Engineered biosynthesis of pimaricin derivatives with improved antifungal activity and reduced cytotoxicity
Engineered biosynthesis of pimaricin derivatives with improved antifungal activity and reduced cytotoxicity
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具有改善抗真菌活性和降低细胞毒性的匹马星衍生物的工程生物合成
DOI：
10.1007/s00253-015-6635-9
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发表时间：
2015-08-01
影响因子：
5
通讯作者：
Bai, Linquan
中科院分区：
文献类型：
--
作者：
Qi, Zhen;Kang, Qianjin;Bai, Linquan
匹马菌素是一种重要的抗真菌抗生素，用于抗真菌治疗和预防食品工业中的霉菌污染。本研究通过Ginchattanoogensis L10链霉菌P450单加氧酶基因失活，合成了三个新的匹马菌素衍生物：12-脱羧-12-甲基匹马菌素（1）、4，5-脱环氧-12-脱羧-12-甲基匹马菌素（2）和2-氢-3-羟基-4，5-脱环氧-12-脱羧-12-甲基匹马菌素（3）。与匹马菌素相比，1对白色念珠菌ATCC 14053的抗真菌活性增加2倍，对红细胞的细胞毒性降低4.5倍，2具有相当的抗真菌活性和降低的细胞毒性，而3几乎没有抗真菌和溶血活性。遗传和生化分析证明，1是由2通过P450单加氧酶ScnD转化而来。因此，过量表达scnDinscnG-null菌株消除了2的积累，使1的产量提高了20%。而scnG/scnD双突变则使1的产量下降，2的产量提高了2.3倍。这些结果表明，通过基因工程获得的具有改进药理学性质的匹马菌素衍生物可以进一步开发为具有潜在临床应用价值的抗真菌药物。
Pimaricin is an important antifungal antibiotic for antifungal therapy and prevention of mould contamination in the food industry. In this study, three new pimaricin derivatives, 12-decarboxy-12-methyl pimaricin (1), 4,5-desepoxy-12-decarboxy-12-methyl pimaricin (2), and 2-hydro-3-hydroxy-4,5-desepoxy-12-decarboxy-12-methyl pimaricin (3), were generated through the inactivation of P450 monooxygenase genescnGinStreptomyces chattanoogensisL10. Compared with pimaricin,1displayed a twofold increase in antifungal activity againstCandida albicansATCC 14053 and a 4.5-fold decrease in cytotoxicity with erythrocytes, and2had comparable antifungal activity and reduced cytotoxicity, whereas3showed nearly no antifungal and hemolytic activities. Genetic and biochemical analyses proved that1is converted from2by P450 monooxygenase ScnD. Therefore, the overexpression ofscnDinscnG-null strain eliminated the accumulation of2and improved the yield of1by 20 %. Conversely,scnG/scnDdouble mutation abolished the production of1and improved the yield of2to 2.3-fold. These results indicate that the pimaricin derivatives with improved pharmacological properties obtained by genetic engineering can be further developed into antifungal agents for potential clinical application.