食品科学2017，Vol.38Issue(24)：20-27,8.DOI:10.7506/spkx1002-6630-201724004
金银花过氧化物酶的三相分离纯化及酶学性质
Purification of Peroxidase from Lonicera japonica Thunb.by Three Phase Partitioning and Enzymatic Properties
摘要
Abstract
Three phase partitioning was used to extract and purify peroxidase(POD)from Lonicera japonica Thunb.The optimal conditions were obtained as follows: pH 5.60, ammonium sulfate concentration 39.49 g/100 mL, and ratio of crude extract to t-butanol (V/V), 1:1.38. Under these conditions, the activity recovery was 87.64% with a purification factor of 5.849, and the specific activity of purified peroxidase was 1 021.6 U/mg. By using three phase partitioning, 92% pigment was removed. Enzymatic properties revealed that the optimum temperature for this enzyme was 30 ℃ and the optimum pH was 5. The enzyme was stable in the temperature range of 10–40 ℃ and in the pH range of 4–7. In the presence of both guaiacol and H2O2, Michaelis constants (Km) and maximum reaction rates (vmax) of the enzyme were 8.12 mmol/L and 1.71 mmol/(min·L) for guaiacol, and 0.822 mmol/L and and 1.38 mmol/(min·L) for H2O2at a fixed concentration of the other substrate, respectively. The affinity of the enzyme to various substrates was in the decreasing order: pyrogallol >catechol > bimethoxy aniline > guaiacol. The peroxidase activity was activated by Ca2+, Cu2+and Zn2+but inhibited by Mg2+, Mn2+, citric acid, ascorbic acid, L-cysteine, sodium sulphite and sodium metabisulphite.关键词
金银花/过氧化物酶/三相分离法/酶学性质Key words
Lonicera japonica Thunb./peroxidase/three phase partitioning/enzymatic properties分类
轻工纺织引用本文复制引用
罗磊,董金龙,朱文学,薛依涵,关宁宁,张宽,姬青华..金银花过氧化物酶的三相分离纯化及酶学性质[J].食品科学,2017,38(24):20-27,8.基金项目
国家自然科学基金联合基金项目(U1304330) （U1304330）
