Analysis of multi‐domain cellulolytic enzymes by UV‐laser induced desorption mass spectrometry

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Abstract
This paper reports the application of matrix‐assisted UV‐laser induced desorption (LID) mass spectrometry for the analysis of some extracellular multi‐domain cellulolytic enzymes, in the molecular weight range 5000–90 000 u, produced by the fungi Phanerochaete chrysosporium and Trichoderma reesei. A high sensitivity (1–10pmol of protein), has been attained in obtaining the mass spectra of these highly glycosylated enzymes and some of their peptides, with carbohydrate content in the 10–40% range. The desorption of samples containing the enzymes mixed with a protein having an accurately known molecular weight—an internal calibrant—is also demonstrated. Spectra from complete enzymes and their functional domains are presented to explore the possibilities of employing LID for obtaining structural information.