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免疫抑制剂 Brasilicardin:在异源宿主 Amycolatopsis japonicum 中确定生物合成基因簇。

The Immunosuppressant Brasilicardin: Determination of the Biosynthetic Gene Cluster in the Heterologous Host Amycolatopsis japonicum.

机构信息

Microbiology/Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine (IMIT), Eberhard Karls University of Tübingen, Tübingen, Germany.

Department of Pharmaceutical Biology, Pharmaceutical Institute, Eberhard Karls University of Tübingen, Tübingen, Germany.

出版信息

Biotechnol J. 2018 Feb;13(2). doi: 10.1002/biot.201700527. Epub 2017 Nov 13.

Abstract

Nocardia terpenica IFM 0406 is the producer of the immunosuppressants brasilicardins A-D. Brasilicardin is a promising compound because of its unique mode of action and its higher potency and reduced toxicity compared to today's standard drugs. However, production of brasilicardin is so far hampered as Nocardia terpenica IFM 0406 synthesizes brasilicardin in only low amounts and represents a human pathogen (biosafety level 2 BSL2). In order to achieve a safe and high yield production of brasilicardin A (BraA), the authors heterologously express the brasilicardin gene cluster in the nocardioform actinomycete Amycolatopsis japonicum (A. japonicum::bcaAB01), which is fast growing, genetically accessible and closely related to N. terpenica IFM 0406. In A. japonicum::bcaAB01, four brasilicardin congeners, intermediates of the BraA biosynthesis, are produced. Investigation of the genes flanking the previously defined brasilicardin biosynthetic gene cluster revealed two novel genes (bra0, bra12), which are involved in brasilicardin biosynthesis: bra12 encodes a transcriptional activator of the brasilicardin gene cluster. bra0 codes for a dioxygenase involved in methoxylation of brasilicardin. Based on this finding the authors are able to revise the proposed brasilicardin biosynthesis.

摘要

松萝诺卡氏菌 IFM 0406 是免疫抑制剂巴西卡丁 A-D 的产生菌。巴西卡丁是一种很有前途的化合物,因为它具有独特的作用模式,而且与目前的标准药物相比,其效力更高,毒性更低。然而,由于松萝诺卡氏菌 IFM 0406 的产量很低,而且它是一种人类病原体(生物安全级别 2 BSL2),因此巴西卡丁的生产迄今为止一直受到阻碍。为了安全高效地生产巴西卡丁 A(BraA),作者在诺卡氏放线菌 Amycolatopsis japonicum(A. japonicum::bcaAB01)中异源表达巴西卡丁基因簇,A. japonicum 生长迅速,遗传可及性高,与松萝诺卡氏菌 IFM 0406 密切相关。在 A. japonicum::bcaAB01 中,产生了四个巴西卡丁同系物,它们是 BraA 生物合成的中间体。对先前定义的巴西卡丁生物合成基因簇侧翼基因的研究揭示了两个新基因(bra0、bra12),它们参与巴西卡丁生物合成:bra12 编码巴西卡丁基因簇的转录激活子。bra0 编码参与巴西卡丁甲氧基化的双加氧酶。基于这一发现,作者能够修正巴西卡丁生物合成的提议。

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