Pediococcus parvulus gtf Gene Encoding the GTF Glycosyltransferase and Its Application for Specific PCR Detection of β-d-Glucan–Producing Bacteria in Foods and Beverages

2006 ◽  
Vol 69 (1) ◽  
pp. 161-169 ◽  
Author(s):  
MARIA LAURA WERNING ◽  
IDOIA IBARBURU ◽  
MARIA TERESA DUEÑAS ◽  
ANA IRASTORZA ◽  
JESÚS NAVAS ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Blot Hybridization ◽  
Pcr Amplification ◽  
Southern Blot Hybridization ◽  
Oenococcus Oeni ◽  
Alcoholic Beverages ◽  
Gene Encoding ◽  
Specific Pcr ◽  
Genomic Locations

Exopolysaccharide production by lactic acid bacteria is beneficial in the dairy and oat-based food industries and is used to improve the texture of the fermented products. However, β-D-glucan–producing bacteria are considered spoilage microorganisms in alcoholic beverages because their secreted exopolysaccharides alter the viscosity of cider, wine, and beer, rendering them unpalatable. The plasmidic glycosyltransferase (gtf) gene of the Pediococcus parvulus 2.6 strain isolated from ropy cider has been cloned and sequenced, and its GTF product was functionally expressed in Streptococcus pneumoniae. The GTF protein, which has glycosyltransferase activity, belongs to the COG1215 membrane-bound glycosyltransferase family, and agglutination tests revealed that the enzyme enables S. pneumoniae to synthesize β-D-glucan. PCR amplification and Southern blot hybridization showed that the gtf gene is also present at different genomic locations in the β-d-glucan producers Lacto-bacillus diolivorans G77 and Oenococcus oeni I4 strains, also isolated from ropy cider. A PCR assay has been developed for the detection of exopolysaccharide-producing bacteria. Forward and reverse primers, included respectively in the coding sequences of the putative glycosyltransferase domain and the fifth trans-membrane segment of the GTF, were designed. Analysis of 76 ropy and nonropy lactic acid bacteria validated the method for specific detection of β-d-glucan homopolysaccharide producer Pediococcus, Lactobacillus, and Oenococcus strains. The limit of the assay in cider was 3 × 102 CFU/ml. This molecular method can be useful for the detection of ropy bacteria in cider before spoilage occurs, as well as for isolation of new exopolysaccharide-producing strains of industrial interest.

scholarly journals Ethylphenol Formation byLactobacillus plantarum: Identification of the Enzyme Involved in the Reduction of Vinylphenols

2018 ◽  
Vol 84 (17) ◽  
Author(s):  
Laura Santamaría ◽  
Inés Reverón ◽  
Félix López de Felipe ◽  
Blanca de las Rivas ◽  
Rosario Muñoz
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bacterial Species ◽  
Alcoholic Beverages ◽  
Genetic Identification ◽  
Detection Methods ◽  
Fermented Foods ◽  
Content Type ◽  
Volatile Phenol ◽  
Off Flavors

ABSTRACTEthylphenols are strong odorants produced by microbial activity that are described as off flavors in several foods.Lactobacillus plantarumis a lactic acid bacterial species able to produce ethylphenols by the reduction of vinylphenols during the metabolism of hydroxycinnamic acids. However, the reductase involved has not been yet uncovered. In this study, the involvement in vinylphenol reduction of a gene encoding a putative reductase (lp_3125) was confirmed by the absence of reduction activity in the Δlp_3125knockout mutant. The protein encoded bylp_3125, VprA, was recombinantly produced inEscherichia coli. VprA was assayed against vinylphenols (4-vinylphenol, 4-vinylcatechol, and 4-vinylguaiacol), and all were reduced to their corresponding ethylphenols (4-ethylphenol, 4-ethylcatechol, and 4-ethylguaiacol). PCR and high-performance liquid chromatography (HPLC) detection methods revealed that the VprA reductase is not widely distributed among the lactic acid bacteria studied and that only the bacteria possessing thevprAgene were able to produce ethylphenol from vinylphenol. However, all the species belonging to theL. plantarumgroup were ethylphenol producers. The identification of theL. plantarumVprA protein involved in hydroxycinnamate degradation completes the route of degradation of these compounds in lactic acid bacteria.IMPORTANCEThe presence of volatile phenols is considered a major organoleptic defect of several fermented alcoholic beverages. The biosynthesis of these compounds has been mainly associated withBrettanomyces/Dekkerayeasts. However, the potential importance of lactic acid bacteria in volatile phenol spoilage is emphasized by reports describing a faster ethylphenol production by these bacteria than by yeasts. The genetic identification of the bacterial vinylphenol reductase involved in volatile phenol production provides new insights into the role of lactic acid bacteria in the production of these off flavors. The development of a molecular method for the detection of ethylphenol-producing bacteria could be helpful to design strategies to reduce the bacterial production of vinylphenols in fermented foods.

scholarly journals Isolation, identification and antibiotic susceptibility of nis+ Lactococcus lactis from dairy and non-dairy sources

2013 ◽  
Vol 31 (No. 4) ◽  
pp. 323-331 ◽  
Author(s):  
P. Khemariya ◽  
S. Singh ◽  
G. Nath ◽  
A.K. Gulati
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Antimicrobial Agent ◽  
Lactococcus Lactis ◽  
Antibiotic Susceptibility ◽  
Pcr Amplification ◽  
Food Borne ◽  
Antibiotic Susceptibility Test ◽  
Different Types ◽  
Zones Of Inhibition

Eight isolates of Lactococcus lactis subsp. lactis were isolated and identified by phenotypic and molecular characterisation out of 23 isolates of lactic acid bacteria (LAB) from different dairy and non-dairy sources. Out of eight strains, four were obtained from dairy and four from non-dairy sources. All eight strains of L. lactis subsp. lactis were able to produce zones of inhibition against the Lactobacillus acidophilus NCDC 015. The antimicrobial agent produced by the isolates inhibited the growth of a range of related lactic acid bacteria and certain Gram positive food-borne microorganisms. The antimicrobial agent, i.e. nisin, produced by the strains was confirmed by PCR amplification of nisin gene sequences of 174 bp size. Antibiotic susceptibility test to 21 different types of antibiotics was evaluated. All the isolates were resistant to fosfomycin, cefepime, amikacin, kanamycin, neomycin, nalidixic acid, pipemidic acid, norfloxacin, sulphadiazine, colistin, polymixin, teicoplanin, nystatin, and amphotericin B but susceptible to ampicillin, erythromycin, spiramycin, spectinomycin, ciprofloxacin, rifampicin, and trimethoprim.  

scholarly journals Differential Real-Time PCR Assay for Enumeration of Lactic Acid Bacteria in Wine

2005 ◽  
Vol 71 (12) ◽  
pp. 8954-8957 ◽  
Author(s):  
Ezekiel T. Neeley ◽  
Trevor G. Phister ◽  
David A. Mills
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Real Time ◽  
Real Time Pcr ◽  
Malolactic Fermentation ◽  
Oenococcus Oeni ◽  
Pcr Assay ◽  
Wine Production ◽  
Bacterial Populations ◽  
Wine Spoilage

ABSTRACT Oenococcus oeni is often employed to perform the malolactic fermentation in wine production, while nonoenococcal lactic acid bacteria often contribute to wine spoilage. Two real-time PCR assays were developed to enumerate the total, and nonoenococcal, lactic acid bacterial populations in wine. Used together, these assays can assess the spoilage risk of juice or wine from lactic acid bacteria.

scholarly journals Use of the alr Gene as a Food-Grade Selection Marker in Lactic Acid Bacteria

2002 ◽  
Vol 68 (11) ◽  
pp. 5663-5670 ◽  
Author(s):  
Peter A. Bron ◽  
Marcos G. Benchimol ◽  
Jolanda Lambert ◽  
Emmanuelle Palumbo ◽  
Marie Deghorain ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Competitive Inhibitor ◽  
Selection Marker ◽  
Wild Type ◽  
Gene Encoding ◽  
Food Grade ◽  
Selection For ◽  
Isogenic Mutants ◽  
Type Strains

ABSTRACT Both Lactococcus lactis and Lactobacillus plantarum contain a single alr gene, encoding an alanine racemase (EC 5.1.1.1), which catalyzes the interconversion of d-alanine and l-alanine. The alr genes of these lactic acid bacteria were investigated for their application as food-grade selection markers in a heterologous complementation approach. Since isogenic mutants of both species carrying an alr deletion (Δalr) showed auxotrophy for d-alanine, plasmids carrying a heterologous alr were constructed and could be selected, since they complemented d-alanine auxotrophy in the L. plantarum Δalr and L. lactis Δalr strains. Selection was found to be highly stringent, and plasmids were stably maintained over 200 generations of culturing. Moreover, the plasmids carrying the heterologous alr genes could be stably maintained in wild-type strains of L. plantarum and L. lactis by selection for resistance to d-cycloserine, a competitive inhibitor of Alr (600 and 200 μg/ml, respectively). In addition, a plasmid carrying the L. plantarum alr gene under control of the regulated nisA promoter was constructed to demonstrate that d-cycloserine resistance of L. lactis is linearly correlated to the alr expression level. Finally, the L. lactis alr gene controlled by the nisA promoter, together with the nisin-regulatory genes nisRK, were integrated into the chromosome of L. plantarum Δalr. The resulting strain could grow in the absence of d-alanine only when expression of the alr gene was induced with nisin.

scholarly journals Shifts in Abundance and Diversity of Mobile Genetic Elements after the Introduction of Diverse Pesticides into an On-Farm Biopurification System over the Course of a Year

2014 ◽  
Vol 80 (13) ◽  
pp. 4012-4020 ◽  
Author(s):  
Simone Dealtry ◽  
Peter N. Holmsgaard ◽  
Vincent Dunon ◽  
Sven Jechalke ◽  
Guo-Chun Ding ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Pcr Amplification ◽  
Southern Blot Hybridization ◽  
Mobile Genetic Elements ◽  
Replication Initiation ◽  
Genetic Elements ◽  
Class 1 ◽  
Genes Encoding ◽  
Abundance And Diversity ◽  
And Shifts

ABSTRACTBiopurification systems (BPS) are used on farms to control pollution by treating pesticide-contaminated water. It is assumed that mobile genetic elements (MGEs) carrying genes coding for enzymes involved in degradation might contribute to the degradation of pesticides. Therefore, the composition and shifts of MGEs, in particular, of IncP-1 plasmids carried by BPS bacterial communities exposed to various pesticides, were monitored over the course of an agricultural season. PCR amplification of total community DNA using primers targeting genes specific to different plasmid groups combined with Southern blot hybridization indicated a high abundance of plasmids belonging to IncP-1, IncP-7, IncP-9, IncQ, and IncW, while IncU and IncN plasmids were less abundant or not detected. Furthermore, the integrase genes of class 1 and 2 integrons (intI1,intI2) and genes encoding resistance to sulfonamides (sul1,sul2) and streptomycin (aadA) were detected and seasonality was revealed. Amplicon pyrosequencing of the IncP-1trfAgene coding for the replication initiation protein revealed high IncP-1 plasmid diversity and an increase in the abundance of IncP-1β and a decrease in the abundance of IncP-1ε over time. The data of the chemical analysis showed increasing concentrations of various pesticides over the course of the agricultural season. As an increase in the relative abundances of bacteria carrying IncP-1β plasmids also occurred, this might point to a role of these plasmids in the degradation of many different pesticides.

scholarly journals Isolation and Identification of Microorganisms Including Lactic Acid Bacteria and Their Use in Microbial Deacidification of Wines from Domestic Vineyards

2013 ◽  
Vol 62 (3) ◽  
pp. 331-334
Author(s):  
IWONA DROŻDŻ ◽  
MAŁGORZATA MAKAREWICZ ◽  
TADEUSZ TUSZYŃSKI
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Acidophilus ◽  
Oenococcus Oeni ◽  
Lactobacillus Delbrueckii ◽  
The Other ◽  
Total Acidity ◽  
Isolation And Identification ◽  
Volatile Acidity ◽  
Other Hand

The aim of this study was to identify various bacteria isolated from grapes and their wines. Additionally we investigated the capacity of lactic acid bacteria for microbiological deacidification of wines produced in Poland. We have identified Oenococcus oeni, Lactobacillus acidophilus and Lactobacillus delbrueckii. During the microbial deacidification process, we observed decreases of total acidity and increases of volatile acidity, with statistically significant changes noted for O. oeni in Marechal Foch and Seyval Blanc, and for Lb. acidophilus in Frontenac. On the other hand, a statistically significant increase in pH was observed in Marechal Foch and Seyval Blanc following deacidification by O. oeni.

scholarly journals RELEVANCE OF ETHYL LACTATE AMONG THE CACHAÇA ESTERS

2021 ◽  
Vol 2 (6) ◽  
pp. e26467
Author(s):  
Amazile Biagionei Maia ◽  
Lorena Simão Marinho ◽  
David Lee Nelson
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Ethyl Acetate ◽  
Sensory Quality ◽  
Chemical Characterization ◽  
Ethyl Lactate ◽  
Alcoholic Beverages ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cane Juice

ABSTRACT The advancement in the technology of cachaça ─ sugarcane spirit produced in the Brazilian territory ─ has led to the search for parameters that allow the improvement of its chemical characterization and sensory specificities. Esters are important aroma agents, but their routine monitoring, as endorsed in Brazilian legislation, is restricted to the quantification of ethyl acetate, which is formed through the metabolism of yeast (Saccharomyces cerevisiae). However, being produced in a rural environment using fresh cane juice, fermentation of “cachaça de alembic” involves the presence and activity of lactic acid-producing bacteria, making the presence of ethyl lactate among the cachaça esters predictable. In this work, the levels of ethyl lactate and ethyl acetate were compared in 247 samples, corresponding to 56 brands produced in eleven Brazilian states. Ethyl lactate was found in levels that significantly alter the result of the quantitative participation of esters in the composition of cachaça. This fact is especially relevant considering that lactic acid bacteria are GRAS and widely recognized as a resource for improving the sensory quality of wines and other alcoholic beverages. Thus, studies on the contribution of lactic acid bacteria to the sensory quality of cachaça should be encouraged. Ethyl lactate, in addition to signaling a chemical specificity naturally occurring in cachaça, is a marker of other possible chemical and sensory peculiarities whose research should be stimulated.

Primers and a Specific DNA Probe for Detecting Lactic Acid Bacteria Producing 3-Hydroxypropionaldehyde from Glycerol in Spoiled Ciders

2001 ◽  
Vol 64 (6) ◽  
pp. 833-837 ◽  
Author(s):  
OLIVIER CLAISSE ◽  
ALINE LONVAUD-FUNEL
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Polymerase Chain Reaction Amplification ◽  
Klebsiella Oxytoca ◽  
Blot Hybridization ◽  
Dna Probe ◽  
Dot Blot ◽  
Glycerol Dehydratase ◽  
Polymerase Chain ◽  
Dna Primers

Of the 40 strains isolated from several spoiled ciders where glycerol was degraded, 36 were identified as Lactobacillus collinoides, three were Lactobacillus hilgardii, and one was Lactobacillus mali. However, only 30 L. collinoides and two L. hilgardii could degrade glycerol. The glycerol dehydratase activity was shown. The main product of the transformation was 1,3 propanediol. Two DNA primers GD1 and GD2 were chosen in the region encoding one of the subunits of glycerol dehydratase of Citrobacter freundii, Klebsiella pneumoniae, Klebsiella oxytoca, Salmonella Typhimurium, and Clostridium pasteurianum. A 279-bp amplicon in polymerase chain reaction amplification was obtained with the genomic L. collinoides IOEB 9527 DNA as template. The amino acid sequence deduced from the amplicon DNA sequence showed a very high similarity and identity with the gene of gram-negative and C. pasteurianum species. After labeling, the amplicon was used as DNA probe in dot-blot hybridization with the genomic DNA of all the tested strains. Only strains that could degrade glycerol hybridized. Moreover, polymerase chain reactions using GD1 and GD2 revealed only glycerol dehydratase genes of positive L. collinoides and L. hilgardii strains. The primers and the amplicon proved to be suitable and reliable tools to detect the lactic acid bacteria involved in the deterioration of cider.

Antioxidant properties of wine lactic acid bacteria: Oenococcus oeni

2015 ◽  
Vol 99 (12) ◽  
pp. 5189-5202 ◽  
Author(s):  
Jing Su ◽  
Tao Wang ◽  
Ying-Ying Li ◽  
Jing Li ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Antioxidant Properties ◽  
Oenococcus Oeni

scholarly journals Oenococcus kitaharae sp. nov., a non-acidophilic and non-malolactic-fermenting oenococcus isolated from a composting distilled shochu residue

2006 ◽  
Vol 56 (10) ◽  
pp. 2345-2348 ◽  
Author(s):  
Akihito Endo ◽  
Sanae Okada
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Novel Species ◽  
Malolactic Fermentation ◽  
Oenococcus Oeni ◽  
Rrna Gene ◽  
16S Rrna Gene Sequences ◽  
Phenotypic Characteristics ◽  
Optimum Ph ◽  
The 16S Rrna Gene

Six strains of lactic acid bacteria were isolated in Japan from a composting distilled shochu residue. The six isolates grew poorly on MRS agar and slowly in MRS broth. The 16S rRNA gene sequences did not show high levels of similarity to those of the recognized species of lactic acid bacteria, and formed a subcluster within the cluster comprising obligately heterofermentative lactic acid bacteria closely related to Oenococcus oeni. The levels of DNA–DNA relatedness revealed that the isolates belonged to the same taxon and were genetically separate from O. oeni. Furthermore, various phenotypic characteristics such as the optimum pH for growth, malolactic fermentation and resistance to 10 % ethanol revealed that the isolates are distinguishable from O. oeni. On the basis of their phylogenetic and phenotypic characteristics, the isolates represent a novel species, for which the name Oenococcus kitaharae sp. nov. is proposed. The type strain is NRIC 0645T (=JCM 13282T=DSM 17330T).

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