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J Biomed Transl Res. 2017 Sep;18(3):103-107. English. Original Article.
Jung JA , Kim HY , Jo A , Kim JW , Lee WK , Byun JW .
Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.
Foreign Animal Disease Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.
College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Korea.

It has been known that the ability of Shiga toxinproducing Escherichia coli (STEC) to produce Stx2e in culture media plays a role in the diagnosis of edema disease and determination of subunit vaccine candidates in STEC isolates. To examine the efficiency of Stx2e production in several commercial media, a Stx2e-producing strain (KEFS1302) was grown in four different media: ISO-Sensitest broth (ISB), E. coli broth (ECB), trypticase soy broth (TSB), and Mueller Hinton broth (MHB), with or without mitomycin C at 37℃ (250 rpm) for 6 h. Toxin production was measured by enzyme-linked immunosorbent assay. In the presence of mitomycin C, ECB was found to be the most suitable medium, reaching a production peak (OD₆₀₀ = 1.2) at 1 h; Stx2e was mostly produced during the logarithmic phase (within 3 h). On the other hand, toxin production in ISB reached a peak at 3 h after incubation in the absence of mitomycin C. Stx2e was purified by fast protein liquid chromatography (FPLC) using anion-exchange chromatography. The 43 kDa band of Stx2e was confirmed by western blot using the ECB supernatant. Our results showed that ECB and ISB media would be a suitable medium for mass production of Stx2e even if the toxin production is dependent on time.

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