期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 144, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/59152
关键词
Biochemistry; Issue 144; Solid-state NMR; dynamic nuclear polarization (DNP); carbohydrates; cell walls; biomaterials; plant; fungi
资金
- National Science Foundation [NSF OIA-1833040, DMR-1157490]
- State of Florida
- NIH [S10 OD018519]
- NSF [CHE-1229170]
This protocol shows how uniformly C-13, N-15-labeled fungal materials can be produced and how these soft materials should be proceeded for solid-state NMR and sensitivity-enhanced DNP experiments. The sample processing procedure of plant biomass is also detailed. This method allows the measurement of a series of 1D and 2D C-13-C-13/N-15 correlations spectra, which enables high-resolution structural elucidation of complex biomaterials in their native state, with minimal perturbation. The isotope-labeling can be examined by quantifying the intensity in 1D spectra and the polarization transfer efficiency in 2D correlation spectra. The success of dynamic nuclear polarization (DNP) sample preparation can be evaluated by the sensitivity enhancement factor. Further experiments examining the structural aspects of the polysaccharides and proteins will lead to a model of the three-dimensional architecture. These methods can be modified and adapted to investigate a wide range of carbohydrate-rich materials, including the natural cell walls of plants, fungi, algae and bacteria, as well as synthesized or designed carbohydrate polymers and their complex with other molecules.
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