• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

建立用于生产硫酸化棘白菌素的真菌的高效基因操作体系

Establishing an Efficient Genetic Manipulation System for Sulfated Echinocandin Producing Fungus .

作者信息

Men Ping, Wang Min, Li Jinda, Geng Ce, Huang Xuenian, Lu Xuefeng

机构信息

Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.

Shandong Energy Institute, Qingdao, China.

出版信息

Front Microbiol. 2021 Aug 20;12:734780. doi: 10.3389/fmicb.2021.734780. eCollection 2021.

DOI:10.3389/fmicb.2021.734780
PMID:34489920
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8417879/
Abstract

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 . The difficulty of genetic manipulation in restricts the clarification of FR901379 biosynthetic mechanism. In this work, we developed an efficient genetic manipulation system in the industrial FR901379-producing strain MEFC009. Firstly, a convenient protoplast-mediated transformation (PMT) method was developed. Secondly, with this transformation method, the essential genetic elements were verified. Selectable markers , , and can be used for the transformation, and promotors , , and are functional in MEFC009. Thirdly, the frequency of homologous recombination was improved from 4 to 100% by deleting the 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 .

摘要

米卡芬净是一种用于治疗侵袭性真菌感染的重要棘白菌素类抗真菌药物。在工业上，米卡芬净来源于天然产物FR901379，它是由丝状真菌产生的一种非核糖体环状六肽。在[具体菌种]中进行基因操作的困难限制了对FR901379生物合成机制的阐明。在这项工作中，我们在工业生产FR901379的菌株MEFC009中开发了一种高效的基因操作系统。首先，开发了一种便捷的原生质体介导转化（PMT）方法。其次，利用这种转化方法，验证了关键的遗传元件。选择标记[具体标记1]、[具体标记2]和[具体标记3]可用于转化，启动子[具体启动子1]、[具体启动子2]和[具体启动子3]在MEFC009中具有功能。第三，通过缺失[具体基因]，同源重组频率从4%提高到了100%，从而产生了一个用于基因靶向的优良底盘细胞。此外，该基因操作系统在鉴定负责二羟基萘（DHN）-黑色素生物合成的聚酮合酶（PKS）方面的优势也得到了证明。这个基因操作系统将成为研究FR901379以及进一步挖掘[具体菌种]中次生代谢产物基因组的有用平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/2dda1406a550/fmicb-12-734780-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/310b2d276cb9/fmicb-12-734780-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/d4871f030843/fmicb-12-734780-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/c22bff3a777f/fmicb-12-734780-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/51d0b68ae5f3/fmicb-12-734780-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/4a83357f27ef/fmicb-12-734780-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/2dda1406a550/fmicb-12-734780-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/310b2d276cb9/fmicb-12-734780-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/d4871f030843/fmicb-12-734780-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/c22bff3a777f/fmicb-12-734780-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/51d0b68ae5f3/fmicb-12-734780-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/4a83357f27ef/fmicb-12-734780-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eab/8417879/2dda1406a550/fmicb-12-734780-g006.jpg

相似文献

1
Establishing an Efficient Genetic Manipulation System for Sulfated Echinocandin Producing Fungus .建立用于生产硫酸化棘白菌素的真菌的高效基因操作体系
Front Microbiol. 2021 Aug 20;12:734780. doi: 10.3389/fmicb.2021.734780. eCollection 2021.
2
Analysis of FR901379 Biosynthetic Genes in by Clustered Regularly Interspaced Short Palindromic Repeats/Cas9-Based Genomic Manipulation.通过基于簇状规则间隔短回文重复序列/ Cas9 的基因组操作分析 中的 FR901379 生物合成基因。
ACS Chem Biol. 2022 Aug 19;17(8):2130-2141. doi: 10.1021/acschembio.2c00250. Epub 2022 Jul 13.
3
Improving the production of the micafungin precursor FR901379 in an industrial production strain.提高工业生产菌株中棘白菌素前体 FR901379 的产量。
Microb Cell Fact. 2023 Mar 6;22(1):44. doi: 10.1186/s12934-023-02050-0.
4
Agrobacterium-tumefaciens-mediated transformation of antifungal-lipopeptide-producing fungus Coleophoma empetri F-11899.农杆菌介导的抗真菌脂肽产生菌Coleophoma empetri F-11899 的转化。
Curr Genet. 2009 Dec;55(6):623-30. doi: 10.1007/s00294-009-0275-5. Epub 2009 Oct 30.
5
Micafungin: a sulfated echinocandin.米卡芬净：一种硫酸化棘白菌素。
J Antibiot (Tokyo). 2009 Jan;62(1):27-35. doi: 10.1038/ja.2008.3. Epub 2009 Jan 9.
6
Biosynthesis mechanism, genome mining and artificial construction of echinocandin O-sulfonation.棘白素 O-硫酸化的生物合成机制、基因组挖掘和人工构建。
Metab Eng. 2022 Nov;74:160-167. doi: 10.1016/j.ymben.2022.10.006. Epub 2022 Nov 1.
7
Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907.通过 6907 号链霉菌生产 FR901379 酰化酶的菌株选育和放大发酵。
J Biosci Bioeng. 2011 Oct;112(4):409-14. doi: 10.1016/j.jbiosc.2011.06.002. Epub 2011 Jul 12.
8
Improvement of FR901379 production by mutant selection and medium optimization.通过突变体筛选和培养基优化提高FR901379的产量
J Biosci Bioeng. 2009 May;107(5):530-4. doi: 10.1016/j.jbiosc.2009.01.002.
9
Scale-up fermentation of echinocandin type antibiotic FR901379.大规模发酵棘白菌素类抗生素 FR901379。
J Biosci Bioeng. 2010 Feb;109(2):138-44. doi: 10.1016/j.jbiosc.2009.07.019. Epub 2009 Aug 27.
10
[Drug development from natural fermentation products: establishing a manufacturing process which maximizes the potential of microorganisms].[源于天然发酵产物的药物研发：建立最大化微生物潜能的制造工艺]
Yakugaku Zasshi. 2010 Nov;130(11):1471-8. doi: 10.1248/yakushi.130.1471.

引用本文的文献

1
Improving the production of micafungin precursor FR901379 in using heavy-ion irradiation and its mechanism analysis.利用重离子辐照提高棘白菌素前体FR901379的产量及其机制分析。
Mycology. 2024 Dec 12;16(2):941-955. doi: 10.1080/21501203.2024.2426484. eCollection 2025.
2
High-quality genome assembly and genetic transformation system of strain LTTK16-3, a fungal pathogen of Chinese hickory.中国山核桃真菌病原菌 LTTK16-3 的高质量基因组组装和遗传转化系统。
Microbiol Spectr. 2024 Mar 5;12(3):e0331123. doi: 10.1128/spectrum.03311-23. Epub 2024 Feb 13.
3
Improving the production of the micafungin precursor FR901379 in an industrial production strain.

本文引用的文献

1
CRISPR/Cas9-Based Genome Editing in the Filamentous Fungus and Its Application in Manipulating .基于 CRISPR/Cas9 的丝状真菌基因组编辑及其在调控中的应用 。
ACS Synth Biol. 2020 Aug 21;9(8):1968-1977. doi: 10.1021/acssynbio.9b00491. Epub 2020 Aug 12.
2
A protoplast generation and transformation method for soybean sudden death syndrome causal agents and .一种针对大豆猝死综合症致病因子的原生质体生成与转化方法以及……（原文此处不完整）
Fungal Biol Biotechnol. 2019 May 15;6:7. doi: 10.1186/s40694-019-0070-0. eCollection 2019.
3
antiSMASH 5.0: updates to the secondary metabolite genome mining pipeline.
提高工业生产菌株中棘白菌素前体 FR901379 的产量。
Microb Cell Fact. 2023 Mar 6;22(1):44. doi: 10.1186/s12934-023-02050-0.
4
Overview of Bioactive Fungal Secondary Metabolites: Cytotoxic and Antimicrobial Compounds.生物活性真菌次生代谢产物概述：细胞毒性和抗菌化合物
Antibiotics (Basel). 2022 Nov 11;11(11):1604. doi: 10.3390/antibiotics11111604.
antiSMASH 5.0：二次代谢产物基因组挖掘管道的更新。
Nucleic Acids Res. 2019 Jul 2;47(W1):W81-W87. doi: 10.1093/nar/gkz310.
4
Metabolic engineering of the thermophilic filamentous fungus to produce fumaric acid.嗜热丝状真菌的代谢工程改造以生产富马酸。
Biotechnol Biofuels. 2018 Dec 3;11:323. doi: 10.1186/s13068-018-1319-1. eCollection 2018.
5
Echinocandins in antifungal pharmacotherapy.抗真菌药物治疗中的棘白菌素类药物。
J Pharm Pharmacol. 2017 Dec;69(12):1635-1660. doi: 10.1111/jphp.12780. Epub 2017 Jul 26.
6
Structural diversity in echinocandin biosynthesis: the impact of oxidation steps and approaches toward an evolutionary explanation.棘白菌素生物合成中的结构多样性：氧化步骤的影响及进化解释方法
Z Naturforsch C J Biosci. 2017 Jan 1;72(1-2):1-20. doi: 10.1515/znc-2016-0156.
7
Establishing an efficient gene-targeting system in an itaconic-acid producing Aspergillus terreus strain.在产衣康酸的土曲霉菌株中建立高效的基因靶向系统。
Biotechnol Lett. 2016 Sep;38(9):1603-10. doi: 10.1007/s10529-016-2143-y. Epub 2016 Jun 4.
8
DHN melanin biosynthesis in the plant pathogenic fungus Botrytis cinerea is based on two developmentally regulated key enzyme (PKS)-encoding genes.植物病原真菌灰葡萄孢中DHN黑色素的生物合成基于两个受发育调控的关键酶（聚酮合酶）编码基因。
Mol Microbiol. 2016 Feb;99(4):729-48. doi: 10.1111/mmi.13262. Epub 2015 Nov 24.
9
Genetic transformation of extremophilic fungi Acidea extrema and Acidothrix acidophila.嗜极端真菌Acidea extrema和嗜酸嗜酸丝菌的遗传转化
Folia Microbiol (Praha). 2015 Jul;60(4):365-71. doi: 10.1007/s12223-015-0398-7. Epub 2015 May 2.
10
Cloning, characterization and application of a glyceraldehyde-3-phosphate dehydrogenase promoter from Aspergillus terreus.从土曲霉中克隆、鉴定和应用甘油醛-3-磷酸脱氢酶启动子。
J Ind Microbiol Biotechnol. 2014 Mar;41(3):585-92. doi: 10.1007/s10295-013-1385-0. Epub 2013 Dec 4.