Faculty Publications
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Item Production and characterization of biosurfactant produced by a novel Pseudomonas sp. 2B(2012) Aparna, A.; Srinikethan, G.; Smitha, H.Biosurfactant-producing bacteria were isolated from terrestrial samples collected in areas contaminated with petroleum compounds. Isolates were screened for biosurfactant production using Cetyl Tri Ammonium Bromide (CTAB)-Methylene blue agar selection medium and the qualitative drop-collapse test. An efficient bacterial strain was selected based on rapid drop collapse activity and highest biosurfactant production. The biochemical characteristics and partial sequenced 16S rRNA gene of isolate, 2B, identified the bacterium as Pseudomonas sp. Five different low cost carbon substrates were evaluated for their effect on biosurfactant production. The maximum biosurfactant synthesis (4.97g/L) occurred at 96h when the cells were grown on modified PPGAS medium containing 1% (v/v) molasses at 30°C and 150rpm. The cell free broth containing the biosurfactant could reduce the surface tension to 30.14mN/m. The surface active compound showed emulsifying activity against a variety of hydrocarbons and achieved a maximum emulsion index of 84% for sunflower oil. Compositional analysis of the biosurfactant reveals that the extracted biosurfactant was a glycolipid type, which was composed of high percentages of lipid (~65%, w/w) and carbohydrate (~32%, w/w). Fourier transform infrared (FT-IR) spectrum of extracted biosurfactant indicates the presence of carboxyl, hydroxyl and methoxyl functional groups. The mass spectra (MS) shows that dirhamnolipid (l-rhamnopyranosyl-l-rhamnopyranosyl-3-hydroxydecanoyl-3-hydroxydecanoate, Rha-Rha-C 10-C 10) was detected in abundance with the predominant congener monorhamnolipid (l-rhamnopyranosyl-?-hydroxydecanoyl-?-hydroxydecanoate, Rha-C 10-C 10). The crude oil recovery studies using the biosurfactant produced by Pseudomonas sp. 2B suggested its potential application in microbial enhanced oil recovery and bioremediation. © 2012 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.Item Low frequency sonic waves assisted cloud point extraction of polyhydroxyalkanoate from Cupriavidus necator(Elsevier B.V., 2017) Murugesan, S.; Iyyaswami, R.Low frequency sonic waves, less than 10 kHz were introduced to assist cloud point extraction of polyhydroxyalkanoate from Cupriavidus necator present within the crude broth. Process parameters including surfactant system variables and sonication parameters were studied for their effect on extraction efficiency. Introduction of low frequency sonic waves assists in the dissolution of microbial cell wall by the surfactant micelles and release of cellular content, polyhydroxyalkanoate granules released were encapsulated by the micelle core which was confirmed by crotonic acid assay. In addition, sonic waves resulted in the separation of homogeneous surfactant and broth mixture into two distinct phases, top aqueous phase and polyhydroxyalkanoate enriched bottom surfactant rich phase. Mixed surfactant systems showed higher extraction efficiency compared to that of individual Triton X-100 concentrations, owing to increase in the hydrophobicity of the micellar core and its interaction with polyhydroxyalkanoate. Addition of salts to the mixed surfactant system induces screening of charged surfactant head groups and reduces inter-micellar repulsion, presence of ammonium ions lead to electrostatic repulsion and weaker cation sodium enhances the formation of micellar network. Addition of polyethylene glycol 8000 resulted in increasing interaction with the surfactant tails of the micelle core there by reducing the purity of polyhydroxyalkanoate. © 2017 Elsevier B.V.Item Reverse micellar extraction of lactoferrin from its synthetic solution using CTAB/n-heptanol system(Springer India sanjiv.goswami@springer.co.in, 2017) Pawar, S.S.; Iyyaswami, R.; Belur, P.D.The partitioning of Lactoferrin (LF) into the reverse micellar phase formed by a cationic surfactant, cetyltrimethylammonium bromide (CTAB) in n-heptanol from the synthetic solution of LF was studied. The solubilization behaviour of LF into the reverse micellar phase and back extraction using a fresh stripping phase were improved by studying the effect of processing parameters, including surfactant concentration, solution pH, electrolyte salt concentration and addition of alcohol as co-solvent. Forward extraction of 100% was achieved at CTAB concentration of 50 mM in n-heptanol solvent, pH of 10 and 1 M NaCl. The electrostatic force and hydrophobic interaction have major influence on LF extraction during forward and back extraction respectively. The size of the reverse micelles and their corresponding water content were measured at different operating conditions to assess their role on the LF extraction. The present reverse micellar system has potential to solubilise almost all the LF into the reverse micelles during forward extraction and could able to allow back extraction from the reverse micellar phase with addition of small amount of co-solvent. © 2017, Association of Food Scientists & Technologists (India).Item Analysis of ionic and nonionic surfactants blends used for the reverse micellar extraction of Lactoperoxidase from whey(John Wiley and Sons Ltd, 2021) Karanth, S.; Iyyaswami, I.Bovine Lactoperoxidase (LP), a minor whey protein, is used as an antimicrobial in cosmetic, food, and pharmaceutical preparations. Industries are in pursuit of reliable, cheap, and scalable purification methods as the conventional techniques for LP purification like chromatography and membrane separation suffer from several drawbacks. The present work investigates the selective reverse micellar extraction of LP using the reverse micellar system formed by mixing food grade nonionic (Tween, Span, and Triton series) and ionic (AOT) surfactant blends. The analysis of LP extraction efficiency was performed by varying the concentration of nonionic surfactants with a constant AOT concentration of 100 mM and the initial pH of the system. Complete LP solubilization was achieved with reverse micelles formed by 100 mM AOT and 20 mM Tween 80 at pH 8. It was found that the extraction efficiency was dependent on the chain length or the number of ethylene oxide units in the Triton surfactant tail and the carbon–carbon double bond in Tween 80 tail, that is, on oleic acid. Span series however showed poor extraction in the organic phase substantiating the lesser water content. The forward extracted LP was successfully back-extracted into a fresh aqueous phase containing 1 M KCl at pH 10.5. The aqueous phase (whey) from the forward transfer can be further used to fractionate other whey proteins. © 2020 Curtin University and John Wiley & Sons, Ltd.Item Mixed Surfactant-Based Reverse Micellar Extraction Studies of Bovine Lactoperoxidase(John Wiley and Sons Inc, 2021) Karanth, S.; Iyyaswami, I.The suitability of reverse micellar extraction for recovery of bovine lactoperoxidase (LP) from aqueous solution was evaluated using systems formed by ionic and nonionic surfactant mixtures. The influence of ionic surfactant concentration, organic solvent, and pH on the extraction of LP into the reverse micellar phase was studied. The Tween® series surfactants with Aerosol-OT (bis-(2-ethylhexyl) sulfosuccinate) showed better extraction of LP in the reverse micelles (RM) compared to the Triton® and Span® series of surfactants. Complete extraction of LP from an aqueous phase of initial concentration 25 mg L?1 occurred with the RM formed by 90 mM Aerosol-OT/8 mM Tween® 80 in isooctane. The optimal pH, ionic strength, and positively charged ionic surfactant concentration for back extraction were also studied and a maximum of 95.5% back extraction efficiency and 66% LP activity recovery was obtained for a pH of 10.5,1 M KCl and 60 mM cetyltrimethylammonium bromide system. © 2021 AOCSItem Lactoperoxidase partitioning from whey using the reverse micelles of non-ionic/ionic mixed surfactants: Improvement of back extraction(John Wiley and Sons Inc, 2022) Karanth, S.; Iyyaswami, R.The reverse micellar system formed with a mixture of ionic and non-ionic surfactants, AOT (Sodium bis[2-ethyl hexyl] sulfosuccinate)/Tween 80 in isooctane, was studied for the selective extraction of Bovine Lactoperoxidase (LP) from acid whey. The effect of pH and ionic strength of acid whey and surfactant concentrations were studied and a maximum of 86% LP was extracted from acid whey at pH 9.5 with the addition of 0.2 M KCl to the reverse micelles formed with AOT (115 mM)/Tween 80 (23 mM). The back extraction of LP was studied at different aqueous stripping phase pH, ionic strength and concentration of counter-ionic surfactant Cetyltrimethylammonium bromide (CTAB). The back extraction of 112% with 80% LP recovery was achieved when the stripping phase pH was 10.5 with 1.5 M KCl and 60 mM CTAB. The antimicrobial activity of the extracted LP showed reduction in colony-forming units of S. aureus. Novelty impact statement: The reverse micelles formed with AOT/Tween 80 surfactant mixture minimize the pH-dependent denaturation of LP and widen the pH window (7.5–9.5) for LP extraction. The back extraction of LP from the reverse micellar phase to aqueous stripping phase was improved by the addition of CTAB as counter-ionic surfactant. A purification fold of 11.26 achieved with minimal loss in activity of LP by retaining the native Antimicrobial characteristic. © 2022 Wiley Periodicals LLC.Item Biosurfactant Based Reverse Micellar Extraction of Lactoperoxidase from Whey: Exploitation of Rhamnolipid Characteristics for Back Extraction(Taylor and Francis Ltd., 2023) Karanth, S.; Iyyaswami, R.; Raj, N.T.Biosurfactant-based reverse micellar extraction of Lactoperoxidase (LP) was studied using Rhamnolipid (RL) as a biosurfactant. Different solvents were considered to select a suitable organic phase for forming reverse micelles (RM) to varying concentrations of RL for the extraction of LP from its synthetic aqueous solution. The effect of addition of nonionic surfactant as lipophilic linker, whey pH, and ionic strength of the whey was studied to improve the forward extraction of LP from acid whey. About 96.65% LP was extracted to the RM phase during forward extraction. Further, a new back extraction strategy was developed by harnessing the biosurfactant properties. The pH-specific protonation–deprotonation characteristic of the RL headgroups was exploited to overcome the back extraction of LP, which is the rate-limiting step. The back extraction in citrate buffer at pH 5 using 0.75 M KCl resulted in 85.71% active LP recovery with 8.4-fold purification. The effect of the extraction process on the antimicrobial activity of LP was further examined with S. aureus, and the multiplication of the organism was almost arrested even after 24 hr at 9°C. © 2023 Taylor & Francis Group, LLC.Item Experimental investigation for treating ibuprofen and triclosan by biosurfactant from domestic wastewater(Academic Press, 2023) Jayalatha, N.A.; Devatha, C.P.The presence of emerging pollutants of pharmaceutical products and personal care products (PPCPs) in the aquatic environment overspreads the threat on living beings. Bioremediation is a promising option for treating wastewater. In the present study, an experimental investigation was carried out to produce a biosurfactant by Pseudomonas aeruginosa (MTCC 1688) for the removal of Ibuprofen (IBU) and Triclosan (TCS) from domestic wastewater. It was performed in three stages. Firstly, the production and optimization of biosurfactant was carried out to arrive at the best combination of crude sunflower oil, sucrose and ammonium bicarbonate (10%: 5.5 g/L: 1 g/L) to yield effective biosurfactant production (crude biosurfactant) and further extended to achieve critical micelle concentration (CMC) formation by dilution (biosurfactant at 10.5%). The stability of the biosurfactant was also confirmed. Biosurfactant showed a reduction in the surface tension to 41 mN/m with a yield concentration of 11.2 g/L. Secondly, its effectiveness was evaluated for the removal of IBU and TCS from the domestic wastewater collected during the dry and rainy seasons. Complete removal of IBU was achieved at 36 h & 6 h and TCS at 6 h & 1 h by crude biosurfactant and biosurfactant at CMC formation for the dry season sample. IBU removal was achieved in 2 h by both crude and biosurfactant at CMC and no TCS was detected in the rainy season sample. Thirdly, biotransformation intermediates of IBU and TCS formed during the application of the biosurfactant and degradation pathways are proposed based on the Liquid Chromatography-Mass Spectrometry (LC-MS) and it indicates that there is no formation of toxic by-products. Based on the results, it is evident that biosurfactant at CMC has performed better for the removal of IBU and TCS than crude biosurfactants without any formation of toxic intermediates. Hence, this study proved to be an eco-friendly, cost-effective and sustainable treatment option for domestic wastewater treatment. © 2022 Elsevier LtdItem Extraction of chrysin from propolis and its selective encapsulation in synthetic/natural surfactant-based micelles(Taylor and Francis Ltd., 2024) Sivanesan, M.; Krishnapura, P.R.; Iyyaswami, R.; Parappa, K.; Belur, P.D.The encapsulation characteristics of chrysin (important flavonoid with potential food, pharmaceutical, and biomedical applications) was studied with nonionic surfactants Triton X-114 (TX) and Quillaja Saponin (QS), individually. The factors influencing the encapsulation efficiency (EE) of standard chrysin that is surfactant concentration, pH, NaCl concentration, and chrysin concentration were analyzed. The maximum EE of standard chrysin was found to be 98.23 ± 1.63% with TX micelles and 83 ± 2.31% with QS micelles under the following conditions: 0.02 mg/mL standard chrysin, 5% NaCl, pH 7, and 4% w/w TX 6% w/w QS. Selective extraction of chrysin from propolis was tried using three extraction techniques namely Maceration, Microwave-assisted Extraction (MAE), and Maceration with Microwave-assisted Extraction (MMAE). MAE, which gave a chrysin yield of 3 mg/g, was deemed the most suitable method for chrysin extraction from propolis. This MAE crude extract was subjected to encapsulation under the conditions previously optimized for standard chrysin. Specific encapsulation of chrysin from the propolis crude extract was achieved, with an EE of 92 ± 0.86% with TX and 84.97 ± 1.34% with QS. The encapsulated chrysin was characterized using particle size analysis and antioxidant activity. TX system was found to be the most suitable for the encapsulation, as it was able to selectively encapsulate chrysin from propolis, despite the presence of other interfering flavonoids in the crude extract. The microwave-assisted extraction combined with surfactant-based micellar encapsulation can be said to be an effective process for the extraction and encapsulation of chrysin from propolis. © 2023 Taylor & Francis Group, LLC.