Faculty Publications
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Item Coconut coir pith, available in abundance especially in tropical countries, was studied as a substrate for the production of cellulase[1,4(1,3;1,4)???D?glucan 4?glucanohydrolase, EC 3.2.1.4] and ??D?glucosidase(??D?glucoside glucohydrolase, EC 3.2.1.21) in solid state fermentation. The effects of fermentation time, nutrient level, substrate particle size and inoculum size have been examined for optimal production of these enzymes by the fungal strain Aspergillus niger NCIM 1005. The highest filter paper activity (FPA) of 4.11 IU g?1, carboxyl methyl cellulose (CMCase) activity of 15·55 IU g?1 and cellobiase activity of 9·31 IU g?1 were obtained after 7 to 8 days of fermentation. Reese and Mandel's mineral solution in the substrate to mineral solution ratio of 1:10 (w/v) supported high cellulase and cellobiase activities. An inoculum size of 20–50% (v/v) based on the volume of mineral medium and substrate average particle size of 375 ?m were optimum for enzyme production. Copyright © 1994 SCI(Production of cellulases from coconut coir pith in solid state fermentation) Muniswaran, P.K.A.; Selvakumar, P.; Charyulu, N.C.L.N.1994Item Production of novel cell-associated tannase from newly isolated Serratia ficaria DTC(2010) Belur, P.D.; Gopal, M.; Nirmala, K.R.; Nainegali, N.Five strains of tannic acid degrading bacteria were isolated and identified by phenotypic characterization. All the five isolates showed cell-associated activity, whereas only three showed extracellular activity. Serratia ficaria DTC, showing the highest cell-associated activity (0.29 U/l), was selected for further shake-flask studies. Tannase synthesis was growth associated and reached the peak in the late stationary phase of growth. Organic nitrogen sources enhanced the tannase production. Peak tannase production of 0.56 U/l was recorded in the medium having the initial pH of 6. The pH and temperature optima of the enzyme were found to be 8.9 and 35°C, respectively. This is the first report of cell-associated activity in the case of bacterial tannase. Cell-associated tannase of Serratia ficaria DTC could be industrially important from the perspective of its activity at broad temperature and pH ranges, and its unusually high activity at pH 8.9. © The Korean Society for Microbiology and Biotechnology.Item Optimization of culture medium for novel cell-associated tannase production from bacillus massiliensis using response surface methodology(2012) Belur, P.D.; Goud, R.; Goudar, D.C.Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A 2 3 factorial design augmented by 7 axial points (? = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, 30°C. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid. © The Korean Society for Microbiology and Biotexhnology.Item Production of propyl gallate in nonaqueous medium using cell-associated tannase of Bacillus massiliensis: Effect of various parameters and statistical optimization(2013) Aithal, M.; Belur, P.D.Enzymatic synthesis of propyl gallate in an organic solvent was studied using cell-associated tannase (E.C. 3.1.1.20) of Bacillus massiliensis. Lyophilized biomass showing tannase activity was used as a biocatalyst. The influence of buffer pH and strength, water activity, temperature, biocatalyst loading, gallic acid concentration, and 1-propanol concentration was studied by the one-factor-at-a-time method. Subsequently, response surface methodology was applied based on a central composite design to determine the effects of three independent variables (biocatalyst loading, gallic acid concentration, and 1-propanol concentration) and their mutual interactions. A total of 20 experiments were conducted, and a statistical model was developed, which predicted the maximum propyl gallate yield of 20.28 ?g/mL in the reaction mixture comprising 40.4 mg biocatalyst, 0.4 mM gallic acid, and 6.52 % (v/v) 1-propanol in 9.5 mL benzene at 30°C. The subsequent verification experiments established the validity of the model. Under optimal conditions, 25% conversion of gallic acid to propyl gallate was achieved on a molar basis. The absence of the need for enzyme purification and subsequent immobilization steps and good conversion efficiency makes this enzyme system an interesting one. Reports on the applications of bacterial whole cell systems for synthetic reactions in organic solvents are scarce, and perhaps this is the first report on bacterial cell-associated tannase-mediated esterification in a nonaqueous medium. © 2013 International Union of Biochemistry and Molecular Biology, Inc.Item Production of Naringinase by a new soil isolate of Serratia Sp.: Effect of different carbon and nitrogen sources(2013) Pavithra, M.; Belur, P.D.; Saidutta, M.B.Four strains of Naringin degrading bacteria were isolated and tested for naringinase activity. All the four isolates showed extracellular naringinase activity. The one which showed consistently good activity in three different media was selected (2 U/L) and was identified by phenotypic characterization as Serratia Sp. In shake-flask trials, effect of various carbon and nitrogen sources was studied. Among all the carbon sources, glucose enhanced the naringinase production. Peptone supplemented with ammonium nitrate was found to be favourable. Maximum of 9.2 U/L naringinase activity was achieved in the medium comprising naringin, glucose, peptone, ammonium nitrate and salts.Item Partial purification and characterization of L-asparaginase from an endophytic Talaromyces pinophilus isolated from the rhizomes of Curcuma amada(Elsevier, 2016) Krishnapura, P.R.; Belur, P.D.l-Asparaginase is a commercially significant enzyme. There exists a demand for a broad variety of microbial l-asparaginases with characteristics compatible with its different applications. Endophytic microorganisms, in particular are emerging as potential sources of l-asparaginases. The current work involves partial purification and characterization of l-asparaginase from Talaromyces pinophilus, an endophytic fungus isolated from the rhizomes of Curcuma amada. Maximum enzyme activity could be achieved at pH 8.0 and with temperature 28 °C. The enzyme Exhibits 95 % and 98% of its total activity at physiological pH and temperature, respectively. The enzyme activity is largely unhindered in the presence of metal ions such as Ca2+, Cu2+, Fe2+, Mg2+, Mn2+, Zn2+. Increase in the enzyme activity in the presence of thiol groups and reduction in the same upon addition of thiol blockers indicates the involvement of cysteine in the enzyme's catalytic activity. The enzyme is a heterodimer of 62 kDa and 39 kDa. The enzyme has a Km of 6.4 mM, its turnover number towards l-asparagine is 286.3 s-1. The enzyme has 16% glutaminase activity; its Km towards glutamine is 13.3 mM and turnover number is 54.6 s-1. Our results highlight that l-asparaginase from endophytic Talaromyces pinophilus could be considered as potential candidate for clinical and industrial trials, owing to its efficiency and biochemical properties. To the best of our knowledge, this is the first report on partial purification and characterization of L-asparaginase from an endophyte. © 2015 Elsevier B.V. All rights reserved.Item Novel Indole-Quinazolinone Based Amides as Cytotoxic Agents(HeteroCorporation support@jhetchem.com, 2016) Gokhale, N.; Panathur, N.; Udayakumar, D.; Nayak, P.G.; Pai, K.S.R.Indole-quinazolinone hybrids with active amides were synthesized, characterized, and assessed for their cytotoxicity. Two molecules displayed substantial activity in sulphorhodamine B assay method. © 2015 HeteroCorporation.Item Pathway identification, enzyme activity and kinetic study for the biodegradation of phenol by Nocardia hydrocarbonoxydans NCIM 2386(Taylor and Francis Inc. 325 Chestnut St, Suite 800 Philadelphia PA 19106, 2016) Shetty, G.R.; Shetty K, K.V.Nocardia hydrocarbonoxydans NCIM 2386 (Nhy) can grow using phenol as a sole carbon source and has a strong ability to degrade phenol. The paper presents the main metabolism pathways and mechanism of phenol degradation by Nhy. Phenol was found to be degraded via meta cleavage of catechol by the action of enzyme catechol 2,3-dioxygenase. The enzyme was found to be both extracellular and cell bound. The cell bound and extracellular enzymes actively degraded phenol even in the absence of the organism. The rate of phenol degradation by extracellular enzymes as sole enzymatic process (in the absence of cells) was found to be almost similar to that with the whole cells, indicating the prominence of extracellular enzymes. Michaelis–Menten model was found to fit the degradation rate kinetics of total phenol for total phenol concentrations of less than 100 mg L?1and also the degradation rate kinetics of catechol at catechol concentrations of less than 80 mg L?1during the exponential growth phase of the organism. Michaelis– Menten model was found to fit the kinetics of catechol formation rate which is also equal to the actual rate of phenol degradation to catechol. Both phenol and catechol were found to be substrate inhibitory. © 2015 Balaban Desalination Publications. All rights reserved.Item Metabolomic profiling of doxycycline treatment in chronic obstructive pulmonary disease(Elsevier B.V., 2017) Singh, B.; Jana, S.K.; Ghosh, N.; Das, S.K.; Joshi, M.; Bhattacharyya, P.; Chaudhury, K.Serum metabolic profiling can identify the metabolites responsible for discrimination between doxycycline treated and untreated chronic obstructive pulmonary disease (COPD) and explain the possible effect of doxycycline in improving the disease conditions. 1H nuclear magnetic resonance (NMR)-based metabolomics was used to obtain serum metabolic profiles of 60 add-on doxycycline treated COPD patients and 40 patients receiving standard therapy. The acquired data were analyzed using multivariate principal component analysis (PCA), partial least-squares-discriminant analysis (PLS-DA), and orthogonal projection to latent structure with discriminant analysis (OPLS-DA). A clear metabolic differentiation was apparent between the pre and post doxycycline treated group. The distinguishing metabolites lactate and fatty acids were significantly down-regulated and formate, citrate, imidazole and L-arginine upregulated. Lactate and folate are further validated biochemically. Metabolic changes, such as decreased lactate level, inhibited arginase activity and lowered fatty acid level observed in COPD patients in response to add-on doxycycline treatment, reflect the anti-inflammatory action of the drug. Doxycycline as a possible therapeutic option for COPD seems promising. © 2016 Elsevier B.V.Item Anionic surfactant based reverse micellar extraction of l-asparaginase synthesized by Azotobacter vinelandii(Springer Verlag, 2017) Murugesan, S.; Iyyaswami, R.; Kumar, S.V.; Surendran, A.Abstract: l-Asparaginase synthesized by Azotobacter vinelandii via submerged fermentation in the presence of sucrose was successfully extracted using Reverse micellar extraction. Single step enzyme purification process was developed by varying the process variables which resulted in maximum specificity and extraction of l-asparaginase. The effect of different variables, including broth pH, addition of alcohol during the forward extraction and pH of the fresh stripping aqueous phase, addition of alcohol and electrolyte during backward extraction process were studied. Lower concentration of butanol resulted in maximum activity of the enzyme during forward extraction while enzyme activity was found to increase further with the addition of higher concentrations of ammonium sulphate during backward extraction. Chromatographic analysis of l-asparaginase peak at ~7.65 min was intense for the back extracted sample confirming the maximum purity of l-asparaginase obtained. Purity of l-asparaginase was increased to about 379.68 fold. Graphical abstract: [Figure not available: see fulltext.]. © 2017, Springer-Verlag Berlin Heidelberg.