Lactoperoxidase Immobilized Onto Various Beads For Producing Natural Preservatives Solution

Ahmad Ni'Matullah Al-Baarri,S Hayakawa,M Ogawa,T Visalsok

doi:10.17728/jatp.52

Abstract

The natural preservative agent of hypothiocyanite (OSCN – ) has been succeded to determine using immobilized LPO onto various types of beads or matrixes: SP Sepharose Fast Flow (SP-FF), SP Sepharose Big Beads (SP-BB), CNBr activated SP Sepharose (SP-CNBR). Lactoperoxidase (LPO) was obtained from bovine milk. The immobilization method was performed using ion exhange method and adsorption. Immobilized LPO onto matrixes has been stored for 7 days at 4˚C and 10˚C. Data of the production of OSCN – were collected before and after immobilized LPO storage. Data shows no remarkable effect of the production of OSCN – using immobilized LPO stored at 4˚C, however, the remarkable reduction has been detected in the production of OSCN – using immobilized LPO that was stored at 10˚C. As a conclusion, the 7 days storage does not change the capability of immobilized LPO for producing OSCN if the immobilized LPO was stored at 4˚C.

Highlights

LPO in the combination with SCN− and H2O2, known as LPOs, generates hypothiocyanite OSCN− that kills microorganisms by oxidizing sulphydryl (–SH) groups of microbe’s cell membrane proteins (Seifu et al, 2005, Touch et al, 2004, Singh et al, 2009)
Hypothiocyanite produced by LPOs has been shown to be effective against pathogenic bacteria (Borch et al, 1989) as well as non-‐pathogenic bacteria (Wolfson and Sumner, 1993, Fonteh et al, 2005, Wakabayashi et al, 2007)
Another is that immobilized enzymes are separated from the reaction products and, reusable (Zhou and Lim, 2009, Tzanov et al, 2002, Choi and Yiu, 2004). These features contribute to reduce the cost of the production of chemical substances using enzyme-‐catalyzed reactions (Hwang et al, 2004, Altun and Cetinus, 2007). Such an effective biocatalytic reaction using immobilization technology let us challenge the preparation of immobilized LPO

Summary

INTRODUCTION

LPO in the combination with SCN− and H2O2, known as LPOs, generates hypothiocyanite OSCN− that kills microorganisms by oxidizing sulphydryl (–SH) groups of microbe’s cell membrane proteins (Seifu et al, 2005, Touch et al, 2004, Singh et al, 2009). One is that an enzyme can acquire markedly high thermal and operational stability by virtue of the interaction with immobilization matrix (Altun and Cetinus, 2007, Mahmoud and Helmy, 2009, Iyer and Ananthanarayan, 2008). Another is that immobilized enzymes are separated from the reaction products and, reusable (Zhou and Lim, 2009, Tzanov et al, 2002, Choi and Yiu, 2004). The used immobilized LPO was stored at 4 ̊C and 10 ̊C for 7 days to explore the feasibility of production of OSCN−

MATERIALS AND METHODS

Immobilization procedure

RESULTS AND DISCUSSION

CONCLUSION