Heterologous Expression of the Biosynthetic Gene Clusters of Coumermycin A1, Clorobiocin and Caprazamycins in Genetically Modified Streptomyces coelicolor Strains
Heterologous Expression of the Biosynthetic Gene Clusters of Coumermycin A1, Clorobiocin and Caprazamycins in Genetically Modified Streptomyces coelicolor Strains
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DOI：
10.1002/bip.21493
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发表时间：
2010-09-01
期刊：
影响因子：
2.9
通讯作者：
Heide, Lutz
中科院分区：
文献类型：
--
作者：
Flinspach, Katrin;Westrich, Lucia;Heide, Lutz
氨基香豆素抗生素氯生菌素和香豆霉素 A(1) 以及脂核苷抗生素卡普拉霉素的生物合成基因簇稳定整合到源自天蓝色链霉菌 A3(2) 的不同宿主菌株的基因组中。对于氯霉素衍生物在化学成分确定的培养基中的异源表达，在生长培养基中添加 0.6% 的甲硅烷氧基化环氧乙烷/环氧丙烷共聚物 Q2-5247 被证明可导致生产率提高 4.8 倍。据推测，该共聚物充当氧载体。额外加入氯化钴 (0.2-2 mg l(-1)) 显着增加了所需化合物氯霉素在总生产的氯霉素衍生物中的百分比。这很可能是由于氯霉素生物合成酶 CloN6 催化的钴胺素依赖性甲基化反应受到刺激。所有三个研究的宿主菌株（S. coelicolor M512、M1146 和 M1154）均产生相似的总氯霉素衍生物生产率（在 0.6% Q2-5247 和 0.2 mg l(-1) CoCl2 存在下平均为 158 mg l(-1) I）。相比之下，卡普拉霉素衍生物的异源生产在菌株 M1154 中是最佳的（平均产量为 152 mg l(-1)）。 (C) 2010 年 Wiley 期刊公司。生物聚合物 93：823-832，2010。
The biosynthetic gene clusters of the aminocoumarin antibiotics clorobiocin and coumermycin A(1) and of the liponucleoside antibiotic caprazamycin were stably integrated into the genomes of different host strains derived from Streptomyces coelicolor A3(2). For the heterologous expression of clorobiocin derivatives in a chemically defined medium, inclusion of 0.6% of the siloxylated ethylene oxide/propylene oxide copolymer Q2-5247 into the growth medium proved to result in a 4.8-fold increase of productivity. Presumably, this copolymer acts as an oxygen carrier. The additional inclusion of cobalt chloride (0.2-2 mg l(-1)) dramatically increased the percentage of the desired compound clorobiocin within the total produced clorobiocin derivatives. This is very likely due to a stimulation of a cobalamin-dependent methylation reaction catalyzed by the enzyme CloN6 of clorobiocin biosynthesis. All three investigated host strains (S. coelicolor M512, M1146 and M1154) gave similar production rates of total clorobiocin derivatives (on average, 158 mg l(-1) I in the presence of 0.6% Q2-5247 and 0.2 mg l(-1) CoCl2). In contrast, heterologous production of caprazamycin derivatives was optimal in strain M1154 (amounts of 152 mg l(-1) on average). (C) 2010 Wiley Periodicals, Inc. Biopolymers 93: 823-832, 2010.