Serial enzymatic hydrolysis of cell walls of two serotypes of yeast-form Histoplasma capsulatum with alpha(1 leads to 3)-glucanase, beta(1 leads to 3)-glucanase, pronase, and chitinase

Abstract

A serial enzymatic hydrolysis procedure for the partial lysis of Histoplasma capsulatum yeast-form cell walls was described, and its application for the differentiation of two serotypes was evaluated. Cell walls were serially digested with alpha(1 leads to 3)-glucanase and beta(1 leads to 3)-glucanse of Cladosporium resinae, then by Pronase, and then by chitinase. The walls of serotype 1, 2, 3 (61.5% digested) were not susceptible to alpha(1 leads to 3)-glucanse, and they contained 30.3% chitin, thus identifying the strain as chemotype 1 (chem 1). Serotype 1, 4 cells walls (51.6% digested) released 27.3% as glucose after treatment with alpha(1 leads to 3)-glucanse and contained 7.8% chitin, compatible with chemotype 2 (chem 2). In addition to quantitating the monomeric products of enzymolysis, I recovered soluble nondialyzable polysaccharide from the digests of both serotypes.