Purification and characterization of two isozymes of chlorophyllase from mature leaves of Chenopodium album
Purification and characterization of two isozymes of chlorophyllase from mature leaves of Chenopodium album
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DOI：
10.1093/oxfordjournals.pcp.a029267
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发表时间：
1997-09-01
影响因子：
4.9
通讯作者：
Takamiya, K
中科院分区：
文献类型：
--
作者：
Tsuchiya, T;Ohta, H;Takamiya, K
从藜属植物成熟叶片中分离纯化了叶绿素酶，并对其酶学性质进行了研究。通过疏水层析、ConA琼脂糖层析、肝素亲和层析、Mono-Q离子交换层析、凝胶过滤等步骤，从白蜡树丙酮粉中提取纯化Chlase。ConA琼脂糖亲和层析和凝胶过滤是纯化效果最好的步骤。在Mono Q层析上，该Chlase制剂分离出具有Chlase活性的两个主要组分和一个次要组分。两种主要的Chlase被纯化为均一。经SDS-PAGE测定，其相对分子质量分别为41.3 kDa和40.2 kDa。两种酶的最适pH和最适K值相近。它们的N末端氨基酸序列几乎完全相同，只是其中一个Chlase的第10个氨基酸残基缺失；数据库搜索没有检测到同源蛋白。
Chlorophyllase (Chlase) was purified from mature leaves of Chenopodium album, and its enzymatic properties were investigated. Chlase was extracted from acetone powder of C. album and purified by the following chromatographic procedures: hydrophobic chromatography, Con A Sepharose, Heparin affinity chromatography, Mono Q ion-exchange chromatography, and gel-filtration. Con A Sepharose affinity chromatography and gel-filtration were the most effective steps on the purification. On Mono Q chromatography, the Chlase preparation separated into two major and one minor fractions that exhibited Chlase activity. The two major Chlases were purified to homogeneity. Their molecular masses were estimated as 41.3 kDa and 40.2 kDa by SDS-PAGE. The optimum pH and K, values of these two Chlases were similar. Their N-terminal amino acid sequences were almost identical except for a deletion in the tenth amino acid residue in one of the Chlase; there was no homologous protein detected by database search.