Production of Dihydrofolate Reductase by Cloned<i>Escherichia coli</i>and Its Application to Asymmetric Synthesis of<i>l</i>-Leucovorin

• OHSHIRO Takashi

• KUGE Yukihiro

• IGARASHI Akiko

• MOCHIDA Kenichi

• IWAKURA Masanori

• UWAJIMA Takayuki

書誌事項

タイトル別名

• Production of Dihydrofolate Reductase by Cloned Escherichia coli and Its Application to Asymmetric Synthesis of l-Leucovorin.

抄録

We have investigated culture conditions for production of dihydrofolate reductase by Escherichia coli harboring a high expression plasmid, pTP64-1. Sorbitol addition and pH control were effective for the production of the enzyme in a jar fermentor. The enzyme was purified from a cell-free extract by column chromatographies on DEAE-Cellulofine and Superose Prep12 and showed a single band on SDS-polyacrylamide gel electrophoresis. The reduction of 200 mM dihydrofolate to 6(S)-tetrahydrofolate, an intermediate for l-leucovorin synthesis, was complete in 2 hr under anaerobic conditions, using 1.5 units/ml of the purified enzyme.

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 56 (3), 437-440, 1992

  公益社団法人 日本農芸化学会

キーワード

• Organic Chemistry
• Molecular Biology
• Applied Microbiology and Biotechnology
• General Medicine
• Biochemistry
• Analytical Chemistry
• Biotechnology

詳細情報

• CRID: 1390001206471498496
• NII論文ID: 110002680551
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.56.437
• ISSN: 13476947
• ISSN: 09168451
• PubMed: 1368327
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.56.437
• 本文言語コード: en
• データソース種別:
  • JaLC
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可