Establishing an Efficient Genetic Manipulation System for Sulfated Echinocandin Producing Fungus Coleophoma empetri.
Establishing an Efficient Genetic Manipulation System for Sulfated Echinocandin Producing Fungus Coleophoma empetri.
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DOI：
10.3389/fmicb.2021.734780
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发表时间：
2021
影响因子：
5.2
通讯作者：
Lu X
中科院分区：
文献类型：
--
作者：
Men P;Wang M;Li J;Geng C;Huang X;Lu X
米卡芬净是治疗侵袭性真菌感染的重要棘白菌素类抗真菌药。在工业上，米卡芬净来源于天然产物FR 901379，FR 901379是由丝状真菌Coleophoma empetri产生的非核糖体环状六肽。C.遗传操作的困难。empetri限制了FR 901379生物合成机制的阐明。在这项工作中，我们开发了一个有效的遗传操作系统中的工业FR 901379生产菌株C。empetri MEFC 009.首先，建立了一种简便的原生质体介导的转化方法。其次，通过这种转化方法，验证了必需的遗传因子。选择标记hph、neo和nat可用于转化，启动子Ppgk、PgpdA和PgpdAt在C. empetri MEFC 009.第三，通过删除ku 80基因，同源重组的频率从4%提高到100%，从而产生用于基因打靶的优良底盘细胞。此外，该遗传操作系统的优势被证明在鉴定的聚酮合酶（PKS）负责二羟基萘（DHN）-黑色素的生物合成。该遗传操作系统将为FR 901379的研究和进一步的C.恩佩特里
Micafungin is an important echinocandin antifungal agent for the treatment of invasive fungal infections. In industry, micafungin is derived from the natural product FR901379, which is a non-ribosomal cyclic hexapeptide produced by the filamentous fungus Coleophoma empetri. The difficulty of genetic manipulation in C. empetri restricts the clarification of FR901379 biosynthetic mechanism. In this work, we developed an efficient genetic manipulation system in the industrial FR901379-producing strain C. empetri MEFC009. Firstly, a convenient protoplast-mediated transformation (PMT) method was developed. Secondly, with this transformation method, the essential genetic elements were verified. Selectable markers hph, neo, and nat can be used for the transformation, and promotors Ppgk, PgpdA, and PgpdAt are functional in C. empetri MEFC009. Thirdly, the frequency of homologous recombination was improved from 4 to 100% by deleting the ku80 gene, resulting in an excellent chassis cell for gene-targeting. Additionally, the advantage of this genetic manipulation system was demonstrated in the identification of the polyketide synthase (PKS) responsible for the biosynthesis of dihydroxynapthalene (DHN)-melanin. This genetic manipulation system will be a useful platform for the research of FR901379 and further genome mining of secondary metabolites in C. empetri.
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影响因子：
3.3
作者：
Marcone, Giorgia Letizia;Carrano, Lucia;Beltrametti, Fabrizio
通讯作者：
Beltrametti, Fabrizio
影响因子：
--
作者：
Roth, Mitchell G.;Chilvers, Martin I.
通讯作者：
Chilvers, Martin I.
影响因子：
2.1
作者：
Moriwaki, A;Kihara, J;Honda, Y
通讯作者：
Honda, Y
嗜热丝状真菌嗜热毁丝霉的代谢工程产生富马酸
影响因子：
6.3
作者：
Gu, Shuying;Li, Jingen;Tian, Chaoguang
通讯作者：
Tian, Chaoguang
DOI：
10.1007/s10295-013-1385-0
发表时间：
2014-03-01
影响因子：
3.4
作者：
Huang, Xuenian;Lu, Xuefeng;Li, Jian-Jun
通讯作者：
Li, Jian-Jun