Characterization of an arylesterase from Lactobacillus helveticus CNRZ32

Fenster, Kurt M.; Parkin, Kirk L.; Steele, J. L.

doi:10.1046/j.1365-2672.2000.00993.x

Cited by 49 publications

(74 citation statements)

References 51 publications

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“…The gene estA encoding an esterase that can catalyze the biosynthesis of esters derived from short-chain fatty acids was cloned and characterized in lactococcal strains (28), as well as in Lactobacillus helveticus (26) and L. casei (74). These estA genes shared high sequence similarity and could be easily distinguished from other esterases (17,26; data not shown). Many LAB genomes, like those of the lactococci and streptococci, have one estA gene (Table 1).…”

Section: Enzymes Involved In the Transamination Routementioning

confidence: 99%

Comparative Genomics of Enzymes in Flavor-Forming Pathways from Amino Acids in Lactic Acid Bacteria

Liu

Nauta

Francke

et al. 2008

Appl Environ Microbiol

191

159

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4Lactic acid bacteria (LAB) have been widely used as starter or nonstarter cultures in the dairy industry for over a thousand years. They play an essential role in flavor formation during the fermentation of dairy products. Several metabolic routes can lead to the formation of flavor compounds when LAB are growing in milk. One of the main precursors for flavor compounds in milk is casein, although they can also be derived from fatty acids and sugars. The proteolytic system of LAB degrades casein into its constituent amino acids, which can be converted to flavor compounds. Although amino acid catabolism by LAB has been well researched (5, 31, 65, 78), many flavor-forming routes are yet to be discovered.Over 20 LAB genomes have been fully sequenced (1,11,12,16,19,41,44,45,53, 68,71). The available genomic information provides us with new opportunities to study the flavorforming potential of LAB. However, one of the main problems that one encounters while reconstructing flavor-forming routes based on the genomic information stored in the public databases is the inconsistency in the functional annotation for many of the relevant genes. These genes are mostly members of larger protein families. Moreover, even when the functional annotation in databases is appropriate, it sometimes reflects only part of the protein's full functional potential, since broad substrate specificities are often not taken into consideration in the annotation.We have now improved the functional annotation of the key enzymes in the formation of flavor compounds from amino acids by applying comparative genomics approaches that have been developed within our group to specify the annotation of homologous proteins, by combining phylogeny, gene context, and experimental evidence (32). We focused especially on the enzymes involved in the metabolism of sulfur-containing amino acids since these are known to be precursors of many flavor compounds in dairy fermentations. Comparative analysis of the various sequenced LAB species and strains resulted in an overall view of differences in their flavor-forming capacities.

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Section: Enzymes Involved In the Transamination Routementioning

confidence: 99%

Comparative Genomics of Enzymes in Flavor-Forming Pathways from Amino Acids in Lactic Acid Bacteria

Liu

Nauta

Francke

et al. 2008

Appl Environ Microbiol

191

159

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“…The wild-type MVP1 GDGL motif was mutated to EDGL in the mvp1 mutant. The Ser residue contained in the GDSL motif is crucial for enzymatic activity (Akoh et al, 2004), and multiple site-directed mutagenesis studies of lipases/esterases from Aeromonas hydrophila (Hilton and Buckley, 1991), Lactobacillus helveticus (Fenster et al, 2000), and Vibrio mimicus (Shaw et al, 1994) have demonstrated that mutagenesis of the Ser residue in the GDSL motif completely abolishes enzymatic activity. We found that MVP1 did in fact lack lipase activity.…”

Section: Cloning and Identification Of Mvp1 As A Myapmentioning

confidence: 99%

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

et al. 2009

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We identified an Arabidopsis (Arabidopsis thaliana) ethyl methanesulfonate mutant, modified vacuole phenotype1-1 (mvp1-1), in a fluorescent confocal microscopy screen for plants with mislocalization of a green fluorescent protein-d tonoplast intrinsic protein fusion. The mvp1-1 mutant displayed static perinuclear aggregates of the reporter protein. mvp1 mutants also exhibited a number of vacuole-related phenotypes, as demonstrated by defects in growth, utilization of stored carbon, gravitropic response, salt sensitivity, and specific susceptibility to the fungal necrotroph Alternaria brassicicola. Similarly, crosses with other endomembrane marker fusions identified mislocalization to aggregate structures, indicating a general defect in protein trafficking. Map-based cloning showed that the mvp1-1 mutation altered a gene encoding a putative myrosinase-associated protein, and glutathione S-transferase pull-down assays demonstrated that MVP1 interacted specifically with the Arabidopsis myrosinase protein, THIOGLUCOSIDE GLUCOHYDROLASE2 (TGG2), but not TGG1. Moreover, the mvp1-1 mutant showed increased nitrile production during glucosinolate hydrolysis, suggesting that MVP1 may play a role in modulation of myrosinase activity. We propose that MVP1 is a myrosinase-associated protein that functions, in part, to correctly localize the myrosinase TGG2 and prevent inappropriate glucosinolate hydrolysis that could generate cytotoxic molecules.

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“…Most characterization of esterases in LAB has focused on dairy isolates (9,(16)(17)(18)20). Parallel work in a wine context is limited despite general acceptance of the importance of esters in wine.…”

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confidence: 99%

Cloning and Characterization of an Intracellular Esterase from the Wine-Associated Lactic Acid BacteriumOenococcus oeni

Sumby

Matthews

Grbin

et al. 2009

Appl Environ Microbiol

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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 C 2 to C 18 . EstB28 exhibited greatest specificity for C 2 to C 4 pNP-linked substrates.

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Characterization of an arylesterase from Lactobacillus helveticus CNRZ32

Cited by 49 publications

References 51 publications

Comparative Genomics of Enzymes in Flavor-Forming Pathways from Amino Acids in Lactic Acid Bacteria

Comparative Genomics of Enzymes in Flavor-Forming Pathways from Amino Acids in Lactic Acid Bacteria

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

Cloning and Characterization of an Intracellular Esterase from the Wine-Associated Lactic Acid BacteriumOenococcus oeni

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