Characterization of nutrient status of Halamphora luciae (Bacillariophyceae) using matrix-assisted ultraviolet laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)

Abstract

The effects of N and P depletion on the production and structural characterization of the cellular carbohydrate polymers of the estuarine diatom Halamphora luciae in batch culture were examined using matrix-assisted laser desorption-ionization time-of flight mass spectrometry (MALDI-TOF MS) complemented with monosaccharide composition determination and structural analyses by methylation of aqueous extracted product. The MALDI MS analysis of the cells showed a similar profile in control and N- and P-depleted media, with a displacement to higher molecular weight for cells grown in depleted media. In the monosaccharide analyses, both nutrient depletion and culture ageing led to an increase in glucose content, indicating that MALDI-TOF MS in whole cells was detecting the changes in chrysolaminarin. The maxima for the ions from f/2-P and to a lesser extent in f/2-N were displaced to higher m/z values indicating a higher degree of polymerization (DP). Methylation analysis confirmed the presence of chrysolaminarin, a (1→3)-β-d-glucan with branching in C2 and C6, where the glucan backbone had a substitution every four glucose residues. The (1→3)-β-d-glucan was also detected in the cingule by fluorescence with aniline blue.