HPLC QUANTIFICATION OF LACTOPEROXIDASE IN THERAPEUTIC DAIRY WASTE ENRICHED BY BUBBLESEPARATION
DOI:
https://doi.org/10.22159/ijap.2019v11i4.32058Keywords:
Foam fractionation, Sodium dodecyl sulphate, Enrichment, Lactoperoxidase, Medicinal proteins, RP-HPLCAbstract
Objective: The objective of this study was to enrich therapeutic proteins and remove pollutants from dairy wastewater for establishing foam fractionation as a lucrative unit operation.
Methods: Dairy wastewater collected from dairy industry was processed to fat-free dried protein waste mass diluted to 1-liter feed by distilled water in different concentrations and foam fractionated by sodium dodecyl sulphate (surfactant) to enriched proteins extract (foamate) in a foam fractionator. Foamate were analysed to quantify total proteins and lacto peroxidase respectively. The efficiency was evaluated by varying parameters like pH, initial waste and ionic concentrations, the waste-surfactant mass ratio of feed and flow rate of gas (N2) through feed solution by several experiments. Heat of desorption (λ) and mass transfer coefficient (K) were determined as indicators of adsorptive bubble separation to foam phase governed by Gibb’s equation of adsorption isotherm.
Results: The process was optimized at pH 5.5, initial feed concentration 500μg/ml, waste–surfactant mass ratio (1.5:1), gas flow rate (350 ml/min) and ionic concentration 0.1 gram-mole of NaCL per litre of feed with enrichment factor (49.09), percent recovery (98.18%) observed in foamate. One natural preservative specifically lactoperoxidase was quantified by RP-HPLC analysis as 0.49% (w/w) of total proteins at optimal condition. Heat of desorption(λ), mass transfer coefficient(K)were determined 3140cal/mol and 12.68* 10-9 mol/cal/cm2/s respectively at pH 8.5, initial feed concentration 500μg/ml and gas flow rate 350 ml/min.
Conclusion: The method may be a useful unit operation for recovery of biomolecules and removal of toxic pollutants from industrial wastewater for coming days.
Downloads
References
Durham RJ, Hourigan JA. Waste management and co-product recovery in dairy processing. In: Waldron K. editor. Handbook of waste management and co-product recovery in food processing. 1st ed. England: Woodland Publishers Ltd; 2007. p. 332-87.
Bulletin IDF (International Diary Federation); 2017. p. 489.
UN-OECD (Organization for Economic Cooperation and Development): Special Report; 2017.
Lemlich R. Adsorptive bubble separation techniques. 1st ed. Academic Press: New York: Elsevier; 1972. p. 159-73.
Yuki YO, Chou J. Foam fractionation process. McGraw Hill Publishers: London; 1979.
Datta PK, Ghosh A, Chakraborty P, Gangopadhyay A. Foam fractionation in the separation of pharmaceutical biomolecules: a promising unit operation for industrial process and waste control. J Fundamental Pharm Res 2015;3:33-41.
Backeh M, Leupoid G, Ekichi P, Parlar H. Rapid quantitative extraction of carnosic acid from rosemary (Rosemarinous officinalis) by isoelectric focused adsorptive bubble chromatography. J Agric Food Chem 2003b;51:1297-301.
Holmstorm B. Foam fractionation of streptokinase from crude culture filtrates. J Biotechnol Bioeng 1968;10:551-2.
Rubio J, Tessele F. Removal of heavy metal ions by an adsorptive particulate floatation. J Min Eng 1997;10:671-9.
Kim YS, Choi YS, Lee YL. Determination of zinc and lead by solvent sublation using ion-pair. Bull Korean Chemical Society 2001;22:821-6.
Qu YH. Recovery of surfactant SDS and Cd2+from permeate in MEUF using a continuous foam fractionator. J Hazard Mater 2008;155:32-8.
Durham RJ, Hourigan JA, Sleigh RW, Johnson RL. Whey fractionation: wheying up the consequences. Food Australia 1997;49:460-5.
Jam MS, Van Hooijdonk ACM. Occurrence, structure, biochemical properties and technological characteristics of lactoferrin. Br J Nutr 2000;84 Suppl 1:11–7.
Giansanti F, Panella G, Labolfe L, Antonini G. Lactoferrin from milk: nutraceutical and pharmacological properties. J Pharm 2016;9:1-15.
Kussendrager KD, Van Hooijdonk ACM. Lactoperoxidase: physicochemical properties, occurrence, mechanism of action and applications. Br J Nutr 2000;84 Suppl 1:19-25.
Michel MJ, Pouliot Y, Britten M, Noel I, Lebrun R. Study on the molecular mass changes of bovine k-casein glycomacropeptide and on its separation in ultra-filtration with and without application of electric fields. J Diary Sci 2002;85:Suppl 1.
Marshall KR. Industrial isolation of milk proteins: Whey proteins. In: Fox PF. editor. Development in diary chemistry, proteins. ed. London: Chap. 11. Appl Sci Publishers; 1982.
Noel J, Prokop A, Tanner RD. Foam fractionation of a dilute solution of bovine lactoferrin. J Appl Biochem Biotechnol 2002;98-100:395-402.
Saleh ZS, Hossain HdM. A study of the separation of proteins from multi-component mixtures by a semi-batch foaming process. Chemical Engineering and Processing; Elsevier Publishers Ltd; 2001;40:371-8.
Besagni G, Inzoli F. Influence of electrolyte concentration on hold up flow regime transition and local flow properties in a large scale bubble column. 33rd Italian Union of Thermo fluid–dynamics Heat Transfer Conference. J Physics: Conference Series 2015:655. DOI:10.1088/1742-6596/655/1/012039.
Ekichi P, Backleh SM, Palar H. High-efficiency enrichment of total and single whey proteins by pH controlled foam fractionation. Int J Food Sci Nutr 2005;56:223–9.
Suzuki A, Yasuhara HS, Maruyama H. Selective foam separation of binary proteins solution by SDS complexation method. J Colloidal Interface Sci 2002;253:402-8.
Fuda E, Jauregi P, Pyle DL. Recovery of lactoferrin and lactoperoxidase from sweet whey using colloidal gas aphrons (CGAs) generated from an anionic surfactant AOT. J Biotechnol Progress 2004;20:514-25.
Published
How to Cite
Issue
Section
