@techreport{oai:soar-ir.repo.nii.ac.jp:00014469,
 author = {下坂,  誠 and 小平,  律子},
 month = {Mar},
 note = {A gene (csnC) Coding for chitosanase secreted in medium was isolated from the koji mold Aspergillus oryzae. CsnC ORF contained serine- and threonine-rich linker and three repeated sequences in its C-terminal, which were not observed in other fungal chitosanases. Tne cDNA corresponding to mature chitosanase was synthesized by a reverse transcriptase-PCR, and introduced into cells of the yeast Pichia pastoris by using an expression vector The resulting transformant secreted active recombinant chitosanase. A protein lacking the repeated sequence (CsnCΔR3) was also expressed and used for kinetic analysis. The repeated sequence was found to be involved in binding of the enzyme to insoluble substrate chitosan. Environmental DNA was prepared from microorganisms in soil amended with chitin. A part of 16S ribosomal DNA was PCR-amplified by using the environmental DNA as a template. The resulting DNA fragments were fractionated by a denaturant gradient gel electrophoresis (DGGE). The intensities of several DNA bands increased significantly after addition of chitin. Determination of nucleotide sequence of these DNA fragments indicated that most of them showed a similarity, to 16S rDNA sequences derived from unculturable bacteria deposited in the database.},
 title = {15-2-8 : キチン･キトサンの酵素変換による高機能化に関する研究},
 year = {2005}
}