J Bacteriol Virol.  2009 Dec;39(4):269-276. 10.4167/jbv.2009.39.4.269.

Bacteriocin from Purple Nonsulfur Phototrophic Bacteria, Rhodobacter capsulatus

• Lee SS
• Oh TJ
• Kim J
• Kim JB
• Lee HS

• KMID: 2168538
• DOI: http://doi.org/10.4167/jbv.2009.39.4.269

Abstract

To find whether productivity of bacteriocin is controlled between different species under unusual cultural conditions, we used Rhodobacter capsulatus ATCC 17016 as a producer and Rhodopseudomonas palustris ATCC 17003 as an indicator. Rhodobacter capsulatus was cultured under aerobic conditions in the dark in Lascelles medium containing 0.3% Triton X-100. As a result, bacteriocin productivity increased enormously. The optimal pH range of bacteriocin production was 6~7.8. Through partial purification of bacteriocin, the molecular weight was roughly estimated at 14 kDa. Plasmid had no influence on bacteriocin production by Rhodobacter capsulatus. Our findings indicate that culture conditions affect bacteriocin productivity between more distantly related species, and bacteriocin of Rhodobacter capsulatus is not encoded by a plasmid.

Keyword

Bacteriocin; Rhodobacter capsulatus; Lascelles medium

MeSH Terms

Bacteria
Efficiency
Hydrogen-Ion Concentration
Molecular Weight
Octoxynol
Plasmids
Rhodobacter
Rhodobacter capsulatus
Rhodopseudomonas
Octoxynol

Figure

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  Figure 1. Cell-bound bacteriocin inhibition of Rhodopseudomonas palustris ATCC 17003 (indicator) lawn on Lascells medium by Rhodobacter capsulatus ATCC 17016 (producer).

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  Figure 2. The growth rate (OD) of Rhodobacter capsulatus ATCC 17016 growing under dark aerobic conditions at various pH.

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  Figure 3. Mono Q anion-exchange chromatography on FPLC. Crude cell-bound bactoriocin (25 mg/ml × 0.5 ml) was applied to Mono Q/HR anion-exchange column (5 × 5 cm) on FPLC equilibrated with 20 mM-ethanolamine/HCl (pH 8.5). Bacteriocin was eluted by a linear NaCl gradient (0~1 M) for 28 min at a flow rate of 2 ml/min per at 280 nm. Note: (—), protein; (–), NaCl gradient; (↓), anti-microbial activity of bacteriocin.

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  Figure 4. Bacteriocin inhibition of Rhodopseudomonas palustris ATCC 17003 indicator lawn on Lascells medium by fraction 11.

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  Figure 5. Polycolonal antibody dilution curve of bacteriocin in indirect ELISA

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  Figure 6. Agarose gel electrophoresis of plasmid DNA from purple nonsulfur photosynthetic bacteria grown in light anaerobic culture (A) and dark aerobic culture (B). Note: lane A: Rhodobacter capsulatus ATCC 17016 (R. sphaeroides ATCC 17023 in B), lane B: Rhodobacter capsulatus DSM 152 (DSM 1710 in B), lane C: Rhodobacter capsulatus DSM 938, lane D: λ DNA digested Hind III (23.1, 9.4, 6.5, 4.3, 2.3, 2.0 kb in order), lane E: Rhodobacter capsulatus DSM 1710 (DSM 152 in B), lane F: Rhodobacter sphaeroides ATCC 17023 (R. capsulatus DSM 17016 in B)

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  Figure 7. Agarose gel electrophoresis of plasmid DNA from Rhodobacter capsulatus ATCC 17016 treated with ethidium bromide (A) and Rhodobacter sphaeroides ATCC 17023 treated with mitomycin C (B) grown in the Lascelles medium. Note for Fig. 7A: lane A to D: cured cell with 4.0, 2.0, 1.0, 0.5 μg/ml ethidium bromide, respectively; lane E: λ DNA digested Hind III and pst I. Note for Fig. 7B: lane A: λ DNA digested Hind III and pst I; lane B & C: cured cell with 20 μg/ml mitocycin C; lane D & E: cured cell with 15 μg/ml mitocycin C.

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