Skip to main content
×
Home
    • Aa
    • Aa

Isolation of lactoperoxidase using different cation exchange resins by batch and column procedures

  • Leonard WT Fweja (a1), Michael J Lewis (a1) and Alistair S Grandison (a1)
Abstract

Lactoperoxidase (LP) was isolated from whey protein by cation-exchange using Carboxymethyl resin (CM-25C) and Sulphopropyl Toyopearl resin (SP-650C). Both batch and column procedures were employed and the adsorption capacities and extraction efficiencies were compared. The resin bed volume to whey volume ratios were 0·96:1·0 for CM-25C and ⩽0·64:1·0 for SP-650 indicating higher adsorption capacity of SP-650 compared with CM-25C. The effluent LP activity depended on both the enzyme activity in the whey and the amount of whey loaded on the column within the saturation limits of the resin. The percentage recovery was high below the saturation point and fell off rapidly with over-saturation. While effective recovery was achieved with column extraction procedures, the recovery was poor in batch procedures. The whey-resin contact time had little impact on the enzyme adsorption. SDS PAGE and HPLC analyses were also carried out, the purity was examined and the proteins characterised in terms of molecular weights. Reversed phase HPLC provided clear distinction of the LP and lactoferrin (LF) peaks. The enzyme purity was higher in column effluents compared with batch effluents, judged on the basis of the clarity of the gel bands and the resolved peaks in HPLC chromatograms.

Copyright
Corresponding author
*For correspondence; e-mail: a.s.grandison@reading.ac.uk
References
Hide All
Casal E, Montilla A, Moreno FJ, Olano A & Corzo N 2006 Use of chitosan for selective removal of beta lactoglobulin from whey. Journal of Dairy Science 89 13841389
Chiu CK & Etzel MR 1997 Fractionation of lactoperoxidase and lactoferrin from bovine whey using a cation exchange membrane. Journal of Food Science 62 996–1000
De Wit JN & Van Hooydonk ACM 1996 Structure, functions and applications of lactoperoxidase in natural antimicrobial systems. Netherlands Milk and Dairy Journal 50 227244
Foley AA & Bates GW 1987 The Purification of lactoferrin from human whey by batch extraction. Analytical Biochemistry 162 296300
Fonteh FA 2001 Role of the lactoperoxidase system in raw milk preservation, PhD Thesis, University of Reading
Fonteh FA, Grandison AS & Lewis MJ 2002 Variations of lactoperoxidase activity and thiocyanate content in cows' and goats' milk throughout lactation. Journal of Dairy Research 69 401409
Goodall S, Grandison AS, Jauregi P & Price J 2008 Selective separation of the major whey proteins using ion exchange membranes. Journal of Dairy Science 91 110
Hernández MCM, Markwijik V & Vreeman HJ 1990 Isolation and properties of lactoperoxidase from bovine milk. Netherlands Milk and Dairy Journal 44 213231
Morrison M, Hamilton HB & Stotz E 1957 The isolation and purification of lactoperoxidase by ion exchange chromatography. Journal of Biological Chemistry 228 767776
Paul KGH, Ohlsson PI & Henriksson A 1980 The Isolation and some ligandic properties of lactoperoxidase. FEBS Letters 110 200204
Polis BD & Shmukler HN 1953 Crystalline lactoperoxidase. Isolation by displacement chromatography. Journal of Biological Chemistry 201 475500
Rombatus WA, Schroeder WA & Morrison M 1967 Bovine lactoperoxidase. Partial characterisation of the further purified protein. Biochemistry 6 29652977
Uchida T, Dosako S, Sato K & Kawakami H 2003 Sequential separation of lactoferrin, lactoperoxidase and secretory components by sulfate-linked ion exchange chromatography. Milchwissenschaft 58 482486
Yoshida S 1988 Isolation of some minor proteins distributed in acid whey from approximately 100,000 to 250,000 Daltons of particle size. Journal of Dairy Science 71 19
Yoshida S & Ye-Xiuyun 1991a Isolation of lactoperoxidase and lactoferrin from bovine milk rennet whey and acid whey by sulphopropyl cation-exchange chromatography. Netherlands Milk and Dairy Journal 45 273280
Yoshida S & Ye-Xiuyun 1991b Isolation of lactoperoxidase and lactoferrin from bovine milk acid whey by carboxymethyl cation exchange chromatography. Journal of Dairy Science 74 14391444
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Journal of Dairy Research
  • ISSN: 0022-0299
  • EISSN: 1469-7629
  • URL: /core/journals/journal-of-dairy-research
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords:

Metrics

Full text views

Total number of HTML views: 3
Total number of PDF views: 17 *
Loading metrics...

Abstract views

Total abstract views: 102 *
Loading metrics...

* Views captured on Cambridge Core between September 2016 - 20th October 2017. This data will be updated every 24 hours.