Biosynthesis of methyl-proline containing griselimycins, natural products with anti-tuberculosis activity.
Biosynthesis of methyl-proline containing griselimycins, natural products with anti-tuberculosis activity.
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DOI：
10.1039/c7sc02622f
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发表时间：
2017-11-01
期刊：
影响因子：
8.4
通讯作者：
Müller R
中科院分区：
文献类型：
--
作者：
Lukat P;Katsuyama Y;Wenzel S;Binz T;König C;Blankenfeldt W;Brönstrup M;Müller R
通过生物化学和结构分析，报道了链霉菌DSM 40835中雷司利霉素的生物合成和L亮氨酸立体定向转化为(2S，4R)-4-甲基-脯氨酸的途径。格里利霉素(Grislimycins，GM)是一类具有极强抗结核活性的多肽。它们含有多达三种(2S，4R)-4-甲基-脯氨酸(4-MePro)，其中一种可阻止氧化降解，并在动物模型中增加代谢稳定性。具有这种取代作用的天然同系物在发酵培养中只是一个很小的成分。我们发现在反应中加入4-MePro可以显著提高该产物的产量，并对4-MePro生物合成和掺入的分子基础进行了研究。我们鉴定了GM生物合成基因簇编码一个非核糖体多肽合成酶和一个4-MePro形成的亚操纵子。利用异源表达、基因失活和体外实验，我们证明了4-MePro是通过亮氨酸羟基化、氧化成醛、环合以及随后的还原生成的。亮氨酸羟基酶GRIE与底物和产物的络合物的晶体结构已经确定，这为该反应的立体专一性提供了洞察。
The biosynthesis of griselimycins in Streptomyces DSM 40835 and the pathway that stereospecifically converts l-leucine to (2S,4R)-4-methyl-proline are reported by means of biochemical and structural analysis. Griselimycins (GMs) are depsidecapeptides with superb anti-tuberculosis activity. They contain up to three (2S,4R)-4-methyl-prolines (4-MePro), of which one blocks oxidative degradation and increases metabolic stability in animal models. The natural congener with this substitution is only a minor component in fermentation cultures. We showed that this product can be significantly increased by feeding the reaction with 4-MePro and we investigated the molecular basis of 4-MePro biosynthesis and incorporation. We identified the GM biosynthetic gene cluster as encoding a nonribosomal peptide synthetase and a sub-operon for 4-MePro formation. Using heterologous expression, gene inactivation, and in vitro experiments, we showed that 4-MePro is generated by leucine hydroxylation, oxidation to an aldehyde, and ring closure with subsequent reduction. The crystal structures of the leucine hydroxylase GriE have been determined in complex with substrates and products, providing insight into the stereospecificity of the reaction.
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影响因子：
15
作者：
Riggs-Gelasco, PJ;Price, JC;Krebs, C
通讯作者：
Krebs, C
影响因子：
64.8
作者：
Valegård, K;van Scheltinga, ACT;Andersson, I
通讯作者：
Andersson, I
影响因子：
14.8
作者：
Kelley, Lawrence A.;Sternberg, Michael J. E.
通讯作者：
Sternberg, Michael J. E.
影响因子：
14.9
作者：
DEUTCH, AH;SMITH, CJ;KRETSCHMER, PJ
通讯作者：
KRETSCHMER, PJ
影响因子：
3.6
作者：
Luesch, H;Hoffmann, D;Moore, RE
通讯作者：
Moore, RE