Mycosubtilins B and C were isolated from the culture medium of Bacillus subtilis. The acid hydrolysates of these new antifungal antibiotics, like mycosubtilin, contain alpha-amino acids (Asp3, Glu1, Pro1, Ser1 and Tyr1) and a mixture of iso-C16, n-C16, iso-C17 and anteiso-C17 beta-amino acids. Mycosubtilins B and C differ by the presence of a carboxyl group and of a carboxymethyl group, respectively, instead of a carboxamide group in previously described mycosubtilin.