Molecular Basis for Mycophenolic Acid Biosynthesis in Penicillium brevicompactum
Molecular Basis for Mycophenolic Acid Biosynthesis in Penicillium brevicompactum
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DOI：
10.1128/aem.03015-10
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发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Nielsen, Jens
中科院分区：
文献类型：
--
作者：
Regueira, Torsten Bak;Kildegaard, Kanchana Rueksomtawin;Nielsen, Jens
霉酚酸（MPA）是越来越重要的免疫抑制药物CellCept（罗氏）和Myfortic（诺华）的活性成分。尽管MPA的历史悠久，但其生物合成的分子基础仍然是个谜。在这里，我们报告的聚酮合酶（PKS），MpaC，我们成功地表征和确定为负责MPA生产短密青霉菌的发现。mpaC最可能存在于短密青霉基因组中的25-kb基因簇中。通过靶向假定的抗性基因，在这种情况下，编码IMP脱氢酶（IMPDH）的基因的额外拷贝，成功地定位了基因簇。通过缺失短密青霉聚酮合成酶基因mpaC，克隆了短密青霉MPA生物合成基因簇，并对其进行了生物信息学分析。正如预期的那样，基因缺失完全消除了MPA的产生以及来自短密青霉MPA生物合成途径的几种其他代谢产物的产生。我们的工作为通过代谢工程生产MPA和类似物奠定了基础。
Mycophenolic acid (MPA) is the active ingredient in the increasingly important immunosuppressive pharmaceuticals CellCept (Roche) and Myfortic (Novartis). Despite the long history of MPA, the molecular basis for its biosynthesis has remained enigmatic. Here we report the discovery of a polyketide synthase (PKS), MpaC, which we successfully characterized and identified as responsible for MPA production in Penicillium brevicompactum. mpaC resides in what most likely is a 25-kb gene cluster in the genome of Penicillium brevicompactum. The gene cluster was successfully localized by targeting putative resistance genes, in this case an additional copy of the gene encoding IMP dehydrogenase (IMPDH). We report the cloning, sequencing, and the functional characterization of the MPA biosynthesis gene cluster by deletion of the polyketide synthase gene mpaC of P. brevicompactum and bioinformatic analyses. As expected, the gene deletion completely abolished MPA production as well as production of several other metabolites derived from the MPA biosynthesis pathway of P. brevicompactum. Our work sets the stage for engineering the production of MPA and analogues through metabolic engineering.