Purification and characterization of chitinase A of Streptomyces cyaneus SP-27: an enzyme participates in protoplast formation from Schizophyllum commune mycelia.
A culture filtrate of Bacillus circulans KA-304 grown on a cell-wall preparation of Schizophyllum commune has an activity to form protoplasts from S. commune mycelia. alpha-1,3-Glucanase and chitinase I, which were isolated from the filtrate, did not form the protoplast by itself while a mixture of them showed protoplast-forming activity. Streptomyces cyaneus SP-27 was isolated based on the productivity of chitinase. The culture filtrate of S. cyaneus SP-27 did not form S. commune protoplasts, but addition of it to alpha-1,3-glucanase of B. circulans KA-304 brought about protoplast-forming activity. Chitinase A isolated from the S. cyaneus SP-27 culture filtrate was more effective than chitinase I of B. circulans KA-304 for the protoplast formation in combination with alpha-1,3-glucanase. The N-terminal amino acid sequence of chitinase A (MW 29,000) has a sequential similarity to those of several Streptomycete family 19 chitinases. Chitinase A adsorbed to chitinous substrate and inhibited the growth of Trichoderma reesei mycelia. Anomer analysis of the reaction products also suggested that the enzyme is a family 19 chitinase.
Yano
Shigekazu
S
Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan University, Kusatsu, Shiga, Japan.
Rattanakit
Nopakarn
N
Honda
Arata
A
Noda
Yuta
Y
Wakayama
Mamoru
M
Plikomol
Abhinya
A
Tachiki
Takashi
T
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D001487:Basidiomycota / Q000502:physiology; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004275:DNA, Ribosomal / Q000235:genetics; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism; D025282:Mycelium / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011523:Protoplasts / Q000502:physiology*; D012567:Schizophyllum / Q000201:enzymology / Q000502:physiology*; D013302:Streptomyces / Q000201:enzymology*; D000520:alpha-Glucosidases / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D004275:DNA, Ribosomal; D005656:Fungal Proteins; D002686:Chitin; C042059:glucosidase I; D002688:Chitinases; D000520:alpha-Glucosidases