Purification and characterization of acetone cyanohydrin lyase from Linum usitatissimum.
Purification and characterization of acetone cyanohydrin lyase from Linum usitatissimum.
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DOI：
10.1016/0003-9861(88)90634-0
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发表时间：
1988-06
影响因子：
3.9
通讯作者：
L. L. Xu-L.;B. Singh;E. Conn
中科院分区：
文献类型：
--
作者：
L. L. Xu-L.;B. Singh;E. Conn
羟基腈裂解酶（EC 4.1.2._）从亚麻（LinumusitatissimumL.）幼苗中分离纯化了一种能催化丙酮和2-丁酮氰醇解离的化合物。纯化过程包括用（NH_4）_2SO_4沉淀，色谱聚焦，和在DEAE-纤维素、羟基磷灰石、Sephacryl 200和Matrex Red A凝胶柱上层析，最终回收率为21%。136倍的纯化产生了一个明显均匀的制剂，与从Prunusspecies分离的裂解酶相反，它不是一个黄素蛋白。在十二烷基硫酸钠存在下通过凝胶电泳估计亚基分子量为42，000。通过凝胶过滤（HPLC）估计酶的天然分子量为82，000。该酶具有约5.5的窄pH最适值，并且在4 °C下高度稳定。
The hydroxynitrile lyase (EC 4.1.2._) which catalyzes the dissociation of the cyanohydrins of acetone and 2-butanone has been isolated and purified from young seedlings of flax (Linum usitatissimumL.). The purification procedure involved precipitation with (NH4)2SO4, chromatofocusing, and chromatography on DEAE-cellulose, hydroxylapatite, Sephacryl 200, and Matrex Red A gel columns with a final recovery of 21%. Purification of 136-fold yielded an apparently homogeneous preparation that, in contrast to the lyases isolated fromPrunusspecies, is not a flavoprotein. The subunit molecular weight of 42,000 was estimated by gel electrophoresis in the presence of sodium dodecyl sulfate. The native molecular weight of the enzyme was estimated by gel filtration (HPLC) to be 82,000. The enzyme has a narrow pH optimum around 5.5 and is highly Stable at 4 °C.