Isolation and Characterization of an Extracellular Antimicrobial Protein from Aspergillus oryzae

A 17 kDa antimicrobial protein was isolated from growth medium containing the filamentous fungus Aspergillus oryzae by extracting the supernatants from the culture media, ion exchange chromatography on CM-sepharose, and C18 reverse-phase high-performance liquid chromatography. This antimicrobial pro...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2008-10, Vol.56 (20), p.9647-9652
Main Authors: Park, Seong-Cheol, Yoo, Nae Choon, Kim, Jin-Young, Park, Hae Kyun, Chae, Byung Jo, Shin, Song Yub, Cheong, Hyeonsook, Park, Yoonkyung, Hahm, Kyung-Soo
Format: Article
Language:English
Subjects:
Amino Acid Sequence
amino acid sequences
Anti-Bacterial Agents - chemistry
Anti-Bacterial Agents - isolation & purification
Anti-Bacterial Agents - metabolism
Anti-Bacterial Agents - pharmacology
anti-infective agents
Antifungal Agents - chemistry
Antifungal Agents - isolation & purification
Antifungal Agents - metabolism
Antifungal Agents - pharmacology
Antimicrobial activity
antimicrobial properties
antimicrobial proteins
Aspergillus oryzae
Aspergillus oryzae - chemistry
Aspergillus oryzae - metabolism
Bacteria - drug effects
bioactive properties
Biological and medical sciences
carbon
cell culture
Colletotrichum coccodes
Escherichia coli O157:H7
exAP-AO17
Extracellular Space - chemistry
Extracellular Space - metabolism
Food Chemistry/Biochemistry
Food industries
Food microbiology
Fundamental and applied biological sciences. Psychology
fungal proteins
Fungal Proteins - chemistry
Fungal Proteins - isolation & purification
Fungal Proteins - metabolism
Fungal Proteins - pharmacology
Fungi - drug effects
Gibberella circinata
heat stress
hemolytic activity
homology with a collagen
Humans
Molecular Sequence Data
nitrogen
pathogens
physiological regulation
protein synthesis
secretion
Sequence Alignment
sodium chloride
Staphylococcus aureus
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Description
Summary:A 17 kDa antimicrobial protein was isolated from growth medium containing the filamentous fungus Aspergillus oryzae by extracting the supernatants from the culture media, ion exchange chromatography on CM-sepharose, and C18 reverse-phase high-performance liquid chromatography. This antimicrobial protein, which we considered to be an extracellular antimicrobial protein from A. oryzae (exAP-AO17), possessed antimicrobial activity but lacked hemolytic activity. The exAP-AO17 protein strongly inhibited pathogenic microbial strains, including pathogenic fungi, Fusarium moniliform var. subglutinans and Colletotrichum coccodes, and showed antibacterial activity against bacteria, including E. coli O157 and Staphylococcus aureus. To confirm that the protein acts as a regulation factor for extracellular secretion, we examined growth under varying conditions of N sources, C sources, ions, ambient pH, and stress. Various culture conditions were found to induce characteristic changes in the expression of protein synthesis as analyzed by sodium dodecyl sulfate−polyacrylamide gel electrophoresis. Highly basic polypeptides were regulated by suppressing the ambient pH under acidic conditions and strongly induced under alkaline conditions, thus confirming that pH regulation is physiologically relevant. The expression of exAP-AO17 was upregulated by heat shock upon growth in the presence of NaCl. Automated Edman degradation showed that the N-terminal sequence of exAP-AO17 was NH2-GLPGPAGAVGFAGKDQNM−. ExAP-AO17 showed partial sequence homology with a collagen belonging to the animal source. These results suggest that exAP-AO17 is an excellent candidate as a lead compound for the development of novel oral or other types of anti-infective agents.
ISSN:0021-8561
1520-5118
DOI:10.1021/jf802373h