title: Actinobacterial chitinase-like enzymes: profiles of rhizosphere versus non-rhizosphere isolates.
abstract: The objective of this study was to determine if antifungal actinomycetes isolated from rhizosphere and non-rhizosphere soils exhibit different chitinase-like production and (or) induction patterns. Selected isolates from both habitats were compared. Chitinase-like levels and isoform characteristic patterns were evaluated over time in culture fluids of isolates grown on media containing different combinations of colloidal chitin and fungal cell wall (FCW) preparation. Supernatants were also subjected to native and non-native polyacrylamide gel electrophoresis (PAGE), using glycol chitin amended gels. For non-native PAGE, protein samples were denatured by two different approaches. Multiple active bands, ranging from 20 to 53 kDa and present in varying amounts, were detected in gels for most strains. Different substrate preferences were observed among strains, and different chitinase-like enzymes were produced, depending upon the substrate combinations used. The presence of FCW in the medium induced specific chitinase-like enzymes not observed otherwise. Enzymatic activities and profiles of the isolates, however, were strain and substrate specific rather than habitat specific. However, a sagebrush rhizosphere soil had a larger actinomycete community with higher chitinolytic activities than the nearby bulk soil. The use of PAGE to compare chitinase-like proteins induced in media with and without FCW was useful for identifying chitinase-like enzymes potentially involved in antifungal activity.
authors.affiliation: Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow 83844-3052, USA.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Gonzalez-Franco
authors.forename: Lee A
authors.initials: LA
authors.lastname: Deobald
authors.forename: Aaron
authors.initials: A
authors.lastname: Spivak
authors.forename: Don L
authors.initials: DL
authors.lastname: Crawford
mesh_terms: D039903:Actinobacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D019306:Artemisia / Q000382:microbiology; D002473:Cell Wall / Q000378:metabolism; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000737:chemistry / Q000378:metabolism*; D004275:DNA, Ribosomal / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005658:Fungi / Q000737:chemistry; D018517:Plant Roots / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013302:Streptomyces / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D002686:Chitin; D002688:Chitinases