scholarly journals Cloning and Characterization of an Intracellular Esterase from the Wine-Associated Lactic Acid Bacterium Oenococcus oeni

2009 ◽  
Vol 75 (21) ◽  
pp. 6729-6735 ◽  
Author(s):  
Krista M. Sumby ◽  
Angela H. Matthews ◽  
Paul R. Grbin ◽  
Vladimir Jiranek
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Ethanol Concentration ◽  
Expression System ◽  
Pcr Primers ◽  
Oenococcus Oeni ◽  
Optimum Temperature ◽  
Reading Frame ◽  
Marked Activity

ABSTRACT We report the cloning and characterization of EstB28, the first esterase to be so characterized from the wine-associated lactic acid bacterium, Oenococcus oeni. The published sequence for O. oeni strain PSU-1 was used to identify putative esterase genes and design PCR primers in order to amplify the corresponding region from strain Ooeni28, an isolate intended for inoculation of wines. In this way a 912-bp open reading frame (ORF) encoding a putative esterase of 34.5 kDa was obtained. The amino acid sequence indicated that EstB28 is a member of family IV of lipolytic enzymes and contains the GDSAG motif common to other lactic acid bacteria. This ORF was cloned into Escherichia coli using an appropriate expression system, and the recombinant esterase was purified. Characterization of EstB28 revealed that the optimum temperature, pH, and ethanol concentration were 40°C, pH 5.0, and 28% (vol/vol), respectively. EstB28 also retained marked activity under conditions relevant to winemaking (10 to 20°C, pH 3.5, 14% [vol/vol] ethanol). Kinetic constants were determined for EstB28 with p-nitrophenyl (pNP)-linked substrates ranging in chain length from C2 to C18. EstB28 exhibited greatest specificity for C2 to C4 pNP-linked substrates.

Characterization of a new virulent phage infecting the lactic acid bacterium Oenococcus oeni

2016 ◽  
Vol 54 ◽  
pp. 167-177 ◽  
Author(s):  
Fety Jaomanjaka ◽  
Olivier Claisse ◽  
Mélanie Blanche-Barbat ◽  
Melina Petrel ◽  
Patricia Ballestra ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Oenococcus Oeni ◽  
Virulent Phage

scholarly journals A partial proteome reference map of the wine lactic acid bacterium Oenococcus oeni ATCC BAA-1163

Open Biology ◽  
2014 ◽  
Vol 4 (2) ◽  
pp. 130154 ◽  
Author(s):  
María de la Luz Mohedano ◽  
Pasquale Russo ◽  
Vivian de los Ríos ◽  
Vittorio Capozzi ◽  
Pilar Fernández de Palencia ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Metabolic Pathways ◽  
Oenococcus Oeni ◽  
Transport Systems ◽  
Red Wines ◽  
Two Dimensional Gel Electrophoresis ◽  
Reference Map ◽  
Metabolic Activities

Oenococcus oeni is the main lactic acid bacterium that carries out the malolactic fermentation in virtually all red wines and in some white and sparkling wines. Oenococcus oeni possesses an array of metabolic activities that can modify the taste and aromatic properties of wine. There is, therefore, industrial interest in the proteins involved in these metabolic pathways and related transport systems of this bacterium. In this work, we report the characterization of the O. oeni ATCC BAA-1163 proteome. Total and membrane protein preparations from O. oeni were standardized and analysed by two-dimensional gel electrophoresis. Using tandem mass spectrometry, we identified 224 different spots corresponding to 152 unique proteins, which have been classified by their putative function and subjected to bioinformatics analysis.

Molecular Cloning, Heterologous Expression, and Characterization of Ornithine Decarboxylase from Oenococcus oeni

2011 ◽  
Vol 74 (8) ◽  
pp. 1309-1314 ◽  
Author(s):  
MARYSE BONNIN-JUSSERAND ◽  
COSETTE GRANDVALET ◽  
VANESSA DAVID ◽  
HERVÉ ALEXANDRE
Keyword(s):  
Lactic Acid ◽  
Heterologous Expression ◽  
Lactic Acid Bacterium ◽  
Ornithine Decarboxylase ◽  
Kinetic Studies ◽  
Maximal Activity ◽  
Oenococcus Oeni ◽  
Km Value ◽  
Oeni Strain

Ornithine decarboxylase (ODC) is responsible for the production of putrescine, the major biogenic amine found in wine. Oenococcus oeni is the most important lactic acid bacterium in the winemaking process and is involved in malolactic fermentation. We report here the characterization of ODC from an O. oeni strain isolated from wine. Screening of 263 strains isolated from wine and cider from all over the world revealed that the presence of the odc gene appears to be strain specific in O. oeni. After cloning, heterologous expression in Escherichia coli, and characterization, the enzyme was found to have a molecular mass of 85 kDa and a pI of 6.2 and revealed maximal activity at pH 5.5 and an optimum temperature of 35°C. Kinetic studies showed that O. oeni ODC is specific for l-ornithine with a Km value of 1 mM and a Vmax of 0.57 U·mg−1. The hypothesis that cadaverine, which results from lysine decarboxylation, may be linked to putrescine production is not valid since O. oeni ODC cannot decarboxylate L-lysine. As no lysine decarboxylase was detected in any of the O. oeni genomes sequenced, cadaverine synthesis may result from another metabolic pathway. This work is the first characterization of an ODC from a lactic acid bacterium isolated from a fermented product.

Cloning and functional characterization of an uncoupling protein homolog in hummingbirds

2001 ◽  
Vol 5 (3) ◽  
pp. 137-145 ◽  
Author(s):  
CLAUDIA R. VIANNA ◽  
THILO HAGEN ◽  
CHEN-YU ZHANG ◽  
ERIC BACHMAN ◽  
OLIVIER BOSS ◽  
...  
Keyword(s):  
Uncoupling Protein ◽  
Expression System ◽  
Functional Characterization ◽  
Pectoral Muscle ◽  
Mitochondrial Fraction ◽  
Mrna Levels ◽  
Coding Region ◽  
Reading Frame ◽  
Rapid Amplification

The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55% identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room’s temperature to 12–14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.

scholarly journals Characterization of γ-Aminobutyric acid(GABA) produced by a lactic acid bacterium from button mushroom bed

2013 ◽  
Vol 11 (4) ◽  
pp. 181-186 ◽  
Author(s):  
Yun-Seok Lee ◽  
Tae-Young Song ◽  
Won-Sik Kong ◽  
Min-Ho Yoon
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Aminobutyric Acid ◽  
Button Mushroom

scholarly journals Analysis of Transcriptomic Response to SO 2 by Oenococcus oeni Growing in Continuous Culture

2021 ◽  
Author(s):  
Cristobal A. Onetto ◽  
Peter J. Costello ◽  
Radka Kolouchova ◽  
Charlotte Jordans ◽  
Jane McCarthy ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
High Sensitivity ◽  
Malolactic Fermentation ◽  
Oenococcus Oeni ◽  
Low Ph ◽  
Transcriptomic Response ◽  
Content Type ◽  
Bacterium Species ◽  
High Ethanol

Malolactic fermentation is an indispensable step in the elaboration of most wines and is generally performed by Oenococcus oeni , a Gram-positive heterofermentative lactic acid bacterium species. While O. oeni is tolerant to many of the wine stresses, including low pH and high ethanol concentrations, it has high sensitivity to SO 2 , an antiseptic and antioxidant compound regularly used in winemaking.

scholarly journals Characterization of antifungal metabolites produced by novel lactic acid bacterium and their potential application as food biopreservatives

2019 ◽  
Vol 64 (1) ◽  
pp. 71-78 ◽  
Author(s):  
Mohamed G. Shehata ◽  
Ahmed N. Badr ◽  
Sobhy A. El Sohaimy ◽  
Dalal Asker ◽  
Tarek S. Awad
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Potential Application ◽  
Antifungal Metabolites

scholarly journals The Antisense RNA Approach: a New Application forIn VivoInvestigation of the Stress Response of Oenococcus oeni, a Wine-Associated Lactic Acid Bacterium

2015 ◽  
Vol 82 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Maud Darsonval ◽  
Tarek Msadek ◽  
Hervé Alexandre ◽  
Cosette Grandvalet
Keyword(s):  
Gene Expression ◽  
Lactic Acid ◽  
Stress Response ◽  
Lactic Acid Bacterium ◽  
Antisense Rna ◽  
Expression Vector ◽  
Stress Proteins ◽  
Oenococcus Oeni ◽  
Content Type

ABSTRACTOenococcus oeniis a wine-associated lactic acid bacterium mostly responsible for malolactic fermentation in wine. In wine,O. oenigrows in an environment hostile to bacterial growth (low pH, low temperature, and ethanol) that induces stress response mechanisms. To survive,O. oeniis known to set up transitional stress response mechanisms through the synthesis of heat stress proteins (HSPs) encoded by thehspgenes, notably a unique small HSP named Lo18. Despite the availability of the genome sequence, characterization ofO. oenigenes is limited, and little is known about thein vivorole of Lo18. Due to the lack of genetic tools forO. oeni, an efficient expression vector inO. oeniis still lacking, and deletion or inactivation of thehsp18gene is not presently practicable. As an alternative approach, with the goal of understanding the biological function of theO. oenihsp18genein vivo, we have developed an expression vector to produce antisense RNA targeting ofhsp18mRNA. Recombinant strains were exposed to multiple stresses inducinghsp18gene expression: heat shock and acid shock. We showed that antisense attenuation ofhsp18affectsO. oenisurvival under stress conditions. These results confirm the involvement of Lo18 in heat and acid tolerance ofO. oeni. Results of anisotropy experiments also confirm a membrane-protective role for Lo18, as previous observations had already suggested. This study describes a new, efficient tool to demonstrate the use of antisense technology for modulating gene expression inO. oeni.

Sign in / Sign up

Export Citation Format

Share Document