林可霉素 A 生物合成中八磷酸中间物的构建：LmbR 和 LmbN 催化反应的表征。
Construction of the octose 8-phosphate intermediate in lincomycin A biosynthesis: characterization of the reactions catalyzed by LmbR and LmbN.
机构信息
Division of Medicinal Chemistry, College of Pharmacy, and Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas 78712, USA.
出版信息
J Am Chem Soc. 2012 Oct 24;134(42):17432-5. doi: 10.1021/ja308221z. Epub 2012 Oct 11.
Abstract
Lincomycin A is a potent antimicrobial agent noted for its unusual C1 methylmercapto-substituted 8-carbon sugar. Despite its long clinical history for the treatment of Gram-positive infections, the biosynthesis of the C(8)-sugar, methylthiolincosamide (MTL), is poorly understood. Here, we report our studies of the two initial enzymatic steps in the MTL biosynthetic pathway leading to the identification of D-erythro-D-gluco-octose 8-phosphate as a key intermediate. Our experiments demonstrate that this intermediate is formed via a transaldol reaction catalyzed by LmbR using D-fructose 6-phosphate or D-sedoheptulose 7-phosphate as the C(3) donor and D-ribose 5-phosphate as the C(5) acceptor. Subsequent 1,2-isomerization catalyzed by LmbN converts the resulting 2-keto C(8)-sugar (octulose 8-phosphate) to octose 8-phosphate. These results provide, for the first time, in vitro evidence for the biosynthetic origin of the C(8) backbone of MTL.
摘要
林可霉素 A 是一种强效的抗菌药物，其特点是 C1 位上有一个不寻常的甲基巯基取代的 8 碳糖。尽管它在治疗革兰氏阳性感染方面有着悠久的临床历史，但对于 C(8)-糖，即甲基硫代林肯酰胺（MTL）的生物合成过程仍知之甚少。在这里，我们报告了我们对 MTL 生物合成途径中最初的两个酶促步骤的研究，这些研究确定了 D-erythro-D-gluco-octose 8-phosphate 作为关键中间体。我们的实验表明，该中间体是通过 LmbR 催化的转醛醇反应形成的，该反应使用 D-果糖 6-磷酸或 D-sedoheptulose 7-磷酸作为 C(3)供体，D-核糖 5-磷酸作为 C(5)受体。随后由 LmbN 催化的 1,2-异构化将生成的 2-酮 C(8)-糖（辛糖 8-磷酸）转化为辛糖 8-磷酸。这些结果首次提供了 MTL 的 C(8)骨架生物合成来源的体外证据。
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1
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Chembiochem. 2011 Jul 4;12(10):1454-74. doi: 10.1002/cbic.201100072. Epub 2011 May 13.
2
Fermenting next generation glycosylated therapeutics.发酵下一代糖基化治疗药物。
ACS Chem Biol. 2011 Jan 21;6(1):14-7. doi: 10.1021/cb100375y.
3
Mechanistic studies of the biosynthesis of 2-thiosugar: evidence for the formation of an enzyme-bound 2-ketohexose intermediate in BexX-catalyzed reaction.2-硫代糖生物合成的机制研究：BexX 催化反应中酶结合 2-酮己糖中间产物形成的证据。
J Am Chem Soc. 2010 Nov 10;132(44):15544-6. doi: 10.1021/ja108061c.
4
Kdo: a critical monosaccharide for bacteria viability.Kdo：细菌生存能力的关键单糖。
Nat Prod Rep. 2010 Nov;27(11):1618-29. doi: 10.1039/c004750n. Epub 2010 Sep 29.
5
A biosynthetic pathway for BE-7585A, a 2-thiosugar-containing angucycline-type natural product.一种含有 2-硫代糖的安卡环类天然产物 BE-7585A 的生物合成途径。
J Am Chem Soc. 2010 Jun 2;132(21):7405-17. doi: 10.1021/ja1014037.
6
Natural products version 2.0: connecting genes to molecules.天然产物 2.0 版：将基因与分子联系起来。
J Am Chem Soc. 2010 Mar 3;132(8):2469-93. doi: 10.1021/ja909118a.
7
Transaldolase: from biochemistry to human disease.转醛醇酶：从生物化学到人类疾病
Int J Biochem Cell Biol. 2009 Jul;41(7):1482-94. doi: 10.1016/j.biocel.2009.02.001. Epub 2009 Feb 11.
8
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