海洋環境からのコレステロールエステラーゼ生産放線菌の分離と諸性状("Isolation and Characterization of Cholesterol Esterase-Producing Actinomycetes from the Mmarine Environment")
- 標題
- 海洋環境からのコレステロールエステラーゼ生産放線菌の分離と諸性状("Isolation and Characterization of Cholesterol Esterase-Producing Actinomycetes from the Mmarine Environment")
- 作者
- 今田千秋・小野寺かおる・寺原 猛・小林武志・山田勝久(Chiaki Imada, Kaoru Onodera, Takeshi Terahara, Takeshi Kobayashi and Katsuhisa Yamada)
- 文件屬性
- 日本研究
- 知識分類
- 水產養殖
- 出版年
- 2014
- 刊名
- 海洋深層水研究
- 卷
- 14
- 期
- 3
- 頁
- P167-176
- 點閱數
- 2437
摘要
日本列島の岩手県から沖縄県および東シナ海の海底堆積物・生物・海浜砂試料から27セで分離培 養した放線菌合計611 株のコレステロールエステラーゼ（CHE) 生産菌を探索し，幻0 株の陽性株 を得，ヒット率（供試菌株数に対する陽性菌数の割合）は49.1％であった． 最高ヒット率海域は千 葉県館山湾で743% (35株中26株）. 分離サンプルでは，海岸砂のヒット率が67.8% (90株中61 株）と最高であった． これら陽性300 株中，CHE 高活性3株（いずれも海底堆積物）が得られた； 東京湾（2009年5 月，水深5m,A株と命名），東京湾（2009年11 月，水深20m, B株と命名）, 東シナ海（2009 年10 月，水深300m, C 株と命名）. これら3 株の1田rDNA の塩基配列解析から， A,B株はともにStrゆtomyces rubrolavendulae，またC株はS. glauc加如rと高い相同性を示した．
A total of 611 cholesterol esterase (CHE) producing actinomycete strains were isolated and characterized from marine bottom sediments, marine organisms and beach sands collected from bays coastal and offshore waters and beaches facing the Pacific and the East China Sea around Japan. Three hundred positive strains exhibiting CHE activity were isolated. The percentage of positive strain (hit rate) was 49.1%. Highest hit rate of 74.3%(26 positive strains out of 35 strains) was observed in Tateyama Bay in Chiba Prefecture. Among the various samples, the highest hit rate of 67.8%(61 positive strains out of 90 strains) was observed in the samples from coastal sand. Among the 300 positive strains, 3 strains which had high CHE activity were selected and used for further studies. Two strains named A and B were isolated from sediment samples from Tokyo Bay in May (5 m) and October 2009 (20 m), respectively, whereas one strain named C was isolated from the sediment sample obtained from the East China Sea (300 m). From the 16S rDNA analysis strains A and B showed high homology with Strゆtomyces rubrolavendulae whereas strain C was close to S. glaucin如r. Among the three, strain B showed stable highest CHE activity and was selected for the purification and characterization of the enzyme. The CHE was isolated from the culture supernatant of strain B by ammonium sulfate precipitation, anion-exchange chromatography and gel filtration. The final preparation showed a single band on native electrophoresis. The molecular weight of the purified CHE was 622 kDa. The CHE exists as dimer forms composed of 22.7 kDa and 39.5 kDa in water solution. The CHE was stable up to 70セ and also showed pH stability from 4 to 10. The enzyme decomposed substrates such as linoleic acid cholesterol and oleic acid cholesterol.
A total of 611 cholesterol esterase (CHE) producing actinomycete strains were isolated and characterized from marine bottom sediments, marine organisms and beach sands collected from bays coastal and offshore waters and beaches facing the Pacific and the East China Sea around Japan. Three hundred positive strains exhibiting CHE activity were isolated. The percentage of positive strain (hit rate) was 49.1%. Highest hit rate of 74.3%(26 positive strains out of 35 strains) was observed in Tateyama Bay in Chiba Prefecture. Among the various samples, the highest hit rate of 67.8%(61 positive strains out of 90 strains) was observed in the samples from coastal sand. Among the 300 positive strains, 3 strains which had high CHE activity were selected and used for further studies. Two strains named A and B were isolated from sediment samples from Tokyo Bay in May (5 m) and October 2009 (20 m), respectively, whereas one strain named C was isolated from the sediment sample obtained from the East China Sea (300 m). From the 16S rDNA analysis strains A and B showed high homology with Strゆtomyces rubrolavendulae whereas strain C was close to S. glaucin如r. Among the three, strain B showed stable highest CHE activity and was selected for the purification and characterization of the enzyme. The CHE was isolated from the culture supernatant of strain B by ammonium sulfate precipitation, anion-exchange chromatography and gel filtration. The final preparation showed a single band on native electrophoresis. The molecular weight of the purified CHE was 622 kDa. The CHE exists as dimer forms composed of 22.7 kDa and 39.5 kDa in water solution. The CHE was stable up to 70セ and also showed pH stability from 4 to 10. The enzyme decomposed substrates such as linoleic acid cholesterol and oleic acid cholesterol.