Detection of beer spoilage bacteria Pectinatus and Megasphaera with acridinium ester labelled DNA probes using a hybridisation protection assay

2014 ◽  
Vol 96 ◽  
pp. 25-34 ◽  
Author(s):  
A.D. Paradh ◽  
A.E. Hill ◽  
W.J. Mitchell
Keyword(s):  
Dna Probes ◽  
Protection Assay ◽  
Spoilage Bacteria ◽  
Beer Spoilage ◽  
Acridinium Ester

scholarly journals Development of a Hypersensitive Detection Method for Human Parvovirus B19 DNA

2000 ◽  
Vol 38 (3) ◽  
pp. 1241-1243 ◽  
Author(s):  
Kazuaki Sato ◽  
Eiji Matsuda ◽  
Keiichi Kamisango ◽  
Hiroaki Iwasaki ◽  
Shuzo Matsubara ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Detection Method ◽  
Parvovirus B19 ◽  
Dna Probes ◽  
Human Parvovirus B19 ◽  
Protection Assay ◽  
Acridinium Ester

A new detection method for human parvovirus B19 DNA was established using PCR coupled with a hybridization protection assay. The amplified product was detected using acridinium ester-labeled DNA probes. By this method, a few copies of B19 DNA were detected in human serum albumin.

Identification of novel horA-harbouring bacteria capable of spoiling beer

2008 ◽  
Vol 54 (4) ◽  
pp. 321-325 ◽  
Author(s):  
Monique Haakensen ◽  
Barry Ziola
Keyword(s):  
Drug Resistance ◽  
Bacillus Cereus ◽  
Bacillus Licheniformis ◽  
Staphylococcus Epidermidis ◽  
Multi Drug Resistance ◽  
Bacterial Isolates ◽  
First Finding ◽  
Spoilage Bacteria ◽  
Beer Spoilage ◽  
Respective Species

An ATP-binding cassette (ABC) multi-drug resistance (MDR) gene was found in 4 Gram-positive bacterial isolates of environmental origin and found capable of spoiling beer. The bacteria isolated were Bacillus cereus , Bacillus licheniformis , Paenibacillus humicus , and Staphylococcus epidermidis ; all of which were previously unappreciated as beer-spoilage bacteria. The MDR gene found in these bacteria has less than 37% similarity to known ABC MDR proteins described for Bacillus and Staphylococcus , and this is the first finding of an ABC MDR gene in the genus Paenibacillus . The sequenced region of the gene was translated and compared phylogenetically with the closest GenBank matches of the respective species and the closest GenBank matches overall. The ABC MDR proteins from these isolates were found to cluster among known sequences of HorA, sharing 99.5% identity within the sequenced region. In the beer-spoilage-associated genera Lactobacillus and Pediococcus , the presence of the MDR gene horA correlates with the ability to grow in beer. As the unique horA-harbouring isolates described here are capable of growing in beer, it is likely that the presence of the horA gene likewise confers hop resistance to these organisms.

BEER-SPOILAGE BACTERIA AND THEIR CONTROL WITH A PHOSPHORIC ACID – AMMONIUM PERSULFATE WASH

1965 ◽  
Vol 11 (2) ◽  
pp. 309-318 ◽  
Author(s):  
Stanley Bah ◽  
W. E. McKeen
Keyword(s):  
Phosphoric Acid ◽  
Treatment Time ◽  
Ammonium Persulfate ◽  
Acetobacter Aceti ◽  
Lager Yeast ◽  
Great Saving ◽  
Spoilage Bacteria ◽  
Plant Equipment ◽  
Beer Spoilage ◽  
The One

Flavobacterium proteus, Acetobacter aceti, Acetobacter capsulatum, Pediococcus cerevisiae, and Lactobacillus pastorianus were isolated from contaminated yeast in certain Canadian breweries. F. proteus was most prevalent in ale yeast, while L. pastorianus and P. cerevisiae were most prevalent in lager yeast but the plant also affected the prevalence of the species.A phosphoric acid – ammonium persulfate wash was developed which has proved very effective in the breweries in which it has been used since 1960. Treatment time is very short (2 hours) and consequently a great saving in plant equipment and labor has resulted.The fermentative ability and viability of Saccharomyces cerevisiae is not affected nor is it killed even when it is treated with a phosphoric acid – ammonium persulfate wash that is six times as strong as the one recommended for use in the brewery.

scholarly journals Staphylococcus aureus DNA ligase: characterization of its kinetics of catalysis and development of a high-throughput screening compatible chemiluminescent hybridization protection assay

2004 ◽  
Vol 383 (3) ◽  
pp. 551-559 ◽  
Author(s):  
Sheraz GUL ◽  
Richard BROWN ◽  
Earl MAY ◽  
Marie MAZZULLA ◽  
Martin G. SMYTH ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
High Throughput ◽  
High Throughput Screening ◽  
Dna Ligase ◽  
Protection Assay ◽  
Dna Ligases ◽  
Enzyme Substrate ◽  
Ic50 Values ◽  
Acridinium Ester

DNA ligases are key enzymes involved in the repair and replication of DNA. Prokaryotic DNA ligases uniquely use NAD+ as the adenylate donor during catalysis, whereas eukaryotic enzymes use ATP. This difference in substrate specificity makes the bacterial enzymes potential targets for therapeutic intervention. We have developed a homogeneous chemiluminescence-based hybridization protection assay for Staphylococcus aureus DNA ligase that uses novel acridinium ester technology and demonstrate that it is an alternative to the commonly used radiometric assays for ligases. The assay has been used to determine a number of kinetic constants for S. aureus DNA ligase catalysis. These included the Km values for NAD+ (2.75±0.1 μM) and the acridinium-ester-labelled DNA substrate (2.5±0.2 nM). A study of the pH-dependencies of kcat, Km and kcat/Km has revealed values of kinetically influential ionizations within the enzyme–substrate complexes (kcat) and free enzyme (kcat/Km). In each case, the curves were shown to be composed of one kinetically influential ionization, for kcat, pKa=6.6±0.1 and kcat/Km, pKa=7.1±0.1. Inhibition characteristics of the enzyme against two Escherichia coli DNA ligase inhibitors have also been determined with IC50 values for these being 3.30±0.86 μM for doxorubicin and 1.40±0.07 μM for chloroquine diphosphate. The assay has also been successfully miniaturized to a sufficiently low volume to allow it to be utilized in a high-throughput screen (384-well format; 20 μl reaction volume), enabling the assay to be used in screening campaigns against libraries of compounds to discover leads for further drug development.

Serogroups of the beer spoilage bacterium Megasphaera cerevisiae correlate with the molecular weight of the major EDTA-extractable surface protein

2000 ◽  
Vol 46 (2) ◽  
pp. 95-100 ◽  
Author(s):  
Barry Ziola ◽  
Lori Gee ◽  
Nancy N Berg ◽  
Sun Y Lee
Keyword(s):  
Ethylenediaminetetraacetic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Surface Protein ◽  
Cross Reactivity ◽  
Surface Reactivity ◽  
Surface Proteins ◽  
Bacterial Surface ◽  
Spoilage Bacteria ◽  
Beer Spoilage

Megasphaera cerevisiae is a Gram-negative obligate anaerobe that causes turbidity and off-flavour and aroma in beer. Seven isolates of M. cerevisiae were obtained worldwide, and their extractable surface antigens were focused upon to determine if there is more than one serogroup of this bacterium. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of ethylenediaminetetraacetic acid (EDTA) bacterial extracts revealed a predominant protein with apparent molecular weights of 46 000, 45 000, and 43 000 for three, two, and two isolates, respectively. When mouse antiserum generated against any of the EDTA extracts was reacted with denatured bacterial proteins in immunoblots, all bacterial isolates exhibited extensive cross-reactivity involving three antigens, one being the major EDTA-extractable protein. In contrast, when the sera were tested for surface reactivity with intact bacteria, three cross-reactivity groups were observed, with the groups individually comprised of bacteria having the same size major EDTA-extractable surface protein. When BALB/c mice immunized with a bacterium from each of the three serogroups were used for monoclonal antibody (Mab) hybridoma production, bacterial surface-reactive Mabs were obtained whose reactivities parallel the three polyclonal antibody-defined serogroups. Through combining these surface-reactive Mabs, it will be possible to rapidly detect and identify beer contamination by M. cerevisiae belonging to any serogroup. Key words: beer spoilage bacteria, Megasphaera cerevisiae, monoclonal antibodies, surface proteins, serogroups.

Isolation of a Virulent Lactobacillus brevis Phage and Its Application in the Control of Beer Spoilage

2011 ◽  
Vol 74 (12) ◽  
pp. 2157-2161 ◽  
Author(s):  
THERESE DEASY ◽  
JENNIFER MAHONY ◽  
HORST NEVE ◽  
KNUT J. HELLER ◽  
DOUWE VAN SINDEREN
Keyword(s):  
Electron Microscopy ◽  
Lactobacillus Plantarum ◽  
Bacterial Contamination ◽  
Lactobacillus Brevis ◽  
Virulent Phage ◽  
Spoilage Bacteria ◽  
Food And Beverage ◽  
Beer Spoilage ◽  
Bacterial Spoilage

Beer quality can be compromised by the growth of certain lactobacilli, in particular Lactobacillus brevis and Lactobacillus plantarum, and various strategies have been used to control such bacterial spoilage. Biocontrol by means of bacteriophage is a reemerging approach for the suppression of spoilage bacteria in food and beverage matrices. A virulent phage capable of infecting L. brevis beer-spoilage strains was isolated and morphologically assessed by electron microscopy. The myophage SA-C12 was shown to be stable in beer and capable of controlling the growth of its host, L. brevis strain 56, in commercial beer. The results of this study indicate that bacteriophage-based treatments may be used as an alternative and natural strategy for the control of bacterial contamination of beer.

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