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http://dx.doi.org/10.5352/JLS.2002.12.4.483

Purification and Characterization of an Insect Antibacterial Peptide, Defensin, Expressed in Saccharomyces cerevisiae  

강대욱 (한국생명공학연구원)
이준원 (한국생명공학연구원)
김보연 (한국생명공학연구원)
안종석 (한국생명공학연구원)
Publication Information
Journal of Life Science / v.12, no.4, 2002 , pp. 483-489 More about this Journal
Abstract
We investigated the biochemical properties of insect defensin expressed and secreted from Saccharomyces corevisiae. The defensin showed extremely high resistance to boiling for up to 30 min and to pH values tested from 2.0 to 12.0. The treatment of defensin with various proteases abolished antibacterial activity. However, amylases, cellulase, lipase and catalase had no effect on the activity. The defensin was purified to homogeneity through ammonium sulfate concentration of culture supernatant, SP-Sepharose column chromatography and RP-HPLC. Tricin-SDS-PAGE analysis revealed that the molecular weight of the defensin was about 4.0 kDa. The antibacterial activity of the purified defensin was verified by renaturation of stained gel and gel pouring assay using Micrococcus luteus as a test organism.
Keywords
Defensin; Saccharomyces cerevisine; purification; SP-Sepharose chromatography; HPLC; heat resistance; pH resistance;
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