Precursor-directed biosynthesis of 6-deoxyerythronolide B analogs in Streptomyces coelicolor:: Understanding precursor effects
Precursor-directed biosynthesis of 6-deoxyerythronolide B analogs in Streptomyces coelicolor:: Understanding precursor effects
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DOI：
10.1021/bp000063l
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发表时间：
2000-07-01
影响因子：
2.9
通讯作者：
Licari, P
中科院分区：
文献类型：
--
作者：
Leaf, T;Cadapan, L;Licari, P
正在开发一种采用底物定向生物合成的发酵方法，用于生产6-脱氧阿糖胞苷B（6-dEB）类似物。通过天蓝色链霉菌中基于质粒的系统，6-dEB合成由6-dEB合酶（DEBS）催化。6-通过DEBS的酮合酶（KS）1结构域的失活来消除dEB合成，但是可以通过提供合成的活化的二酮化合物来恢复dEB合成。由于其固有的催化灵活性，KS 1缺陷型DEBS能够利用非天然的二酮化合物形成各种13-取代的6-dEB。在这里，我们表征与双酮化合物进料摇瓶实验中的工艺变量。发现13-R-6-dEB的产生强烈依赖于二酮化合物进料浓度、进料时培养物的生长阶段以及二酮化合物部分中存在的R-基团。在所有情况下，大部分的进料双酮化合物被细胞降解。
A fermentation process employing precursor-directed biosynthesis is being developed for the manufacture of 6-deoxyerythronolide B (6-dEB) analogues. Through a plasmid-based system in Streptomyces coelicolor, 6-dEB synthesis is catalyzed by 6-dEB synthase (DEBS). 6-dEB synthesis is abolished by inactivation of the ketosynthase (KS) 1 domain of DEBS but can be restored by providing synthetic activated diketides. Because of its inherent catalytic flexibility, the KS1-deficient DEBS is capable of utilizing unnatural diketides to form various 13-substituted 6-dEBs. Here we characterize process variables associated with diketide feeding in shake-flask experiments. 13-R-6-dEB production was found to depend strongly on diketide feed concentrations, on the growth phase of cultures at feeding time, and on the R-group present in the diketide moiety. In all cases a major portion of the fed diketides was degraded by the cells.