A Novel Assay for Fungal Ketol-isomerase Activity.

• NAKATA MITSUNORI

  Discovery Laboratories, Toyama Chemical Co., Ltd.

• O'ROURKE REBECCA

  MycoLogics, Inc.

• WILSON SHELLY

  MycoLogics, Inc.

• CHILSON KATHERINE

  MycoLogics, Inc.

• SELITRENNIKOFF CLAUDE P.

  MycoLogics, Inc.

抄録

2-Deoxy-D-glucose-6-phosphate ketol-isomerase (EC 2.6.1.16) forms glucosamine-6-phosphate and glutamate from fructose-6-phosphate and glutamine and plays an important role in chitin synthesis in fungi. We have established a new assay for fungal ketol-isomerase activity that is amenable to high throughput screening to identify enzyme inhibitors. Aspergillus fumigatus crude lysate was incubated with substrates and after incubation, reactions were terminated. Glutamate dehydrogenase, nitro blue tetrazolium chloride, phenazine methosulfate and β-NAD were added and the amount of glutamate formed by ketol-isomerase activity was determined by measuring OD_585nm. A feedback inhibitor, UDP-N-acetylglucosamine, of fungal ketol-isomerase was successfully detected by this assay (IC₅₀-0.48 mM). In a pilot scale screening, an active extract from an extremophilic bacterium was found, and the extract showed antifungal activity against A. fumigatus, Candida albicans and C glabrata.

収録刊行物

• The Journal of Antibiotics

  The Journal of Antibiotics 54 (9), 737-743, 2001

  公益財団法人 日本感染症医薬品協会

キーワード

• Drug Discovery
• Pharmacology

詳細情報

• CRID: 1390282679125851648
• NII論文ID: 130003410432
• NII論文ID: 10008470913
• NII書誌ID: AA0069330X
• DOI: 10.7164/antibiotics.54.737
• COI: 1:CAS:528:DC%2BD3MXnt1eksr0%3D
• ISSN: 18811469
• ISSN: 00218820
• PubMed: 11714230
• Web Site: https://search.jamas.or.jp/link/ui/2002102953
• 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