa (F. subincisa), Morus serrata, Ziziphus nummularia, Leea asiatica (L. asiatica), Dendrobenthamia capitata, Ziziphus mauritiana, Prunus cerasoides, Ampelocissus latifolia (A. latifolia), Vitis jacquemontii, Morus alba and Grewia optiva] of North-West Himalayan Region of India. Methods: Fruits extracts were prepared with 80% aqueous acetone and evaluated for total phenolic contents (TPC) and total flavonoid contents (TFC). Free radical scavenging activities [against 1,1-diphenyl-2-picryl-hydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), linoleate hydroperoxyl and superoxide radicals], ferric reducing ability, ferrous metal chelating capacity, anti-elastase, anti-collagenase, anti-tyrosinase and anti-inflammatory activities were determined by using various in vitro assays. Results: TPC varied from 58.83 to 4 496.39 mg gallic acid equivalents/100 g fruit weight (FW), being highest in A. latifolia and lowest in F. subincisa. TFC ranged from 108.00 to 1 963.75 mg catechin equivalents/100 g FW, standing highest in L. asiatica and lowest in Prunus cerasoides. A. latifolia and L. asiatica possessed the highest antioxidant activities while B. asiatica and L. asiatica owned uppermost anti-elastase and anti-collagenase activities, respectively. B. asiatica revealed the highest anti-tyrosinase activity and F. subincisa demonstrated the highest anti-inflammatory activity. The present study revealed differential contribution of TPC and TFC in various antioxidant activities. However, no obvious relationship was visible between anti-elastase/anti-collagenase/anti-tyrosinase/anti-inflammatory activities and TPC/TFC, suggesting the role of individual or combination of specific phenolics and flavonoids. Conclusions: The abilities of Himalayan wild edible fruits to scavenge a variety of free radicals, inhibit enzymes causing skin-aging and skin-darkening are highly appreciable, suggesting their possible utilization for the development of effective formulations for general health maintenance and anti-aging, skin-whitening cosmetics. © 2015 Asian Pacific Tropical Medicine Press. Source


Objective: The present study reports for the first time the production, purification and characterization of α-amylase from a known HMW-PAHs degrader Bacillus subtilis BMT4i. Methods: Culture conditions for the production of α-amylase were optimized. The α-amylase was further purified partially by ammonium sulphate precipitation and kinetic characterization of the α-amylase was done. Results: The observations demonstrated BMT4i as an efficient producer of α-amylase. that revealed maximum production at 72 h, pH 8.0, starch (20 g/l), peptone (10g/l) and CaCl2(0.2g/l). The α-amylase exhibited a specific activity of 1001.08 U/mg corresponding to 3.86 fold purification and 76.7% yield. The enzyme exhibited optimal activity at 40°C and pH 8.0. The enzyme was stable in the pH range of 4.0-8.0 and retained stability at 50°C for 2 h. The Vmaxand Kmof α-amylase was found to be 5000 U and 4.0 mg ml-1 respectively. The enzyme activity was strongly activated by Ca2+and Fe3+. Conclusion: Our findings emphasize upon the prospect of the commercial production of α-amylase from Bacillus subtilis BMT4i that can be employed in various sectors such as food, pharmaceuticals, textiles, detergents, etc. © TurkJBiochem.com. Source

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