Lactoperoxidase purification from whey by using dye affinity chromatography

NICOLÁS URTASUN; MARÍA FERNANDA BAIELI; DANIELA BELÉN HIRSCH; MARÍA CAMILA MARTÍNEZ-CERÓN; OSVALDO CASCONE; FEDERICO JAVIER WOLMAN

FOOD AND BIOPRODUCTS PROCESSING

INST CHEMICAL ENGINEERS

Año: 2017 vol. 103 p. 58 - 65

0960-3085

tBovine lactoperoxidase is aglycoprotein present in milk, whey and colostrum, which mightbe used in dairy,cosmetic, pharmaceutical, veterinary and agricultural applications due toitsbroad antimicrobial activity. Here, we describe a novel process for bovinelactoperoxidasepurification by using dye affinity chromatography. Eighteentriazine dyes were immobilized on Sepharose 6B and screened for theirperfor-mance as possible ligands. Five of the dye-Sepharose matrices showedover 90% adsorptionof bovine lactoperoxidase directly from whey without anypretreatment using the batchmode, and were thus selected for further adsorptionand elution studies. The highest elu-tion degree was obtained using 20 mMacetate buffer, pH 5.0, 2 M NaCl, as the eluent for allthe matrices. Wheyprocessed using the Reactive Red 4-Sepharose matrix in batch modeshowed thehighest bovine lactoperoxidase purification yield (86.5 ± 3.8%),purification fac-tor (46.1 ± 1.1), and arelative purity higher than 80% according to SDS-PAGE gel densitometry.Wheyprocessed using packed-bed column mode showed lower yields and additionalwheypretreatments were needed for dynamic processing.The interaction betweenbovine lactoperoxidase and Reactive Red 4-Sepharose matrix wascharacterizedusing Langmuir isotherm model. The Kdvalue was 0.21± 0.03 mg/mL and theQmaxwas32.21 ± 1.24 mg/g. The results presented heresuggest the potential application ofthe Reactive Red 4-Sepharose matrix toone-step purification of bovine lactoperoxidase fromwhey.© 2017 Institution of ChemicalEngineers.