Cyclo(L-leucyl-L-prolyl) produced by Achromobacter xylosoxidans inhibits aflatoxin production by Aspergillus parasiticus

• Published in: Applied and Environmental Microbiology 2004-12, Vol.70 (12), p.7466-7473
• Main Authors: Yan, P.S, Song, Y, Sakuno, E, Nakajima, H, Nakagawa, H, Yabe, K
• Format: Article
• Language: English
• Subjects: Achromobacter denitrificans - classification; Achromobacter denitrificans - genetics; Achromobacter denitrificans - growth & development; Achromobacter denitrificans - metabolism; Achromobacter xylosoxidans; aflatoxins; Aflatoxins - antagonists & inhibitors; Aflatoxins - biosynthesis; amino acid derivatives; Anthraquinones - metabolism; Aspergillus - drug effects; Aspergillus - metabolism; Aspergillus flavus; Aspergillus parasiticus; Bacteria; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; biosynthesis; Biotechnology; Carcinogens; Culture Media; cyclic peptides; Fundamental and applied biological sciences. Psychology; Fungal Proteins - genetics; Fungal Proteins - metabolism; Fungi; gene expression; genes; messenger RNA; metabolic inhibitors; Microbiology; Miscellaneous; Mission oriented research; Molecular Sequence Data; Mycology; nucleotide sequences; Parasites; Peptides, Cyclic - biosynthesis; Peptides, Cyclic - chemistry; Peptides, Cyclic - isolation & purification; Peptides, Cyclic - pharmacology; ribosomal DNA; ribosomal RNA; RNA, Ribosomal, 16S - genetics; Sequence Analysis, DNA; Toxins; transcription (genetics)
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.70.12.7466-7473.2004

Aflatoxins are potent carcinogenic and toxic substances that are produced primarily by Aspergillus flavus and Aspergillus parasiticus. We found that a bacterium remarkably inhibited production of norsolorinic acid, a precursor of aflatoxin, by A. parasiticus. This bacterium was identified as Achromobacter xylosoxidans based on its 16S ribosomal DNA sequence and was designated A. xylosoxidans NFRI-A1. A. xylosoxidans strains commonly showed similar inhibition. The inhibitory substance(s) was excreted into the medium and was stable after heat, acid, or alkaline treatment. Although the bacterium appeared to produce several inhibitory substances, we finally succeeded in purifying a major inhibitory substance from the culture medium using Diaion HP20 column chromatography, thin-layer chromatography, and high-performance liquid chromatography. The purified inhibitory substance was identified as cyclo(L-leucyl-L-prolyl) based on physicochemical methods. The 50% inhibitory concentration for aflatoxin production by A. parasiticus SYS-4 (= NRRL2999) was 0.20 mg ml(-1), as determined by the tip culture method. High concentrations (more than 6.0 mg ml(-1)) of cyclo(L-leucyl-L-prolyl) further inhibited fungal growth. Similar inhibitory activities were observed with cyclo(D-leucyl-D-prolyl) and cyclo(L-valyl-L-prolyl), whereas cyclo(D-prolyl-L-leucyl) and cyclo(L-prolyl-D-leucyl) showed weaker activities. Reverse transcription-PCR analyses showed that cyclo(L-leucyl-L-prolyl) repressed transcription of the aflatoxin-related genes aflR, hexB, pksL1, and dmtA. This is the first report of a cyclodipeptide that affects aflatoxin production.