Joblin K N, Johnson A W, Lappert M F, Wallis O C
Biochim Biophys Acta. 1976 Nov 8;452(1):262-70. doi: 10.1016/0005-2744(76)90079-6.
Purification of ethanolamine ammonia-lyase (EC 4.3.1.7) from a Clostridium sp. grown at the University of Sussex, U.K. and the National Institutes of Health, U.S.A., has been compared and an improved isotopic assay for the enzyme has been developed. Successful purification of this enzyme from Sussex-grown cells requires modification of the published procedure (Kaplan and Stadtman (1968) J. Biol, Chem. 243, 1787-1793) principally a 70% decrease in volume during precipitation with 0.4 M NaCl. This modification also increases the yield from N.I.H.-grown cells. Purified enzyme, resolved of inactive cobalamins, has the same high specific activity from both sources and behaves in the same way on disc gel electrophoresis. Sussex enzyme, before resolution, has less than 20% of the specific activity of unresolved N.I.H. enzyme and contains over 50% more inactive cobalamin. The bound cobalamin from both preparations has been identified as a "base-on" Co11 psi-cobalamin.
从一株在英国苏塞克斯大学和美国国立卫生研究院培养的梭菌属细菌中纯化乙醇胺氨裂解酶（EC 4.3.1.7），并对其进行了比较，同时开发了一种改进的该酶同位素检测方法。要从在苏塞克斯培养的细胞中成功纯化这种酶，需要对已发表的方法（卡普兰和斯塔特曼（1968年）《生物化学杂志》243卷，1787 - 1793页）进行修改，主要是在用0.4 M氯化钠沉淀过程中体积减少70%。这种修改也提高了从美国国立卫生研究院培养的细胞中的产量。去除无活性钴胺素的纯化酶，来自这两种来源的都具有相同的高比活性，并且在圆盘凝胶电泳中的行为相同。在分离之前，苏塞克斯的酶的比活性不到未分离的美国国立卫生研究院的酶的20%，并且含有比无活性钴胺素多50%以上。两种制剂中结合的钴胺素已被鉴定为“碱基朝上”的Co11ψ - 钴胺素。
