Structure, heterologous expression, and properties of rice (Oryza sativa L.) family 19 chitinases.
We identified four new family 19 chitinases in Oryza sativa L. cv. Nipponbare: one class I (OsChia1d), two class II (OsChia2a and OsChia2b), and one class IV (OsChia4a). OsChia2a resembled (about 60% identity) the catalytic domains of class I chitinases, but OsChia2b was almost identical (95% identity) to that of the class IV enzyme. OsChia1c, OsChia1c deltaCBD (a deletion of OsChia1c lacking a chitin-binding domain, CBD), and OsChia2b were separately expressed and purified in Pichia pastoris. OsChia1c inhibited fungal growth significantly more than OsChia1c deltaCBD or OsChia2b. The activities of these enzymes on chitin polymers were similar, but they acted differently on N-acetylchitooligosaccharides, (GlcNAC)n. OsChia1c slowly hydrolyzed (GlcNAC)6 and very poorly hydrolyzed (GlcNAC)4 and (GlcNAC)5. In contrast, OsChia2b efficiently hydrolyzed these oligosaccharides. The high antifungal activity and low hydrolytic activity of the class I enzyme towards (GlcNAC)n imply that it participates in the generation of N-acetylchitooligosaccharide elicitors from the cell walls of infecting fungi.
Truong
Nam-Hai
NH
National Food Research Institute, Kannondai 2-1-12, Tsukuba, Ibaraki 305-8642, Japan.
Park
Seung-Moon
SM
Nishizawa
Yoko
Y
Watanabe
Takeshi
T
Sasaki
Takuji
T
Itoh
Yoshifumi
Y
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D001483:Base Sequence; D002384:Catalysis; D002688:Chitinases / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D003470:Culture Media; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005658:Fungi / Q000187:drug effects; D006868:Hydrolysis; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry; D007700:Kinetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology*; D010957:Plasmids / Q000235:genetics; D013045:Species Specificity
D000935:Antifungal Agents; D003470:Culture Media; D018076:DNA, Complementary; D007527:Isoenzymes; D002688:Chitinases