Literature DB >> 34958718

Comprehensive Guide to Extracting and Expressing Fungal Secondary Metabolites with Aspergillus fumigatus as a Case Study.

Grant Nickles1, Isabelle Ludwikoski1, Jin Woo Bok1, Nancy P Keller1,2.   

Abstract

Fungal secondary metabolites (SMs) have captured the interest of natural products researchers in academia and industry for decades. In recent years, the high rediscovery rate of previously characterized metabolites is making it increasingly difficult to uncover novel compounds. Additionally, the vast majority of fungal SMs reside in genetically intractable fungi or are silent under normal laboratory conditions in genetically tractable fungi. The fungal natural products community has broadly overcome these barriers by altering the physical growth conditions of the fungus and heterologous/homologous expression of biosynthetic gene cluster regulators or proteins. The protocols described here summarize vital methodologies needed when researching SM production in fungi. We also summarize the growth conditions, genetic backgrounds, and extraction protocols for every published SM in Aspergillus fumigatus, enabling readers to easily replicate the production of previously characterized SMs. Readers will also be equipped with the tools for developing their own strategy for expressing and extracting SMs from their given fungus or a suitable heterologous model system.
© 2021 Wiley Periodicals LLC. Basic Protocol 1: Making glycerol stocks from spore suspensions Alternate Protocol 1: Creating glycerol stocks from non-sporulating filamentous fungi Basic Protocol 2: Activating spore-suspension glycerol stocks Basic Protocol 3: Extracting secondary metabolites from Aspergillus spp grown on solid medium Alternate Protocol 2: Extracting secondary metabolites from Aspergillus spp using ethyl acetate Alternate Protocol 3: High-volume metabolite extraction using ethyl acetate Alternate Protocol 4: Extracting secondary metabolites from Aspergillus spp in liquid medium Support Protocol: Creating an overlay culture Basic Protocol 4: Extracting DNA from filamentous fungi Basic Protocol 5: Creating a DNA construct with double-joint PCR Alternate Protocol 5: Creating a DNA construct with yeast recombineering Basic Protocol 6: Transformation of Aspergillus spp Basic Protocol 7: Co-culturing fungi and bacteria for extraction of secondary metabolites. © 2021 Wiley Periodicals LLC.

Entities:  

Keywords:  Aspergillus fumigatus; fungi; metabolic extraction; natural products; transformation

Mesh:

Year:  2021        PMID: 34958718      PMCID: PMC8730535          DOI: 10.1002/cpz1.321

Source DB:  PubMed          Journal:  Curr Protoc        ISSN: 2691-1299


  86 in total

1.  CRISPR-Mediated Activation of Biosynthetic Gene Clusters for Bioactive Molecule Discovery in Filamentous Fungi.

Authors:  Indra Roux; Clara Woodcraft; Jinyu Hu; Rebecca Wolters; Cameron L M Gilchrist; Yit-Heng Chooi
Journal:  ACS Synth Biol       Date:  2020-06-26       Impact factor: 5.110

2.  Southern blotting.

Authors:  Ed Southern
Journal:  Nat Protoc       Date:  2006       Impact factor: 13.491

3.  Enhancement of antibiotic and secondary metabolite detection from filamentous fungi by growth on nutritional arrays.

Authors:  G F Bills; G Platas; A Fillola; M R Jiménez; J Collado; F Vicente; J Martín; A González; J Bur-Zimmermann; J R Tormo; F Peláez
Journal:  J Appl Microbiol       Date:  2008-02-19       Impact factor: 3.772

4.  Media and growth conditions for induction of secondary metabolite production.

Authors:  Jens C Frisvad
Journal:  Methods Mol Biol       Date:  2012

5.  Cloning, mapping and molecular analysis of the pyrG (orotidine-5'-phosphate decarboxylase) gene of Aspergillus nidulans.

Authors:  B R Oakley; J E Rinehart; B L Mitchell; C E Oakley; C Carmona; G L Gray; G S May
Journal:  Gene       Date:  1987       Impact factor: 3.688

6.  SMURF: Genomic mapping of fungal secondary metabolite clusters.

Authors:  Nora Khaldi; Fayaz T Seifuddin; Geoff Turner; Daniel Haft; William C Nierman; Kenneth H Wolfe; Natalie D Fedorova
Journal:  Fungal Genet Biol       Date:  2010-06-08       Impact factor: 3.495

7.  Dual-purpose isocyanides produced by Aspergillus fumigatus contribute to cellular copper sufficiency and exhibit antimicrobial activity.

Authors:  Nicholas Raffa; Tae Hyung Won; Andrew Sukowaty; Kathleen Candor; Chengsen Cui; Saayak Halder; Mingji Dai; Julio A Landero-Figueroa; Frank C Schroeder; Nancy P Keller
Journal:  Proc Natl Acad Sci U S A       Date:  2021-02-23       Impact factor: 11.205

8.  Manipulating the yeast genome: deletion, mutation, and tagging by PCR.

Authors:  Jennifer M Gardner; Sue L Jaspersen
Journal:  Methods Mol Biol       Date:  2014

9.  Mating-type factor-specific regulation of the fumagillin/pseurotin secondary metabolite supercluster in Aspergillus fumigatus.

Authors:  Yidong Yu; Adriana Blachowicz; Cornelia Will; Edyta Szewczyk; Steven Glenn; Sabrina Gensberger-Reigl; Minou Nowrousian; Clay C C Wang; Sven Krappmann
Journal:  Mol Microbiol       Date:  2018-10-23       Impact factor: 3.501

10.  Microevolution in the pansecondary metabolome of Aspergillus flavus and its potential macroevolutionary implications for filamentous fungi.

Authors:  Milton T Drott; Tomás A Rush; Tatum R Satterlee; Richard J Giannone; Paul E Abraham; Claudio Greco; Nandhitha Venkatesh; Jeffrey M Skerker; N Louise Glass; Jesse L Labbé; Michael G Milgroom; Nancy P Keller
Journal:  Proc Natl Acad Sci U S A       Date:  2021-05-25       Impact factor: 11.205

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