Faculty Publications
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Item A new strategy to refine crude Indian sardine oil(Japan Oil Chemists Society yukagaku@jocs-office.or.jp, 2017) Charanyaa, S.; Belur, B.D.; Iyyaswami, R.Current work aims to develop a refining process for removing phospholipids, free fatty acids (FFA), and metal ions without affecting n-3 polyunsaturated fatty acid (n-3 PUFA) esters present in the crude Indian sardine oil. Sardine oil was subjected to degumming with various acids (orthophosphoric acid, acetic acid, and lactic acid), conventional and membrane assisted deacidification using various solvents (methanol, ethanol, propanol and butanol) and bleaching with bleaching agents (GAC, activated earth and bentonite) and all the process parameters were further optimized. Degumming with 5%(w/w) ortho phosphoric acid, two stage solvent extraction with methanol at 1:1 (w/w) in each stage and bleaching with 3% (w/w) activated charcoal loading, at 80ºC for 10 minutes resulted in the reduction of phospholipid content to 5.66 ppm from 612.66 ppm, FFA to 0.56% from 5.64% with the complete removal of iron and mercury. Under these conditions, the obtained bleached oil showed an enhancement of n-3 PUFA from 16.39% (11.19 Eicosapentaenoic acid (EPA) + 5.20 Docosahexaenoic acid (DHA)) to 17.91% (11.81 EPA + 6.1 DHA). Replacing conventional solvent extraction with membrane deacidification using microporous, hydrophobic polytetrafluoroethylene membrane (PTFE), resulted in a lesser solvent residue (0.25% (w/w)) in the deacidified oil. In view of lack of reports on refining of n-3 PUFA rich marine oils without concomitant loss of n-3 PUFA, this report is significant. © 2017 by Japan Oil Chemists’ Society.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 Lipase mediated synthesis of rutin fatty ester: Study of its process parameters and solvent polarity(Elsevier Ltd, 2017) Chandrasekar, C.; Belur, P.D.; Iyyaswami, R.Lipophilization of antioxidants is recognized as an effective strategy to enhance solubility and thus effectiveness in lipid based food. In this study, an effort was made to optimize rutin fatty ester synthesis in two different solvent systems to understand the influence of reaction system hydrophobicity on the optimum conditions using immobilised Candida antartica lipase. Under unoptimized conditions, 52.14% and 13.02% conversion was achieved in acetone and tert-butanol solvent systems, respectively. Among all the process parameters, water activity of the system was found to show highest influence on the conversion in each reaction system. In the presence of molecular sieves, the ester production increased to 62.9% in tert-butanol system, unlike acetone system. Under optimal conditions, conversion increased to 60.74% and 65.73% in acetone and tert-butanol system, respectively. This study shows, maintaining optimal water activity is crucial in reaction systems having polar solvents compared to more non-polar solvents. © 2017 Elsevier LtdItem Enhancement of n-3 polyunsaturated fatty acid glycerides in Sardine oil by a bioimprinted cross-linked Candida rugosa lipase(Elsevier Inc. usjcs@elsevier.com, 2018) Sampath, C.; Belur, P.D.; Iyyaswami, R.Considering the advantages of bioimprinting and carrier free immobilization, cross-linked enzyme aggregates (CLEA) were prepared by using bioimprinted Candida rugosa lipase (CRL) with Bovine serum albumin (BSA), Polyethyleneimine and glutaraldehyde. Effect of various factors such as CRL-Oleic acid ratio, CRL-BSA ratio, CRL- Polyethyleneimine ratio, glutaraldehyde loading, cross-linking time etc., on lipase activity recovery and aggregate yield were studied and optimized. This immobilized lipase (CRL-CLEA) was used for the selective hydrolysis of ester linkages of non-PUFA glycerides, with an aim to concentrate EPA and DHA glycerides in the Sardine oil. Imprinting with oleic acid in the presence of ethanol and Tween 60, and further immobilization with co-aggregates and cross-linking agent showed 10.4 times higher degree of hydrolysis compared to free enzyme. As result, 2.83-fold increase of n-3 PUFA content in deacidified oil was obtained by using CRL-CLEA. The resultant oil had negligible di- and triglycerides content, proving higher efficiency in hydrolysing ester bonds of fatty acids, other than n-3 PUFA. Reusability studies showed CRL-CLEA could be reused up to 5 runs without a substantial reduction in its performance. Improvement in degree of hydrolysis, thermostability, efficiency of hydrolysis and reusability were achieved due to bioimprinting and subsequent immobilization of CRL in the form of CLEA. © 2017 Elsevier Inc.Item Aqueous two phase partitioning of Pisum sativum lectin in PEG/citrate salt system(Taylor and Francis Inc. 325 Chestnut St, Suite 800 Philadelphia PA 19106, 2018) Bommenahalli Shashidhara, R.; Iyyaswami, R.Pisum sativum lectin (Psl) is a metalloprotein which is in the center of research interest because of its HIV-1 reverse transcriptase inhibitory activity and mitogenic activity. The application of this lectin in various fields demands the economically feasible and scalable purification strategy other than affinity chromatography. The suitability of aqueous two phase system (ATPS) composed of poly ethylene glycol (PEG) with different salts (sodium citrate, potassium citrate, and ammonium citrate) was evaluated for better partitioning of Psl. The significant factors such as molar mass and concentration of PEG, type and concentration of salts, the effect of tie line length (TLL), ionic strength, and pH were studied to select a suitable system for better partitioning of Psl. ATPS comprising of 18% PEG 6000, 16% sodium citrate, 1% NaCl at the operating condition of pH 8, 40.23% of TLL, and the volume ratio of 1.32 was found to be the best system which gave a maximum partition coefficient and yield of 14.5% and 98.66%, respectively. © 2018, © 2018 Informa UK Limited, trading as Taylor & Francis Group.Item Aqueous two phase based selective extraction of mannose/glucose specific lectin from Indian cultivar of Pisum sativum seed(Elsevier B.V., 2019) Rashmi, B.S.; Iyyaswami, R.Pisum sativum lectin (Psl) being a high-value protein has marked its application in the biomedical and therapeutic field. Aqueous two phase extraction (ATPE) was implemented as a selective partitioning technique for the partial purification of Psl from its seeds. PEG/citrate based biodegradable aqueous two phase system (ATPS) was screened and the factors such as the type and concentration of citrate salts, molar mass and concentration of polyethylene glycol (PEG), tie line length (TLL) and additive (NaCl) concentration, pH, crude load and volume ratio were studied for the selective partition of Psl. The Psl was successfully extracted to the top phase in the ATPS formed with 18% PEG 6000/16% sodium citrate at 41.01% TLL, 2% NaCl and pH of 7.5. A volume ratio of 0.76 and a crude load of 20% showed maximum activity yield of 122.12% with the purification factor of 16.26. The subunits of Psl namely ? and ? were identified with a molecular weight of 6 and 18 kDa respectively during the purity analysis using SDS PAGE and HPLC. © 2019 Elsevier B.V.Item Simultaneous partitioning of multiple bioactive compounds from Garcinia indica rinds in a three-liquid-phase extraction systems(Taylor and Francis Ltd., 2025) Shanbhag, C.C.; Salimuddin, R.M.H.; Iyyaswami, R.; Belur, P.D.Simultaneous extraction and purification of principal bioactive compounds, anthocyanins (ACNs), garcinol (GL), and hydroxycitric acid (HCA) from the rinds of Garcinia indica (Kokum) fruits in a single-step using Three Liquid Phase Systems (TLPS) were investigated. Among the various phase-forming components studied, TLPS formed by n-hexane-ethanol-(NH4)2SO4-water system was considered for partitioning GL into the n-hexane-rich top phase, ACNs into the ethanol-rich middle phase, and HCA into the aqueous salt-rich bottom phase. The present system was even able to separate carbohydrates into the bottom phase, which can be detrimental to the stability of ACNs. The effect of n-hexane, ethanol, and (NH4)2SO4 concentration on the partitioning behavior of biomolecules was analyzed. The TLPS composed of water-n-hexane-ethanol-(NH4)2SO4 could purify and extract 95.08% of ACNs, 95.33% of GL, and 67.98% of HCA in a single-step extraction process while the other extraction methods require multi-step extraction process to separate these three compounds. The effect of pH studies on the partitioning characteristics of biomolecules revealed that pH 4 is optimum and more efficient than the native pH of the system to achieve maximum yield of all the bioactive compounds. © 2025 Taylor & Francis Group, LLC.