『徳島大学 教育・研究者情報データベース (EDB)』---[学外] /
ID: Pass:

登録内容 (EID=63979)

EID=63979EID:63979, Map:0, LastModified:2018年12月3日(月) 14:35:20, Operator:[大家 隆弘], Avail:TRUE, Censor:0, Owner:[佐藤 高則], Read:継承, Write:継承, Delete:継承.
種別 (必須): 学術論文 (審査論文) [継承]
言語 (必須): 英語 [継承]
招待 (推奨):
審査 (推奨): Peer Review [継承]
カテゴリ (推奨): 研究 [継承]
共著種別 (推奨):
学究種別 (推奨):
組織 (推奨): 1.青山学院大学.理工学部 [継承]
2.Academy of Russia [継承]
著者 (必須): 1.佐藤 高則
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
2. (英) Samejima Tatsuya (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
3. (英) Watanabe Machiko (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
4. (英) Nogi Shin-ichi (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
5. (英) Takahashi Yoshimasa (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
6. (英) Kaji Hiroyuki (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
7. (英) Teplyakov Alexei (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
8. (英) Obmolova Galya (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
9. (英) Kuranova Inna (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
10. (英) Ishii Keisuke (日) (読)
役割 (任意): 共著 [継承]
貢献度 (任意):
学籍番号 (推奨):
[継承]
題名 (必須): (英) Molecular Cloning, Expression and Site-Directed Mutagenesis of Inorganic Pyrophosphatase from Thermus thermophilus HB8  (日)    [継承]
副題 (任意):
要約 (任意): (英) The genomic DNA encoding the inorganic pyrophosphatase from an extremely thermophilic bacterium, Thermus thermophilus HB8 (ATCC27634), was isolated by colony hybridization with a probe designed as a part of gene amplified by the PCR method, which was derived from the partial amino acid sequence of the enzyme. The DNA was cloned into a plasmid vector, pUC118, after digestion with BamHI. The inserted nucleotide fragment was about 1.8 kbp in length and the nucleotide sequence included a 525 bp open reading frame. The deduced amino acid sequence was completely identical with that of the enzyme determined by automated Edman analysis of peptide fragments isolated from digests obtained with Staphylococcus aureus V8 protease and Achromobacter protease I, and also from products obtained on chemical cleavage with cyanogen bromide and 70% formic acid. The subunit of this enzyme is composed of 174 amino acid residues with a calculated molecular weight of 19,084. Then, the gene was overexpressed in Escherichia coli BL21 (DE3) using a plasmid vector, pET15b, system. The recombinant enzyme was fully active, and exhibited higher thermostability than the E. coli enzyme. Amino acid residues located on the surface of the recombinant enzyme were determined by means of limited proteolysis, and the results revealed that the environment of Lys residues is almost the same as the crystal structure reported previously [Teplyakov, A. et al. (1994) Protein Sci. 3, 1098-1107]. Furthermore, the roles of two tryptophan residues were investigated by site-directed mutagenesis, which indicated that they may be responsible for the structural integrity and thermostability.  (日) 高度好熱性細菌Thermus thermophilusより,無機ピロリン酸加水分解酵素遺伝子のクロ-ニングを行ない,その全一次構造を決定した. また,大腸菌により発現させた組み換え体酵素は,十分な酵素活性および耐熱性を示した.さらに,プロテア-ゼ限定分解による溶液構造の推定や,部位特異的変異法を用いて作製した変異体の解析から,本酵素の耐熱化に寄与する部位について考察を行なった.   [継承]
キーワード (推奨): 1. (英) Amino Acid Sequence (日) (読) [継承]
2. (英) Amino Acid Substitution (日) (読) [継承]
3. (英) Base Sequence (日) (読) [継承]
4. (英) Cloning, Molecular (日) (読) [継承]
5. (英) DNA, Bacterial (日) (読) [継承]
6. (英) Enzyme Stability (日) (読) [継承]
7. (英) Escherichia coli (日) (読) [継承]
8. (英) Hydrolysis (日) (読) [継承]
9. (英) Inorganic Pyrophosphatase (日) (読) [継承]
10. (英) Molecular Sequence Data (日) (読) [継承]
11. (英) Mutagenesis, Site-Directed (日) (読) [継承]
12. (英) Pyrophosphatases (日) (読) [継承]
13. (英) Recombinant Proteins (日) (読) [継承]
14. (英) Sequence Homology, Amino Acid (日) (読) [継承]
15. (英) Thermus thermophilus (日) (読) [継承]
16. (英) Tryptophan (日) (読) [継承]
発行所 (推奨): 日本生化学会 [継承]
誌名 (必須): The Journal of Biochemistry ([日本生化学会])
(pISSN: 0021-924X, eISSN: 1756-2651)

ISSN (任意): 0021-924X
ISSN: 0021-924X (pISSN: 0021-924X, eISSN: 1756-2651)
Title: Journal of biochemistry
Title(ISO): J Biochem
Supplier: Oxford University Press
Publisher: Oxford University Press
 (NLM Catalog  (医中誌Web  (J-STAGE  (Scopus  (CrossRef (Scopus information is found. [need login])
[継承]
[継承]
(必須): 124 [継承]
(必須): 1 [継承]
(必須): 79 88 [継承]
都市 (任意):
年月日 (必須): 西暦 1998年 1月 1日 (平成 10年 1月 1日) [継承]
URL (任意):
DOI (任意):
PMID (任意): 9644249    (→Scopusで検索) [継承]
NAID (任意):
WOS (任意): 000074726400013 [継承]
Scopus (任意):
評価値 (任意):
被引用数 (任意):
指導教員 (推奨):
備考 (任意): 1.(英) Article.DataBankList.DataBank.DataBankName: GENBANK  (日)    [継承]
2.(英) Article.DataBankList.DataBank.AccessionNumberList.AccessionNumber: AB010580  (日)    [継承]
3.(英) Article.DataBankList.DataBank.AccessionNumberList.AccessionNumber: P38576  (日)    [継承]
4.(英) Article.PublicationTypeList.PublicationType: Journal Article  (日)    [継承]
5.(英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't  (日)    [継承]

標準的な表示

和文冊子 ● Takanori Satoh, Tatsuya Samejima, Machiko Watanabe, Shin-ichi Nogi, Yoshimasa Takahashi, Hiroyuki Kaji, Alexei Teplyakov, Galya Obmolova, Inna Kuranova and Keisuke Ishii : Molecular Cloning, Expression and Site-Directed Mutagenesis of Inorganic Pyrophosphatase from Thermus thermophilus HB8, The Journal of Biochemistry, Vol.124, No.1, 79-88, 1998.
欧文冊子 ● Takanori Satoh, Tatsuya Samejima, Machiko Watanabe, Shin-ichi Nogi, Yoshimasa Takahashi, Hiroyuki Kaji, Alexei Teplyakov, Galya Obmolova, Inna Kuranova and Keisuke Ishii : Molecular Cloning, Expression and Site-Directed Mutagenesis of Inorganic Pyrophosphatase from Thermus thermophilus HB8, The Journal of Biochemistry, Vol.124, No.1, 79-88, 1998.

関連情報

Number of session users = 0, LA = 0.83, Max(EID) = 374067, Max(EOID) = 1001448.