Isolation of bovine lactoferrin, lactoperoxidase and enzymatically prepared lactoferricin from proteolytic digestion of bovine lactoferrin using adsorptive membrane chromatography.
A new downstream procedure for the isolation of bovine lactoferrin (bLf), lactoperoxidase and bovine lactoferricin (LfcinB) from sweet cheese whey was developed at the laboratory scale, based on membrane adsorber technology. The procedure was upscaled later on to an industrially relevant scale for the purificationof sweet whey concentrate with a recovery yield for lactoferrin of more than 90%. Based on these results the industrial process for 1 x 10(8) kg whey per year was projected. These high-value proteins were downstreamed by using cation-exchange membrane systems (Sartobind S, Sartorius, Göttingen, Germany). These strongly acidic membranes trap proteins in its anionic form. The dynamic loading capacity for both proteins as well as the optimal elution profiles with sodium chloride gradients were derived from laboratory experiments using membrane modules with 15-75 cm2 membrane material. Further investigations were performed with 1 m2 modules in a continuous process mode. The enzymatic preparation of LfcinB from bLf was performed by pepsin hydrolysis and the isolation of LfcinB was directly carried out from the enzymatic digest mixture. The identification of the proteins was performed with matrix-assisted laser desorption ionisation mass spectrometry (MALDI-MS). LfcinB and bLf were both tested afterwards in biological assays in order to show not only the efficiency of the downstreaming process in regard to product quantity but also to product quality (biological activity).
Plate
Kerstin
K
University of Hannover, Institute for Technical Chemistry, Callinstrasse 3, D-30167 Hannover, Germany.
Beutel
Sascha
S
Buchholz
Heinrich
H
Demmer
Wolfgang
W
Fischer-Frühholz
Stefan
S
Reif
Oscar
O
Ulber
Roland
R
Scheper
Thomas
T
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D007781:Lactoferrin / Q000302:isolation & purification*; D007784:Lactoperoxidase / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C082171:lactoferricin B; D007784:Lactoperoxidase; D007781:Lactoferrin