A Neurospora Assay for Pyridoxine

Stokes, J. L.; Larsen, Alma; Woodward, Carl R.; Foster, J. W.

doi:10.1016/s0021-9258(18)51246-1

Cited by 59 publications

(3 citation statements)

References 9 publications

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“…N. crassa was the organism used to generate foundational theories, such as Beadle and Tatum's one‐gene‐one‐enzyme hypothesis (Beadle & Tatum, 1941). N. crassa was also used to study the first conditional mutants (Stokes, Larsen, Woodward, & Foster, 1943). These auxotrophic mutants established the links among genes and enzymes and the ordering of reactions in biosynthetic pathways (McCluskey, Wiest, & Plamann, 2010; Perkins, Radford, & Sachs, 2001; Radford, 2004).…”

Section: Commentarymentioning

confidence: 99%

Targeted Metabolomics Using LC‐MS in Neurospora crassa

et al. 2022

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The filamentous fungus Neurospora crassa has historically been a model for understanding the relationship between genes and metabolism—auxotrophic mutants of N. crassa were used by Beadle and Tatum to develop the one‐gene‐one‐enzyme hypothesis for which they earned the Nobel Prize in 1958. In the ensuing decades, several techniques have been developed for the systematic analysis of metabolites in N. crassa and other fungi. Untargeted and targeted approaches have been used, with a focus on secondary metabolites over primary metabolism. Here, we describe a pipeline for sample preparation, metabolite extraction, Liquid Chromatography‐Mass Spectrometry (LC‐MS), and data analysis that can be used for targeted metabolomics of primary metabolites in N. crassa. Liquid cultures are grown with shaking in a defined minimal medium and then collected using filtration. Samples are lyophilized for 2 days at –80°C, pulverized, and mixed with a solution to extract polar metabolites. The metabolites are separated and identified using LC‐MS, with downstream analysis using Skyline interpretive software. Relative levels of hundreds of metabolites can be detected and compared across strains. © 2022 Wiley Periodicals LLC. Basic Protocol: Metabolite extraction and detection from Neurospora crassa cell cultures using Liquid Chromatography‐Mass Spectrometry

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Section: Commentarymentioning

confidence: 99%

Targeted Metabolomics Using LC‐MS in Neurospora crassa

et al. 2022

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“…An interesting practical application of mutant strains of Neurospora is in bioassays for particular vitamins and amino acids. So far, procedures have been developed for bioassays for pyridoxin (299,300), p-aminobenzoic acid (312,317), choline (136, 189a), inositol (15), and leucine (240,255). An advantage of using mutant strains over naturally occurring organisms is that there is a greater freedom of choice as to the organism used and also as to its specificity.…”

Section: N Biosynthetic Processes In Neurosporamentioning

confidence: 99%

Biochemical Genetics.

Beadle¹

1945

Chem. Rev.

233

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“…We are indebted to Mr. Frank Streightoff who carried out these assays according to the method of Stokes, et al(4) using the organism Neurospora sitophila; Pyridoxineless No. 9276 of the American Type Culture Collection.…”

mentioning

confidence: 99%

Attempts to Rearrange 3,4-Substituted-2-Acetylfurans to Pyridoxine

Kornfeld¹

1955

J. Org. Chem.

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A Neurospora Assay for Pyridoxine

Cited by 59 publications

References 9 publications

Targeted Metabolomics Using LC‐MS in Neurospora crassa

Targeted Metabolomics Using LC‐MS in Neurospora crassa

Biochemical Genetics.

Attempts to Rearrange 3,4-Substituted-2-Acetylfurans to Pyridoxine

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