Position: Home > Articles > Isolation, Purification and Characterization of Tyrosinase from Ipomoea aquatica Forsk. Leaves
FOOD SCIENCE
2015,36
(15)
167-172
蕹菜叶酪氨酸酶的分离纯化与部分性质
作 者：
孙才云;方玲;万骥;傅婷;王丹;唐云明
单 位：
西南大学生命科学学院淡水鱼类资源与生殖发育教育部重点实验室三峡库区生态环境教育部重点实验室
关键词：
蕹菜叶;酪氨酸酶;分离纯化;性质
摘 要：
蕹菜叶经匀浆、缓冲液提取、硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、Superdex-200凝胶过滤层析,获得电泳纯的酪氨酸酶。该酶活力达到114.53 U/mg,酶活力回收率为11.33%,纯化倍数为99.59。全酶分子质量为77.60 k D,亚基分子质量为38.70 k D;最适温度为45℃,最适pH值为7.5,该酶在25~55℃及pH 6.0~8.0的范围内有较好的稳定性;在最适条件下测得其Km值为10.05 mmol/L;甲醇、乙醇、异丙醇及柠檬酸、抗坏血酸、Ca2+和Pb2+对该酶有抑制作用,Mn2+、Zn2+和Co2+对该酶具有一定的激活作用,尿素和十二烷基硫酸钠(sodium dodecyl sulfonate,SDS)对该酶活性影响不大,Li+、K+对该酶活性基本没有影响。
译 名：
Isolation, Purification and Characterization of Tyrosinase from Ipomoea aquatica Forsk. Leaves
作 者：
SUN Caiyun;FANG Ling;WAN Ji;FU Ting;WANG Dan;TANG Yunming;Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, School of Life Science, Southwest University;
关键词：
Ipomoea aquatica Forsk.leaves;;tyrosinase;;isolation and purification;;characterization
摘 要：
Electrophoresis-purity tyrosinase(TYR) from Ipomoea Aquatica Forsk leaves was obtained through homogenization, buffer solution extraction, ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Superdex-200 gel filtration chromatography. The specific activity of purified TYR was 114.53 U/mg, with a recovery of 11.33% and a purification factor of 99.59. The molecular weights of TYR and its subunit were 77.60 k D and 38.70 k D, respectively. TYR was relatively stable in the range of 25–55 ℃ and pH 6.0–8.0. Its optimum temperature and p H were 45 ℃ and 7.5, respectively. Furthermore, its Km was 10.05 mmol/L under the optimum conditions. The activity of TYR could be inhibited by methanol, ethanol, isopropanol and citric acid and ascorbic acid, as well as some metal ions such as Ca2+ and Pb2+. It could be activated by Mn2+, Zn2+ and Co2+; however, urea and SDS had little effect on its activity, and Li+ and K+ has no effect on its activity.
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