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地衣黄粉衣中聚酮合酶基因的克隆及异源转录

Cloning and heterologous transcription of a polyketide synthase gene from the lichen Solorina crocea.

作者信息

Gagunashvili Andrey N, Davídsson Snorri P, Jónsson Zophonías O, Andrésson Olafur S

机构信息

Institute of Biology, University of Iceland, Sturlugata 7, IS-101 Reykjavik, Iceland.

出版信息

Mycol Res. 2009 Mar;113(Pt 3):354-63. doi: 10.1016/j.mycres.2008.11.011. Epub 2008 Dec 6.

Abstract

Lichens and most ascomycete fungi produce polyketide secondary metabolites often with valuable biological activities. Their biosynthesis is primarily governed by large iterative multifunctional type I polyketide synthases. Although there has been good progress studying filamentous non-lichenized fungi, there is limited information on polyketide biosynthesis in lichens and their mycobionts, due to their slow growth, difficulties in establishing pure cultures, and the absence of methods for direct genetic manipulation. However, heterologous expression in a surrogate host offers an alternative approach for exploring lichen polyketide biosynthesis. Here, we report cloning of a type I polyketide synthase gene from the foliose lichen Solorina crocea and its heterologous transcription in the filamentous fungus Aspergillus oryzae, including processing of the transcript. No new polyketide product was detected. The lichen polyketide synthase showed greatest homology with uncharacterized genes from filamentous fungi and lower homology with proteins catalysing biosynthesis of the decaketide alternapyrone and the tetraketide side-chain of squalestatin. The technology platform utilized here presents a useful tool for functional characterization of fungal biosynthetic genes and provides a means for novel production of valuable compounds.

摘要

地衣和大多数子囊菌会产生通常具有重要生物活性的聚酮类次生代谢产物。它们的生物合成主要由大型迭代多功能I型聚酮合酶控制。尽管在研究丝状非地衣化真菌方面取得了良好进展,但由于地衣及其共生菌生长缓慢、难以建立纯培养物以及缺乏直接基因操作方法,关于地衣及其共生菌中聚酮生物合成的信息有限。然而,在替代宿主中的异源表达为探索地衣聚酮生物合成提供了一种替代方法。在此,我们报告了从叶状地衣环裂石耳中克隆I型聚酮合酶基因及其在丝状真菌米曲霉中的异源转录,包括转录本的加工。未检测到新的聚酮产物。地衣聚酮合酶与丝状真菌中未表征的基因具有最高的同源性,与催化十酮交替吡喃酮和角鲨烯他汀四酮侧链生物合成的蛋白质同源性较低。这里使用的技术平台为真菌生物合成基因的功能表征提供了有用的工具,并为有价值化合物的新生产提供了一种手段。

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