Partial purification and preparation of bovine lactoperoxidase and characterization of kinetic properties of its immobilized form incorporated into cross-linked alginate films

Abstract

Lactoperoxidase (LPS), purified directly from bovine rennet whey by Toyopearl-SP cation-exchange chromatography and lyophilized by using dextran as supporting material, maintained almost 70 and 60% of its activity after almost 2 and 5 months storage at À18°C, respectively. Incorporation of the prepared LPS into alginate films between 0.08 and 0.69 mg/cm 2 (516-4325 U/cm 2 ) caused the immobilization of most of the enzyme and gave films with LPS activity between 0.05 and 2.8 U/cm 2 , determined in the presence of 8 lM H 2 O 2 . Between 2 and 24 lM H 2 O 2 concentrations, a two-fold increase in H 2 O 2 concentration caused 1.5-2.5-fold increase in LPS activity of films incorporated with 0.24-0.28 mg/cm 2 (1200 U/cm 2 ) LPS. The Q 10 and E a of immobilized enzyme activity between 4 and 16°C were 1.69 and 34.6 kJ/mol, respectively. However, in the 16-30°C range, the temperature change had almost no effect on LPS activity of films. The optimal activity of immobilized LPS was observed at pH 6.0, but the enzyme maintained 30-85% of its activity between pH 3.0 and 7.0. The immobilized LPS also had a high stability between pH 4.0 and 6.0. The results of this study showed the good potential of LPSincorporated alginate films in forming a natural antimicrobial mechanism in different foods.

Antimicrobial Activity of Lactoperoxidase System Incorporated into Cross‐Linked Alginate Films

“…The enzyme was prepared according to the method of Mecitoğlu and Yemenicioğlu (2007). For this purpose, LPS active fractions were pooled and dialyzed (cut off: 12.000 MW) for 24 h at 4 °C.…”

“…Recently, in our laboratories, the LPS has also been incorporated into alginate films. The enzyme incorporated into these edible films bound and immobilized effectively onto films following cross‐linking and it shows appropriate stability and activity at a broad temperature and pH range (Mecitoğlu and Yemenicioğlu 2007). In this study, the antimicrobial activity of LPS incorporated into alginate films and its components has been tested on different bacteria including E. coli , Listeria innocua , and Pseudomonas fluorescens.…”

Single step purification of lactoperoxidase from whey involving reverse micelles‐assisted extraction and its comparison with reverse micellar extraction

“…Purification of LPO was the subject of study for many research groups using various purification techniques. Ammonium sulfate precipitation in combination with cation‐exchange chromatography14 and gel filtration chromatography11 resulted in 50–64% and 28% recovery of LPO, respectively. These conventional techniques involved multi‐step procedure.…”

Purification of lactoperoxidase from bovine whey and investigation of kinetic parameters

“…Ozdemir et al .,[8] Nandini and Rastogi[27] and Mecitoğlu and Yemenicioğlu[9] used different successive methodes for separation LPO from bovine milk. However, by simply removing step of globulins, we use only CM-cellulose ion-exchange chromatography which is cost effective and short time-consuming.…”

“…[8] Mecitoğlu and Yemenicioğlu using toyopearl-sp Cation-exchange chromatography purified the enzyme. [9] In addition, affinity chromatography was used for the purification of LPO. [10] All of the research on the LPO purification have indicated that purification of the LPO can be performed using very-time consuming and complicated methods.…”