Methods for the preparation of chlorophyllide a: An intermediate of the chlorophyll biosynthetic pathway
Methods for the preparation of chlorophyllide a: An intermediate of the chlorophyll biosynthetic pathway
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DOI：
10.1016/j.ab.2011.08.028
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发表时间：
2011-12-15
影响因子：
2.9
通讯作者：
Hansson, Mats
中科院分区：
文献类型：
--
作者：
Muller, Andre H.;Gough, Simon P.;Hansson, Mats
叶绿素a是叶绿素和细菌叶绿素生物合成后期的代谢产物。从Rhodobacter capsulatus CB 1200和大麦（Hordeum vulgare L.）描述和比较。R. capsulatus CB 1200是细菌叶绿素a生物合成途径中的双突变体，并且当在含吐温80的液体培养基中生长时，叶绿素a由细胞分泌。通过液相或固相萃取纯化，从1L培养物中得到7mg叶绿酸a。在第二种方法中，内在叶绿素酶活性被用来在野生型或叶绿素b缺陷大麦的叶的丙酮制剂中使叶绿素脱植酸。通过液相萃取实现纯化，每克大麦叶产生14微克叶绿酸a。通过薄层色谱法、吸收光谱法和质谱法鉴定叶绿素a。(C)2011 Elsevier Inc. All rights reserved.
Chlorophyllide a is a metabolite late in the biosynthesis of chlorophylls and bacteriochlorophylls. Isolation procedures for chlorophyllide a from Rhodobacter capsulatus CB1200 and barley (Hordeum vulgare L.) are described and compared. R. capsulatus CB1200 is a double mutant in the bacteriochlorophyllide a biosynthetic pathway, and chlorophyllide a is excreted by the cells when grown in Tween 80-containing liquid medium. It was purified by liquid or solid phase extraction, yielding 7 mg of chlorophyllide a from 1 L of culture. In a second approach, intrinsic chlorophyllase activity was used to dephytylate chlorophyll in an acetonic preparation of leaves of wild-type or chlorophyll b-deficient barley. Purification was achieved by liquid phase extraction, yielding 14 mu g of chlorophyllide a per gram of barley leaves. Chlorophyllide a was identified by thin layer chromatography, absorption spectroscopy, and mass spectrometry. (C) 2011 Elsevier Inc. All rights reserved.