Cankaya M, Sişecioğlu M, Bariş O, Güllüce M, Ozdemir H
Department of Chemistry, Faculty of Sciences, Atatürk University, 25240, Erzurum, Turkey.
Prikl Biokhim Mikrobiol. 2010 Jan-Feb;46(1):64-8.
Bovine lactoperoxidase (LPO) was purified from skimmed milk using amberlite CG-50-H+ resin, CM sephadex C-50 ion-exchange chromatography, and sephadex G-100 gel filtration chromatography. Lactoperoxidase was purified 20.45-fold with a yield of 28.8%. Purity of enzyme checked by sodium dodecyl sulphate-polyacrylamide gel electrophoresis method and a single band was observed. Km was 0.25 mM at 20 degrees C, Vmax value was 7.95 micromol/ml min at 20 degrees C (pH 6.0). Antibacterial study was done by disk diffusion method of Kir-by-Bauer using Mueller-Hinton agar medium with slight modification. Bovine LPO showed high antibacterial activity in 100 mM thiocyanate-100 mM H2O2 medium for some bacteria (Brevibacillus centrosaurus, B. choshinensis, B. lyticum, Cedecea davisae, Chryseobacterium indoltheticum, Clavibacter michiganense pv. insidiosum, Kocuria erythromyxa, K. kristinae, K. rosea, K. varians, Paenibacillus validus, Pseudomonas syringae pv. populans, Ralstonia pickettii, Rhodococcus wratislaviensis, Serratia fonticola, Streptomyces violaceusniger, Vibrio cholerae-nonO1) respectively, and compared with well known antibacterial substances (levofloxacin, netilmicin). LPO system has inhibition effects on all type bacteria and concentration is really important such as LPO-100 mM thiocyanate-100 mM H2O2 system was proposed as an effective agent against many factors causing several diseases.
使用amberlite CG - 50 - H⁺树脂、CM sephadex C - 50离子交换色谱和sephadex G - 100凝胶过滤色谱从脱脂牛奶中纯化牛乳铁过氧化物酶（LPO）。乳铁过氧化物酶纯化了20.45倍，产率为28.8%。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳法检查酶的纯度，观察到单一条带。20℃时Km为0.25 mM，20℃（pH 6.0）时Vmax值为7.95微摩尔/毫升·分钟。采用Kirby - Bauer纸片扩散法并对穆勒 - 欣顿琼脂培养基进行轻微改良进行抗菌研究。牛LPO在100 mM硫氰酸盐 - 100 mM H₂O₂培养基中对某些细菌（中心短芽孢杆菌、朝鲜短芽孢杆菌、溶胞短芽孢杆菌、戴维斯西地西菌、吲哚金黄杆菌、密执安棒形杆菌致病亚种、红皮考克氏菌、克里斯蒂考克氏菌、玫瑰考克氏菌、变异考克氏菌、有效类芽孢杆菌、丁香假单胞菌丁香致病变种、皮氏罗尔斯通氏菌、弗罗茨瓦夫红球菌、丰蒂氏沙雷氏菌、紫黑链霉菌、霍乱弧菌非O1型）分别表现出高抗菌活性，并与知名抗菌物质（左氧氟沙星、奈替米星）进行比较。LPO系统对所有类型细菌都有抑制作用，浓度非常重要，例如LPO - 100 mM硫氰酸盐 - 100 mM H₂O₂系统被认为是对抗多种致病因素的有效制剂。
