聚乙烯生物合成中的聚乙烯链延长和甲基组表皮化机制和立体化学
Xinqiang Xie1, Ashish Garg1, Chaitan Khosla2
1Department of Chemistry, Brown University , Box H, Providence, Rhode Island 02912-9108, United States.
在聚乙烯抗生素生物合成中,氧降解酶 (KR0) 域充当表皮酶. 这些域催化了 (2R) -2-甲基-3-基-ACP中间体的转化,挑战了关于它们功能的先前假设.
科学领域:
- 生物化学
- 分子生物学
- 有机化学
背景情况:
- 像南菌素和沙利诺菌素这样的多抗生素是由称为多基合成酶 (PKS) 的大型酶复合体合成的.
- 在这些PKS中,某些被指定为KR0的基因减少酶 (KR) 域已知是氧化还原不活跃的.
- 这些KR0域被假设在抗生素生物合成过程中起作用.
研究的目的:
- 来自南菌素和盐酸菌素合成酶的氧化还原酶 (KR0) 域的基质特异性和表皮酶活性.
- 提供KR0域在产生特定中间体中的提议表皮酶功能的直接证据.
主要方法:
- 开发结合合成酶 (KS) - 减少酶 (KR) 试验法,以分析特定PKS领域的产物.
- 使用标记基质进行平衡同位素交换实验.
- 在酶试验中使用了来自南菌素和盐酸菌素合成酶的重组氧化-无活性KR0域.
主要成果:
- 结合KS-KR试验证实,Nan[KS1][AT1]二域仅产生 (2R) -2-甲基-3- pentanoyl-ACP中间体.
- 同位素交换实验表明,KR0域 (NanKR10,NanKR50,SalKR70) 催化了标记的基-基-ACP前体的洗.
- 这种洗归因于KR0催化过渡形成的酸-ACP氧化产物的表皮化.
结论:
- 聚乙烯合成酶的氧化无活性KR0域作为表皮酶起作用,而不仅仅是结构性或无活性成分.
- 这些KR0域特异性地表化 (2R) -2-甲基-3-基-ACP中间体,这是自然基质.
- KR0域的表皮酶活性独立于通常参与缩酶催化表皮酶的保存Tyr-Ser二.
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