An Efficient Method for ¹⁵N-Labeling of Chitin in Fungi

Watson, H.R.; Apperley, D.C.; Dixon, D.P.; Edwards, R.; Hodgson, D.R.W.

Abstract

To permit facile 15N solid-state NMR (ssNMR) analysis of the degree of acetylation (DA) of chitinous materials in fungi a method for the introduction of a 15N isotopic label has been developed. Using Penicillium chrysogenum as a model system, a series of 15N-based media were surveyed for their abilities to support mycelial growth, and a rich medium supplemented with (15NH4)2SO4 supported good growth. Uptake of label into chitin extracted from mycelia grown in the rich (15NH4)2SO4-based media was monitored by mass spectrometry, with 1 g/L of (15NH4)2SO4 leading to 65% incorporation. The labeled chitin was studied by ssNMR to determine its DA, and the 15N label permitted measurement of DA to within 0.5% with acquisition times of on the order of half an hour. Similar studies validated the method for DA measurements on chitin from cultures of Aspergillus niger and Mucor rouxii.

Citation

Watson, H., Apperley, D., Dixon, D., Edwards, R., & Hodgson, D. (2009). An Efficient Method for ¹⁵N-Labeling of Chitin in Fungi. Biomacromolecules, 10(4), 793-797. https://doi.org/10.1021/bm8012814

• Journal Article Type: Article
• Publication Date: 2009-02
• Journal: Biomacromolecules
• Print ISSN: 1525-7797
• Electronic ISSN: 1526-4602
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• Volume: 10
• Issue: 4
• Pages: 793-797
• DOI: https://doi.org/10.1021/bm8012814
• Public URL: https://durham-repository.worktribe.com/output/1527848
• Publisher URL: http://pubs.acs.org/doi/abs/10.1021/bm8012814