公开(公告)号：US4770781A

公开(公告)日：1988-09-13

申请号：US80182

申请日：1987-07-29

申请人： Jack A. Schmidt ,  Patricia M. Cameron ,  Guadalupe A. Limjuco

发明人： Jack A. Schmidt ,  Patricia M. Cameron ,  Guadalupe A. Limjuco

IPC分类号： B01D15/08 ,  B01J41/20 ,  C07K14/545

CPC分类号： B01J41/20 ,  B01D15/363 ,  C07K14/545 ,  B01D15/325 ,  B01D15/34

摘要： Human interleukin-1 (IL-1) charged species (pI 6.8, 6.0, 5.4 and 5.2) are separated and purified by a rapid and efficient two or three stage chromatographic process. Interleukin-1 species present in concentrated culture fluid from stimulated human mononuclear cells are separated and partially purified by anion exchange high performance liquid chromatography. The pI 5.4 and 5.2 species in addition are partially purified by HPLC gel filtration chromatography. Each of the individual IL-1 species are then finally purified by reverse phase high performance liquid chromatography. This process results in homogenously pure pI 6.8, pI 5.4 and pI 5.2 species while the pI 6.0 species is isolated in a substantially pure form.

摘要翻译： 通过快速和有效的两级或三级色谱法分离和纯化人白细胞介素-1（IL-1）带电物种（pI 6.8,6.0,5.4和5.2）。 通过阴离子交换高效液相色谱分离来自刺激的人单核细胞的浓缩培养液中存在的白细胞介素-1物质。 另外通过HPLC凝胶过滤色谱法部分纯化pI 5.4和5.2物质。 然后通过反相高效液相色谱将每种单独的IL-1物质进行纯化。 该过程产生均匀纯的pI 6.8，pI 5.4和pI 5.2物种，而pI 6.0物质以基本上纯的形式分离。

公开(公告)号：US5264355A

公开(公告)日：1993-11-23

申请号：US908244

申请日：1992-07-02

申请人： Ali Shafiee ,  Patricia M. Cameron ,  David A. Boulton ,  Louis Kaplan ,  Haideh Motamedi

发明人： Ali Shafiee ,  Patricia M. Cameron ,  David A. Boulton ,  Louis Kaplan ,  Haideh Motamedi

IPC分类号： C12N9/10 ,  C12N9/08

CPC分类号： C12N9/1007

摘要： Described is a new enzyme, 31-O-desmethyl-FK-506 O-methyltransferase (FKMT) and its N-terminal amino acid sequence. This enzyme can specifically and preferentially methylate the C-31 hydroxy group in 31-O-desmethyl FK-506/FR-900506 and other 31-O-desmethyl analogs of FK-506/FR-900506 and related compounds such as 31-O-desmethyl immunomycin type molecules, also designated FK-520 and FR-900520 by Fujisawa. The enzyme can be isolated from a FK-506 producing microorganism Streptomyces sp. (Merck Culture Collection No. MA 6858) ATCC No. 55098. Employing the enzyme in its active form, supplemented with Mg.sup.+2 ion, in the presence of the methyl donor, S-adenosyl methionine (SAM), derivatives of 31-O-desmethyl FK-506, 31-O-desmethyl FK-520 and related 31-O-desmethyl compounds may be prepared.

摘要翻译： 描述了一种新的酶，31-O-去甲基-FK-506 O-甲基转移酶（FKMT）及其N-末端氨基酸序列。 该酶可以特异地和优先地甲基化31-O-脱甲基FK-506 / FR-900506中的C-31羟基和FK-506 / FR-900506的其它31-O-脱甲基类似物和相关化合物如31-O 也称为Fujisawa的FK-520和FR-900520。 该酶可以从FK-506产生的微生物链霉菌（Streptomyces sp。）中分离出来。 （Merck Culture Collection No.MA6858）ATCC No.55098.在甲基供体存在下使用活性形式的补充了Mg + 2离子的酶，S-腺苷甲硫氨酸（SAM），31-O的衍生物 - 甲基FK-506,31-O-去甲基FK-520和相关的31-O-脱甲基化合物。