J Biomed Transl Res.  2017 Sep;18(3):103-107. 10.12729/jbtr.2017.18.3.103.

Enrichment media for Stx2e production in Shigatoxin-producing Escherichia coli

• Jung JA
• Kim HY
• Jo A
• Kim JW
• Lee WK
• Byun JW

• KMID: 2394595
• DOI: http://doi.org/10.12729/jbtr.2017.18.3.103

Abstract

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.

Keyword

edema disease; mitomycin C; Stx2e; Shiga toxin-producing Escherichia coli (STEC); E. coli broth medium

MeSH Terms

Blotting, Western
Chromatography
Chromatography, Liquid
Culture Media
Diagnosis
Edema
Enzyme-Linked Immunosorbent Assay
Escherichia coli*
Escherichia*
Hand
Mitomycin
Shiga-Toxigenic Escherichia coli
Culture Media
Mitomycin