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Steroid Monooxygenase of Rhodococcus Sequencing of rhodochrous: the Genomic DNA, and Hyperexpression, Purification, and Characterization of the Recombinant Enzyme
https://doi.org/10.24517/00060093
https://doi.org/10.24517/00060093c517976b-2127-4cc8-a8a7-965c059771ff
名前 / ファイル | ライセンス | アクション |
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SC-PR-ITAGAKI-E-126_624.pdf (2.2 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2020-12-03 | |||||
タイトル | ||||||
タイトル | Steroid Monooxygenase of Rhodococcus Sequencing of rhodochrous: the Genomic DNA, and Hyperexpression, Purification, and Characterization of the Recombinant Enzyme | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
ID登録 | ||||||
ID登録 | 10.24517/00060093 | |||||
ID登録タイプ | JaLC | |||||
著者 |
Morii, Shingo
× Morii, Shingo× Sawamoto, Shizue× Yamauchi, Yuhji× Miyamoto, Masahiko× Iwami, Masafumi× Itagaki, Eiji |
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著者別名 |
板垣, 英治
× 板垣, 英治 |
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書誌情報 |
The Journal of Biochemistry 巻 126, 号 3, p. 624-631, 発行日 1999 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 0021-924X | |||||
NCID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA00694073 | |||||
出版者 | ||||||
出版者 | The Japanese Biochemical Society | |||||
出版者(別名) | ||||||
出版者 | 日本生化学会 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Steroid monooxygenase of Rhodococcus rhodochrous is a Baeyer-Villigerase catalyzing the insertion of an oxygen atom between the C17- and C20-carbons of progesterone to produce testosterone acetate. The 5.1-kbp-long BamHI DNA fragment containing the steroid monooxygenase gene, smo, was cloned from the chromosomal DNA and sequenced. The smo gene is 1, 650 nucleotides long, starts with a TTG colon, and ends with a TGA codon. The deduced amino acid sequence indicates that the enzyme protein consist of 549 amino acid residues with a molecular mass of 60, 133. Thus, the molecular mass of the holoenzyme is 60, 919. The amino acid sequence is highly homologous (41.2% identity) to that of cyclohexanone monooxygenase of Acinetobacter sp. In the upstream of the smo gene, the genes of heat shock proteins, dnaK, grpE, and dnaJ, located on the complementary strand, and the DNA-inserts of pSMO and pD1, which contains the ksdD gene, were joined at the BamHI site of the dnaJ gene. The smo gene was modified at the initiation codon to ATG and ligated with an expression vector to construct a plasmid, pSMO-EX, and introduced into Escherichia coli cells. The transformed cells hyperexpressed the steroid monooxygenase as an active and soluble protein at more than 40 times the level in R. rhodochrous cells. Purification of the recombinant monooxygenase from the E. coli cells by simplified procedures yielded about 2.3 mg of enzyme protein/g wet cells. The purified recombinant steroid monooxygenase exhibited indistinguishable molecular and catalytic properties from those of the R. rhodochrous enzyme. | |||||
権利 | ||||||
権利情報 | Copyright © The Japanese Biochemical Society 日本生化学会 | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | http://www.jbsoc.or.jp/ | |||||
関連名称 | http://www.jbsoc.or.jp/ | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://academic.oup.com/jb | |||||
関連名称 | https://academic.oup.com/jb | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://www.jstage.jst.go.jp/browse/biochemistry1922/-char/ja | |||||
関連名称 | https://www.jstage.jst.go.jp/browse/biochemistry1922/-char/ja |