A unique post-translational processing of an exo-beta-1,3-glucanase of Penicillium sp. KH10 expressed in Aspergillus oryzae.
To characterize an exo-beta-1,3-glucanase (ExgP) of an isolated fungal strain with high laminarin degradation activity, identified as Penicillium sp. KH10, heterologous secretory expression of the ExgP was performed in Aspergillus oryzae. Deduced amino acid sequence of the exgP gene possibly consisted of 989 amino acids which showed high sequence similarity to those of fungal exo-beta-1,3-glucanases belonging to the glycoside hydrolase (GH) family 55. Notably, the purified recombinant ExgP showed a single protein peak in the native state (by gel-permeation chromatographic analysis), but showed two protein bands in the denatured state (by SDS-polyacrylamide gel electrophoresis). These two polypeptides exhibited activity in a coexisting state even under reducing conditions, suggesting that non-covalent association of both polypeptides took place. Taken together with the nucleotide sequence information, the ExgP precursor (104kDa) would be proteolytically processed (cleaved) to generate two protein fragments (42 and 47kDa) and the processed products (polypeptide fragments) would be assembled each other by a non-covalent interaction. Moreover, one of the matured ExgP polypeptides was N-glycosylated by the post-translational modification.
Ooi
Toshihiko
T
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13, W8, Kita-ku, Sapporo 060-8628, Japan. ooi@eng.hokudai.ac.jp
Sato
Hirokazu
H
Matsumoto
Ken'ichiro
K
Taguchi
Seiichi
S
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D043326:Glucan 1,3-beta-Glucosidase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics; D011499:Protein Processing, Post-Translational; D016415:Sequence Alignment; D013379:Substrate Specificity; D013696:Temperature
D005656:Fungal Proteins; D043326:Glucan 1,3-beta-Glucosidase