Comparative analysis of conserved genetic markers and adjacent DNA regions identified in beer-spoilage lactic acid bacteria

Suzuki, Koji; Ozaki, Kazutaka; Yamashita, Hiroshi

doi:10.1111/j.1472-765x.2004.01572.x

Cited by 31 publications

(29 citation statements)

References 20 publications

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“…99% nucleotide sequence identities were found in these approximately 8.2-kb DNA regions (30). Therefore we assumed that the ORF 1 to 7 region transfers between strains as a composite transposon with ORF 3 and ORF 7, encoding putative transposase and nicking enzyme, being the mediators of horizontal gene transfer.…”

Section: Discussionmentioning

confidence: 99%

“…This possibility led us to reexamine the ORF 1 to 7 region that was implicated in the hop resistance ability of LAB and considered to carry a gene that encodes a proton motive forcedependent multidrug transporter (30,33,35). Southern blot analysis indicates that L. lindneri DSM 20692 possesses homologs of ORF 1 and ORF 2, renamed horB and horC, respectively, on a plasmid and this DNA region was found to be lost, concomitant with the loss of hop resistance ability of DSM 20692 (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The partial loss of pRH45II, coupled with the loss of pRH45, resulted in complete loss of the beer spoilage ability of L. brevis ABBC45 (33). The excised DNA region of pRH45II contained the ORF 1 to 7 region that was implicated in horA-independent hop resistance mechanism (30,33). Biochemical data indicate that a proton motive forcedependent multidrug transporter mediates horA-independent hop resistance (35).…”

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confidence: 99%

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Isolation of a Hop-Sensitive Variant of Lactobacillus lindneri and Identification of Genetic Markers for Beer Spoilage Ability of Lactic Acid Bacteria

Suzuki

Iijima

Ozaki

et al. 2005

Appl Environ Microbiol

111

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We have isolated a hop-sensitive variant of the beer spoilage bacterium Lactobacillus lindneri DSM 20692. The variant lost a plasmid carrying two contiguous open reading frames (ORF s) designated horB L and horC L that encode a putative regulator and multidrug transporter presumably belonging to the resistance-nodulation-cell division superfamily. The loss of hop resistance ability occurred with the loss of resistance to other drugs, including ethidium bromide, novobiocin, and cetyltrimethylammonium bromide. PCR and Southern blot analysis using 51 beer spoilage strains of various species of lactic acid bacteria (LAB) revealed that 49 strains possessed homologs of horB and horC. No false-positive results have been observed for nonspoilage LAB or frequently encountered brewery isolates. These features are superior to those of horA and ORF 5, previously reported genetic markers for determining the beer spoilage ability of LAB. It was further shown that the combined use of horB/horC and horA is able to detect all 51 beer spoilage strains examined in this study. Furthermore sequence comparison of horB and horC homologs identified in four different beer spoilage species indicates these homologs are 96.6 to 99.5% identical, which is not typical of distinct species. The wide and exclusive distribution of horB and horC homologs among beer spoilage LAB and their sequence identities suggest that the hop resistance ability of beer spoilage LAB has been acquired through horizontal gene transfer. These insights provide a foundation for applying trans-species genetic markers to differentiating beer spoilage LAB including previously unencountered species.Beer has been recognized as a beverage with high microbiological stability. Among the beer spoilers, several species of lactic acid bacteria (LAB) are reported to be responsible for approximately 70% of spoilage incidents caused by microorganisms (2, 3). For this reason, species-specific identification methods based on PCR have been widely evaluated for potential applications to microbiological quality control (6,19,41,42,44). Although species-specific PCR tests are rapid and reasonably accurate, there are two problems for applying this approach to the quality control of breweries.One problem is that the species-specific method is unable to distinguish intraspecies differences between beer spoilage strains and nonspoilage strains (9,26,28,35). Hop compounds added to confer bitter flavor are reported to exert an antibacterial effect by acting as proton ionophores and dissipate transmembrane pH gradient, which prevents gram-positive bacteria, including most LAB, from growing in beer (24,25,27,28,40). Hop resistance ability has been known as the distinguishing character of beer spoilage strains of LAB and nonspoilage strains typically exhibit hop resistance ability considerably weaker than that of beer spoilage strains belonging to the same species (1,9,28,34,35). The presence of nonspoilage strains within a beer spoilage species inevitably leads to false-positive results as long ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Isolation of a Hop-Sensitive Variant of Lactobacillus lindneri and Identification of Genetic Markers for Beer Spoilage Ability of Lactic Acid Bacteria

Suzuki

Iijima

Ozaki

et al. 2005

Appl Environ Microbiol

111

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“…A key factor in hop resistance is the plasmid-encoded, ATP-dependent transporter protein HorA, which purges hop compounds from the cell (223). Another plasmid-encoded multidrug transporter, ORF5, has been shown to confer hop resistance across multiple species of LAB (209). In addition, resistant cells upregulate expression of the hop-inducible cation transporter HitA, which may facilitate manganese transport into hop-stressed cells despite proton gradient dissipation (224).…”

Section: Pediococcus Damnosus Pediococcus Inopinatus Pediococcus Dementioning

confidence: 99%

The Microbiology of Malting and Brewing

Bokulich

Bamforth

2013

Microbiol Mol Biol Rev

267

206

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SUMMARY Brewing beer involves microbial activity at every stage, from raw material production and malting to stability in the package. Most of these activities are desirable, as beer is the result of a traditional food fermentation, but others represent threats to the quality of the final product and must be controlled actively through careful management, the daily task of maltsters and brewers globally. This review collates current knowledge relevant to the biology of brewing yeast, fermentation management, and the microbial ecology of beer and brewing.

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“…Therefore, the potential applications of these genetic markers have been suggested to the comprehensive species-independent identification of beer-spoilage LAB strains. The horizontal transfer hypothesis of hop-resistance genes has been based on the sequence identities of the horA and horC-carrying DNA regions found in L. brevis, L. lindneri, L. paracollinoides and P. damnosus 17,[22][23][24] . Therefore we attempted to characterize the horA and horC-containing DNA regions in P. inopinatus and L. backi.…”

Section: Analysis Of the Hora-containing Regions And The Horc-containmentioning

confidence: 99%

Isolation and Identification of Potential Beer-Spoilage Pediococcus inopinatus and Beer-Spoilage Lactobacillus backi Strains Carrying the horA and horC Gene Clusters

Iijima

Suzuki

Asano

et al. 2007

Journal of the Institute of Brewing

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Four beer-spoilage strains, LA20, LA21, LA22 and LA23, were isolated from brewery environments. Based on the 16S rRNA gene sequence, LA20 was identified as Pediococcus inopinatus and the remaining three were identified as Lactobacillus backi. The homologs of horA and horC, the hop resistance genes originally identified in L. brevis ABBC45, were detected simultaneously in LA22 and LA23, while only a horA homolog and a horC homolog were found in LA20 and LA21 respectively. The 5.6 kb DNA regions containing the horA homolog in LA20 and LA22 were almost 99% identical with the corresponding region of ABBC45. Similarly the 8.2 kb regions containing the horC homolog in LA21 and LA22 were more than 99% identical with that of ABBC45. Interestingly the horA-containing 5.6 kb regions in LA20 and LA22 were found to be completely identical despite the distinct genus status. Coupled with the fact that LA20 and LA22 were isolated from the same sampling site, these results, taken collectively, reinforce our hypothesis that horA and horC genes were acquired by beer-spoilage species through horizontal gene transfer and confirm the usefulness of horA and horC as genetic markers for the species-independent determination of beer-spoilage ability in lactic acid bacteria.

show abstract

Comparative analysis of conserved genetic markers and adjacent DNA regions identified in beer-spoilage lactic acid bacteria

Cited by 31 publications

References 20 publications

Isolation of a Hop-Sensitive Variant of Lactobacillus lindneri and Identification of Genetic Markers for Beer Spoilage Ability of Lactic Acid Bacteria

Isolation of a Hop-Sensitive Variant of Lactobacillus lindneri and Identification of Genetic Markers for Beer Spoilage Ability of Lactic Acid Bacteria

The Microbiology of Malting and Brewing

Isolation and Identification of Potential Beer-Spoilage Pediococcus inopinatus and Beer-Spoilage Lactobacillus backi Strains Carrying the horA and horC Gene Clusters

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