Biochemical and Molecular Characterization of
            <i>Obesumbacterium proteus</i>
            , a Common Contaminant of Brewing Yeasts

Prest, Andrew G.; Hammond, J. R.; Stewart, Gordon S. A. B.

doi:10.1128/aem.60.5.1635-1640.1994

“…Automated ribotyping with both EcoRI (first choice in the system) and PvuII restriction enzymes was shown to differentiate O. proteus biotypes 1 and 2 and H. alvei, which supported the results of Prest et al (1994) obtained using manual ribotyping with EcoRI enzyme. Different restriction enzymes have different discrimination powers, as cleavage results in different sets of DNA fragments and different riboprints.…”

Section: Discussionsupporting

confidence: 67%

Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1

Koivula

¹

,

Juvonen

²

,

Haikara

³

et al. 2006

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Aims: To study the ability of automated ribotyping to characterize Obesumbacterium proteus and Hafnia alvei, to design primers and to evaluate standard end‐point and real‐time PCR for the detection of O. proteus biotype 1 in beer and in brewers's yeast‐containing samples. Methods and Results: Automated ribotyping was carried out using the standard method with EcoRI and PvuII. The digestions with both enzymes clearly differentiated O. proteus biotypes 1 and 2 and H. alvei. PCR primers were designed according to the 16S rRNA gene sequence of the O. proteus type strain. Two primer sets (Obs137–Obs558 and Obs137–Obs617) detected O. proteus biotype 1 and H. alvei but not O. proteus biotype 2 or other tested beer spoilage bacteria (40 species) in the end‐point and real‐time PCR, indicating their high specificity. The detection limit for O. proteus was 160–1600 CFU 100 ml−1 beer in the end‐point PCR reactions and ≤160 CFU 100 ml−1 beer in the real‐time PCR reactions. More cells (from 16 to 3200) were needed for detection in the presence of brewer's yeast cells. Conclusions: Automated ribotyping is a useful tool to characterize and identify O. proteus and H. alvei isolates. The designed primers are suitable for the rapid detection of O. proteus biotype 1 and H. alvei in brewery samples by PCR. Significance and Impact of the Study: Automated ribotyping and PCR could improve microbiological quality control in breweries by facilitating the detection, identification and tracing of spoilage bacteria.

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“…We have sequenced the 16SrDNA of a biotype II strain of O. proteus (Accession number: AY077753) and designed specific primers for rapid detection and identification of this organism. Biotype I strains of this organism were not amplified by the primer sets; however, this is not Prest et al 1994). The method was highly specific and did not produce amplicons from related species like Ent.…”

Section: Discussionmentioning

confidence: 97%

“…Until now detection and identification of O. proteus isolates has involved extensive culturing and enumeration techniques. Although molecular methods exist in the form of ERIC PCR and plasmid profiling (Prest et al 1994) these are thought to be too complicated to implement in most brewery QC laboratories. PCR analysis is becoming more attractive to QC laboratories and work is currently being conducted in this laboratory and others to help implement this.…”

Section: Discussionmentioning

confidence: 99%

“…Obesumbacterium proteus is an important brewery contaminant that has been isolated from pitching yeast, wort and early fermentation products (Case 1965;Priest and Hough 1974). Obesumbacterium proteus is not normally found in the final product (beer) because it cannot survive in conditions of high ethanol and at pH values below 3AE9 (Prest et al 1994). During the fermentation process O. proteus grows rapidly causing the rate of fermentation to decrease and can lead to an inferior product of high specific gravity and high pH.…”

Section: Introductionmentioning

confidence: 99%

“…Obesumbacterium proteus is an important brewery contaminant that has been isolated from pitching yeast, wort and early fermentation products (Case 1965; Priest and Hough 1974). Obesumbacterium proteus is not normally found in the final product (beer) because it cannot survive in conditions of high ethanol and at pH values below 3·9 (Prest et al . 1994).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Rapid detection of Obesumbacterium proteus from yeast and wort using polymerase chain reaction

Maugueret

¹

,

Walker

²

2002

Lett Appl Microbiol

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Aims: To design and evaluate PCR primers for the rapid detection of Obesumbacterium proteus. Methods and Results: The 16S rDNA from a wild-type Obesumbacterium proteus biotype II isolate was sequenced and the resultant data used to produce specific primers for this organism. These primers discriminated between biotype I (nonbrewery) and biotype II isolates. In addition, the primers were able to detect Obesumbacterium proteus in wort and in yeast slurries in the presence of competitive bacteria. The primers were tested against a range of other beer spoilage bacteria for any cross-reactions. None were detected. Conclusions: Obesumbacterium proteus primers can detect this contaminant without generating cross-reactions to related species. Significance and Impact of the Study: The primers generated in this study can now be used for PCR detection assays that will contribute to early detection of this important process contaminant.

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Obesumbacterium

Farmer¹,

Brenner²

2015

Bergey's Manual of Systematics of Archaea and Bacteria

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O.be' sum.bac.te' ri.um . L. neut. adj. obesum fat; L. neut. n. bacterium rod; M.L. neut. n. Obesumbacterium a fat, rod‐shaped bacterium. Proteobacteria / Gammaproteobacteria / Enterobacteriales / Enterobacteriaceae / Obesumbacterium Pleomorphic rods 0.8–2.0 × 1.5–100 μm (short, “fat” rods predominate when grown in beer wort with live yeasts; long pleomorphic rods usually predominate when grown in most bacteriological media; some strains have been reported to display a branching cell morphology), conforming to the general definition of the family Enterobacteriaceae . Gram negative. Nonmotile . Facultatively anaerobic. Slow growing, forming colonies <0.5 mm in diameter on ordinary plating media at 24 h. Optimal growth temperature is 25–32°C; growth at 37°C is comparatively poor. Acid formed from D ‐glucose and D ‐mannose; very few other carbohydrates are fermented . Gas formation during fermentation appears to be variable (original description says gas is produced, but none of the strains studied produced gas). Lysine decarboxylase is positive . Nitrate is reduced to nitrite. Many biochemical tests normally used for differentiation of Enterobacteriaceae are negative or delayed positive (3–7 d at 36°C). One of only three genera (along with Hafnia and Pragia ) in Enterobacteriaceae to contain only gluconate dehydrogenase in its D ‐glucose oxidation pathway (Bouvet et al., 1989). Occurs as a brewery contaminant that can survive and grow in the presence of live yeasts during beer production. Not isolated from human clinical specimens; no evidence of pathogenicity for humans or animals. The genus has a single species, O . proteus , with two defined biogroups (1 and 2). The description of the genus above is a “composite description” based on data for both biogroups. However, the two biogroups are actually distinct species that are phenotypically different and only distantly related by DNA–DNA hybridization (Brenner, 1981). O . proteus biogroup 1 is actually a biogroup of Hafnia alvei (Brenner, 1981), and will be referred to as H . alvei biogroup 1 (Farmer et al., 1985a) in this chapter. The mol % G + C of the DNA is : 48–49. Type species : Obesumbacterium proteus (Shimwell and Grimes 1936) Shimwell 1963, 759 ( Flavobacterium proteum (sic) Shimwell and Grimes 1936, 348.)

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Biochemical and Molecular Characterization of Obesumbacterium proteus , a Common Contaminant of Brewing Yeasts

Cited by 19 publications

References 24 publications

Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1

Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1

Rapid detection of Obesumbacterium proteus from yeast and wort using polymerase chain reaction

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