Two contrasting cyanobacterial species (Anabaena fertilissima and Anabaena sphaerica) were selected based on differences in antifungal behavior in order to study the mechanism for production of an antifungal enzyme and the genes responsible for this production. In A. fertilissima, chitosanase and antifungal activities were increased significantly under of growth-limiting conditions (8 of light and 16 h of darkness). The lack of such activities in A. sphaerica was associated with high levels of protein that accumulated during the stationary phase (at 28 days) under the same light conditions. The gene putatively responsible for chitosanase and antifungal activities was amplified using specific primers, and sequence analysis of the amplified products (1.086 and 1.101 kb in A. sphaerica and A. fertilissima, respectively) showed that they belong to the glycoside hydrolase 3 ( Cyanobacteria comprise a heterogeneous assemblage of photosynthetic prokaryotes having extraordinary biosynthetic potential and a repertoire of diverse metabolic activities. They are an important source of novel antifungal, antibacterial, and herbicidal or weedicidal compounds, which have been implicated in allelopathic interactions in water and soil (29). A majority of these metabolites are biologically active and are products of either nonribosomal polypeptide (NRP) or mixed polyketide-NRP biosynthetic pathways. The toxins produced by cyanobacteria are greatly influenced by various physiological and environmental factors, including light, temperature, nutrients, and pH (5).The antifungal properties of cyanobacterial metabolites, most of which have not been exploited, have immense potential in agriculture for use against fungal plant pathogens. In bacteria, lytic enzymes, such as chitinases, chitosanases, proteases, and -1,3-glucanases, are known to have key roles in biocontrol of various soilborne fungal pathogens. Chitin is a linear polymer of 1,4-linked N-acetyl-D-glucosamine (GlcNAc) residues, and the deacetylated derivative of chitin is chitosan. In contrast to chitin, chitosan has been found in very few organisms, but it has been found in members of the Zygomycota, such as Mucor rouxii (42), Absidia coerulea (22), and
Rhizopus oryzae (8). The filamentous fungi Lentinus edodes andPleurotus sajo-caju were investigated to determine their abilities to produce chitosan (30). Chitosan and chitosan-glucan complexes have been found in the mycelia of Aspergillus niger, Humicola lutea, and Fusarium moniliforme (39). Chitosanases (EC 3.2.1.132) produced by bacteria are classified into five glycoside hydrolase (GH) families (families 5, 8, 46, 75, and 80) (6, 10, 11, 12, 13). Families 5 and 8 are composed of enzymes that are hydrolytic with glycosides, and the family 46, 75, and 80 enzymes studied so far are chitosanases. This classification of chitosanases is based on amino acid sequence similarities of the catalytic domains. Recently, the family 46 chitosanase of Amycolatopsis sp. CsO-2 responsible for antifungal activity against Rhizopus oryzae ...