Munoz R

References (24)

Title : A new and promiscuous alpha\/beta hydrolase from Acinetobacter tandoii DSM 14970( T) inactivates the mycotoxin ochratoxin A - Sanchez-Arroyo_2024_Appl.Microbiol.Biotechnol_108_230
Author(s) : Sanchez-Arroyo A , Plaza-Vinuesa L , Abeijon-Mukdsi MC , de Las Rivas B , Mancheno JM , Munoz R
Ref : Applied Microbiology & Biotechnology , 108 :230 , 2024
Abstract : The presence of ochratoxin A (OTA) in food and feed represents a serious concern since it raises severe health implications. Bacterial strains of the Acinetobacter genus hydrolyse the amide bond of OTA yielding non-toxic OTalpha and L-beta-phenylalanine; in particular, the carboxypeptidase PJ15_1540 from Acinetobacter sp. neg1 has been identified as an OTA-degrading enzyme. Here, we describe the ability to transform OTA of cell-free protein extracts from Acinetobacter tandoii DSM 14970( T), a strain isolated from sludge plants, and also report on the finding of a new and promiscuous alpha/beta hydrolase (ABH), with close homologs highly distributed within the Acinetobacter genus. ABH from A. tandoii (AtABH) exhibited amidase activity against OTA and OTB mycotoxins, as well as against several carboxypeptidase substrates. The predicted structure of AtABH reveals an alpha/beta hydrolase core composed of a parallel, six-stranded beta-sheet, with a large cap domain similar to the marine esterase EprEst. Further biochemical analyses of AtABH reveal that it is an efficient esterase with a similar specificity profile as EprEst. Molecular docking studies rendered a consistent OTA-binding mode. We proposed a potential procedure for preparing new OTA-degrading enzymes starting from promiscuous alpha/beta hydrolases based on our results. KEY POINTS: AtABH is a promiscuous alphabeta hydrolase with both esterase and amidohydrolase activities AtABH hydrolyses the amide bond of ochratoxin A rendering nontoxic OTalpha Promiscuous alphabeta hydrolases are a possible source of new OTA-degrading enzymes.
ESTHER : Sanchez-Arroyo_2024_Appl.Microbiol.Biotechnol_108_230
PubMedSearch : Sanchez-Arroyo_2024_Appl.Microbiol.Biotechnol_108_230
PubMedID: 38393350
Gene_locus related to this paper: 9gamm-acitaABH

Title : Screening Enzymes That Can Depolymerize Commercial Biodegradable Polymers: Heterologous Expression of Fusarium solani Cutinase in Escherichia coli - Santos-Beneit_2023_Microorganisms_11_
Author(s) : Santos-Beneit F , Chen LM , Bordel S , Frutos de la Flor R , Garcia-Depraect O , Lebrero R , Rodriguez-Vega S , Munoz R , Borner RA , Borner T
Ref : Microorganisms , 11 : , 2023
Abstract : In recent years, a number of microbial enzymes capable of degrading plastics have been identified. Biocatalytic depolymerization mediated by enzymes has emerged as a potentially more efficient and environmentally friendly alternative to the currently employed methods for plastic treatment and recycling. However, the functional and systematic study of depolymerase enzymes with respect to the degradation of a series of plastic polymers in a single work has not been widely addressed at present. In this study, the ability of a set of enzymes (esterase, arylesterase and cutinase) to degrade commercial biodegradable polymers (PBS, PBAT, PHB, PHBH, PHBV, PCL, PLA and PLA/PCL) and the effect of pre-treatment methods on their degradation rate was assessed. The degradation products were identified and quantified by HPLC and LC-HRMS analysis. Out of the three enzymes, Fusarium solani cutinase (FsCut) showed the highest activity on grinded PBAT, PBS and PCL after 7 days of incubation. FsCut was engineered and heterologous expressed in Escherichia coli, which conferred the bacterium the capability of degrading solid discs of PBAT and to grow in PBS as the sole carbon source of the medium.
ESTHER : Santos-Beneit_2023_Microorganisms_11_
PubMedSearch : Santos-Beneit_2023_Microorganisms_11_
PubMedID: 36838293

Title : A structurally unique Fusobacterium nucleatum tannase provides detoxicant activity against gallotannins and pathogen resistance - Mancheno_2022_Microb.Biotechnol_15_648
Author(s) : Mancheno JM , Atondo E , Tomas-Cortazar J , Luis Lavin J , Plaza-Vinuesa L , Martin-Ruiz I , Barriales D , Palacios A , Daniel Navo C , Sampedro L , Pena-Cearra A , Angel Pascual-Itoiz M , Castelo J , Carreras-Gonzalez A , Castellana D , Pellon A , Delgado S , Ruas-Madiedo P , de Las Rivas B , Abecia L , Munoz R , Jimenez-Oses G , Anguita J , Rodriguez H
Ref : Microb Biotechnol , 15 :648 , 2022
Abstract : Colorectal cancer pathogenesis and progression is associated with the presence of Fusobacterium nucleatum and the reduction of acetylated derivatives of spermidine, as well as dietary components such as tannin-rich foods. We show that a new tannase orthologue of F. nucleatum (TanBF(nn) ) has significant structural differences with its Lactobacillus plantarum counterpart affecting the flap covering the active site and the accessibility of substrates. Crystallographic and molecular dynamics analysis revealed binding of polyamines to a small cavity that connects the active site with the bulk solvent which interact with catalytically indispensable residues. As a result, spermidine and its derivatives, particularly N(8) -acetylated spermidine, inhibit the hydrolytic activity of TanBF(nn) and increase the toxicity of gallotannins to F. nucleatum. Our results support a model in which the balance between the detoxicant activity of TanBF(nn) and the presence of metabolic inhibitors can dictate either conducive or unfavourable conditions for the survival of F. nucleatum.
ESTHER : Mancheno_2022_Microb.Biotechnol_15_648
PubMedSearch : Mancheno_2022_Microb.Biotechnol_15_648
PubMedID: 33336898
Gene_locus related to this paper: fusnn-FN0616

Title : The use of Lactobacillus plantarum esterase genes: a biotechnological strategy to increase the bioavailability of dietary phenolic compounds in lactic acid bacteria - Landete_2021_Int.J.Food.Sci.Nutr__1
Author(s) : Landete JM , Plaza-Vinuesa L , Montenegro C , Santamaria L , Reveron I , de Las Rivas B , Munoz R
Ref : Int J Food Sci Nutr , :1 , 2021
Abstract : In Lactobacillus plantarum the metabolism of hydroxybenzoic and hydroxycinnamic acid derivatives follows a similar two-step pathway, an esterase action followed by a decarboxylation. The L. plantarum esterase genes involved in these reactions have been cloned into pNZ8048 or pT1NX plasmids and transformed into technologically relevant lactic acid bacteria. None of the strains assayed can hydrolyse methyl gallate, a hydroxybenzoic ester. The presence of the L. plantarum tannase encoding genes (tanA(Lp) or tanB(Lp)) on these bacteria conferred their detectable esterase (tannase) activity. Similarly, on hydroxycinnamic compounds, esterase activity for the hydrolysis of ferulic acid was acquired by lactic acid bacteria when L. plantarum esterase (JDM1_1092) was present. This study showed that the heterologous expression of L. plantarum esterase genes involved in the metabolism of phenolic acids allowed the production of healthy compounds which increase the bioavailability of these dietary compounds in food relevant lactic acid bacteria.
ESTHER : Landete_2021_Int.J.Food.Sci.Nutr__1
PubMedSearch : Landete_2021_Int.J.Food.Sci.Nutr__1
PubMedID: 33730985

Title : Bacterial tannases: classification and biochemical properties - de Las Rivas_2019_Appl.Microbiol.Biotechnol_103_603
Author(s) : de Las Rivas B , Rodriguez H , Anguita J , Munoz R
Ref : Applied Microbiology & Biotechnology , 103 :603 , 2019
Abstract : Tannin acyl hydrolases, also known as tannases, are a group of enzymes critical for the transformation of tannins. The study of these enzymes, which initially evolved in different organisms to detoxify and/or use these plant metabolites, has nowadays become relevant in microbial enzymology research due to their relevant role in food tannin transformation. Microorganisms, particularly bacteria, are major sources of tannase. Cloning and heterologous expression of bacterial tannase genes and structural studies have been performed in the last few years. However, a systematic compilation of the information related to all recombinant tannases, their classification, and characteristics is missing. In this review, we explore the diversity of heterologously produced bacterial tannases, describing their substrate specificity and biochemical characterization. Moreover, a new classification based on sequence similarity analysis is proposed. Finally, putative tannases have been identified in silico for each group of tannases taking advantage of the use of the "tannase" distinctive features previously proposed.
ESTHER : de Las Rivas_2019_Appl.Microbiol.Biotechnol_103_603
PubMedSearch : de Las Rivas_2019_Appl.Microbiol.Biotechnol_103_603
PubMedID: 30460533

Title : Identification of a highly active tannase enzyme from the oral pathogen Fusobacterium nucleatum subsp. polymorphum - Tomas-Cortazar_2018_Microb.Cell.Fact_17_33
Author(s) : Tomas-Cortazar J , Plaza-Vinuesa L , de Las Rivas B , Lavin JL , Barriales D , Abecia L , Mancheno JM , Aransay AM , Munoz R , Anguita J , Rodriguez H
Ref : Microb Cell Fact , 17 :33 , 2018
Abstract : BACKGROUND: Tannases are tannin-degrading enzymes that have been described in fungi and bacteria as an adaptative mechanism to overcome the stress conditions associated with the presence of these phenolic compounds. RESULTS: We have identified and expressed in E. coli a tannase from the oral microbiota member Fusobacterium nucleatum subs. polymorphum (TanBFnp). TanBFnp is the first tannase identified in an oral pathogen. Sequence analyses revealed that it is closely related to other bacterial tannases. The enzyme exhibits biochemical properties that make it an interesting target for industrial use. TanBFnp has one of the highest specific activities of all bacterial tannases described to date and shows optimal biochemical properties such as a high thermal stability: the enzyme keeps 100% of its activity after prolonged incubations at different temperatures up to 45 degrees C. TanBFnp also shows a wide temperature range of activity, maintaining above 80% of its maximum activity between 22 and 55 degrees C. The use of a panel of 27 esters of phenolic acids demonstrated activity of TanBFnp only against esters of gallic and protocatechuic acid, including tannic acid, gallocatechin gallate and epigallocatechin gallate. Overall, TanBFnp possesses biochemical properties that make the enzyme potentially useful in biotechnological applications. CONCLUSIONS: We have identified and characterized a metabolic enzyme from the oral pathogen Fusobacterium nucleatum subsp. polymorphum. The biochemical properties of TanBFnp suggest that it has a major role in the breakdown of complex food tannins during oral processing. Our results also provide some clues regarding its possible participation on bacterial survival in the oral cavity. Furthermore, the characteristics of this enzyme make it of potential interest for industrial use.
ESTHER : Tomas-Cortazar_2018_Microb.Cell.Fact_17_33
PubMedSearch : Tomas-Cortazar_2018_Microb.Cell.Fact_17_33
PubMedID: 29482557
Gene_locus related to this paper: fusnp-a5txs5

Title : Differential Gene Expression by Lactobacillus plantarum WCFS1 in Response to Phenolic Compounds Reveals New Genes Involved in Tannin Degradation - Reveron_2017_Appl.Environ.Microbiol_83_
Author(s) : Reveron I , Jimenez N , Curiel JA , Penas E , Lopez de Felipe F , de Las Rivas B , Munoz R
Ref : Applied Environmental Microbiology , 83 : , 2017
Abstract : Lactobacillus plantarum is a lactic acid bacterium that can degrade food tannins by the successive action of tannase and gallate decarboxylase enzymes. In the L. plantarum genome, the gene encoding the catalytic subunit of gallate decarboxylase (lpdC, or lp_2945) is only 6.5 kb distant from the gene encoding inducible tannase (L. plantarumtanB [tanB(Lp) ], or lp_2956). This genomic context suggests concomitant activity and regulation of both enzymatic activities. Reverse transcription analysis revealed that subunits B (lpdB, or lp_0271) and D (lpdD, or lp_0272) of the gallate decarboxylase are cotranscribed, whereas subunit C (lpdC, or lp_2945) is cotranscribed with a gene encoding a transport protein (gacP, or lp_2943). In contrast, the tannase gene is transcribed as a monocistronic mRNA. Investigation of knockout mutations of genes located in this chromosomal region indicated that only mutants of the gallate decarboxylase (subunits B and C), tannase, GacP transport protein, and TanR transcriptional regulator (lp_2942) genes exhibited altered tannin metabolism. The expression profile of genes involved in tannin metabolism was also analyzed in these mutants in the presence of methyl gallate and gallic acid. It is noteworthy that inactivation of tanR suppresses the induction of all genes overexpressed in the presence of methyl gallate and gallic acid. This transcriptional regulator was also induced in the presence of other phenolic compounds, such as kaempferol and myricetin. This study complements the catalog of L. plantarum expression profiles responsive to phenolic compounds, which enable this bacterium to adapt to a plant food environment.IMPORTANCELactobacillus plantarum is a bacterial species frequently found in the fermentation of vegetables when tannins are present. L. plantarum strains degrade tannins to the less-toxic pyrogallol by the successive action of tannase and gallate decarboxylase enzymes. The genes encoding these enzymes are located close to each other in the chromosome, suggesting concomitant regulation. Proteins involved in tannin metabolism and regulation, such GacP (gallic acid permease) and TanR (tannin transcriptional regulator), were identified by differential gene expression in knockout mutants with mutations in genes from this region. This study provides insights into the highly coordinated mechanisms that enable L. plantarum to adapt to plant food fermentations.
ESTHER : Reveron_2017_Appl.Environ.Microbiol_83_
PubMedSearch : Reveron_2017_Appl.Environ.Microbiol_83_
PubMedID: 28115379

Title : The Lp_3561 and Lp_3562 Enzymes Support a Functional Divergence Process in the Lipase\/Esterase Toolkit from Lactobacillus plantarum - Esteban-Torres_2016_Front.Microbiol_7_1118
Author(s) : Esteban-Torres M , Reveron I , Santamaria L , Mancheno JM , de Las Rivas B , Munoz R
Ref : Front Microbiol , 7 :1118 , 2016
Abstract : Lactobacillus plantarum species is a good source of esterases since both lipolytic and esterase activities have been described for strains of this species. No fundamental biochemical difference exists among esterases and lipases since both share a common catalytic mechanism. L. plantarum WCFS1 possesses a protein, Lp_3561, which is 44% identical to a previously described lipase, Lp_3562. In contrast to Lp_3562, Lp_3561 was unable to degrade esters possessing a chain length higher than C4 and the triglyceride tributyrin. As in other L. plantarum esterases, the electrostatic potential surface around the active site in Lp_3561 is predicted to be basic, whereas it is essentially neutral in the Lp_3562 lipase. The fact that the genes encoding both proteins were located contiguously in the L. plantarum WCFS1 genome, suggests that they originated by tandem duplication, and therefore are paralogs as new functions have arisen during evolution. The presence of the contiguous lp_3561 and lp_3562 genes was studied among L. plantarum strains. They are located in a 8,903 bp DNA fragment that encodes proteins involved in the catabolism of sialic acid and are predicted to increase bacterial adaptability under certain growth conditions.
ESTHER : Esteban-Torres_2016_Front.Microbiol_7_1118
PubMedSearch : Esteban-Torres_2016_Front.Microbiol_7_1118
PubMedID: 27486450
Gene_locus related to this paper: lacpl-LP.3561 , lacpl-LP.3562

Title : A Lactobacillus plantarum Esterase Active on a Broad Range of Phenolic Esters - Esteban-Torres_2015_Appl.Environ.Microbiol_81_3235
Author(s) : Esteban-Torres M , Landete JM , Reveron I , Santamaria L , de Las Rivas B , Munoz R
Ref : Applied Environmental Microbiology , 81 :3235 , 2015
Abstract : Lactobacillus plantarum is the lactic acid bacterial species most frequently found in the fermentation of food products of plant origin on which phenolic compounds are abundant. L. plantarum strains showed great flexibility in their ability to adapt to different environments and growth substrates. Of 28 L. plantarum strains analyzed, only cultures from 7 strains were able to hydrolyze hydroxycinnamic esters, such as methyl ferulate or methyl caffeate. As revealed by PCR, only these seven strains possessed the est_1092 gene. When the est_1092 gene was introduced into L. plantarum WCFS1 or L. lactis MG1363, their cultures acquired the ability to degrade hydroxycinnamic esters. These results support the suggestion that Est_1092 is the enzyme responsible for the degradation of hydroxycinnamic esters on the L. plantarum strains analyzed. The Est_1092 protein was recombinantly produced and biochemically characterized. Surprisingly, Est_1092 was able to hydrolyze not only hydroxycinnamic esters, since all the phenolic esters assayed were hydrolyzed. Quantitative PCR experiments revealed that the expression of est_1092 was induced in the presence of methyl ferulate, an hydroxycinnamic ester, but was inhibited on methyl gallate, an hydroxybenzoic ester. As Est_1092 is an enzyme active on a broad range of phenolic esters, simultaneously possessing feruloyl esterase and tannase activities, its presence on some L. plantarum strains provides them with additional advantages to survive and grow on plant environments.
ESTHER : Esteban-Torres_2015_Appl.Environ.Microbiol_81_3235
PubMedSearch : Esteban-Torres_2015_Appl.Environ.Microbiol_81_3235
PubMedID: 25746986
Gene_locus related to this paper: lacpl-Est.1092

Title : Characterisation of a cold-active and salt-tolerant esterase from Lactobacillus plantarum with potential application during cheese ripening - Esteban-Torres_2014_Int.Dairy.J_39_312
Author(s) : Esteban-Torres M , Santamaria L , de Las Rivas B , Munoz R
Ref : International Dairy Journal , 39 :312 , 2014
Abstract : During cheese ripening, the metabolic activity of microorganisms influences cheese flavour. Lactobacillus plantarum is a non-starter lactic acid bacterium that can be found during cheese ripening. An esterase-encoding gene from L. plantarum, lp_3505, was cloned and expressed in Escherichia coli BL21 (DE3). The biotechnologically produced Lp_3505 protein was purified as an active soluble form using His-tag affinity chromatography. The enzyme has an optimal pH and temperature of 6 and 5 C, respectively. The enzyme showed remarkable stability at 20 C, retaining more than 60% of its maximal activity after 20 h incubation at this temperature. In addition, NaCl concentrations lower than 20% increased esterase activity. The cold-activity and salt-tolerance exhibited by Lp_3505 indicated that this esterase could be a useful exogenous enzyme to be added for cheese ripening.
ESTHER : Esteban-Torres_2014_Int.Dairy.J_39_312
PubMedSearch : Esteban-Torres_2014_Int.Dairy.J_39_312
Gene_locus related to this paper: lacpl-EST2

Title : Esterase LpEst1 from Lactobacillus plantarum: A Novel and Atypical Member of the alphabeta Hydrolase Superfamily of Enzymes - Alvarez_2014_PLoS.One_9_e92257
Author(s) : Alvarez Y , Esteban-Torres M , Cortes-Cabrera A , Gago F , Acebron I , Benavente R , Mardo K , de Las Rivas B , Munoz R , Mancheno JM
Ref : PLoS ONE , 9 :e92257 , 2014
Abstract : The genome of the lactic acid bacterium Lactobacillus plantarum WCFS1 reveals the presence of a rich repertoire of esterases and lipases highlighting their important role in cellular metabolism. Among them is the carboxylesterase LpEst1 a bacterial enzyme related to the mammalian hormone-sensitive lipase, which is known to play a central role in energy homeostasis. In this study, the crystal structure of LpEst1 has been determined at 2.05 A resolution; it exhibits an alphabeta-hydrolase fold, consisting of a central beta-sheet surrounded by alpha-helices, endowed with novel topological features. The structure reveals a dimeric assembly not comparable with any other enzyme from the bacterial hormone-sensitive lipase family, probably echoing the specific structural features of the participating subunits. Biophysical studies including analytical gel filtration and ultracentrifugation support the dimeric nature of LpEst1. Structural and mutational analyses of the substrate-binding pocket and active site together with biochemical studies provided insights for understanding the substrate profile of LpEst1 and suggested for the first time the conserved Asp173, which is adjacent to the nucleophile, as a key element in the stabilization of the loop where the oxyanion hole resides.
ESTHER : Alvarez_2014_PLoS.One_9_e92257
PubMedSearch : Alvarez_2014_PLoS.One_9_e92257
PubMedID: 24663330
Gene_locus related to this paper: lacpl-LP.0973

Title : Characterization of a versatile arylesterase from Lactobacillus plantarum active on wine esters - Esteban-Torres_2014_J.Agric.Food.Chem_62_5118
Author(s) : Esteban-Torres M , Barcenilla JM , Mancheno JM , de Las Rivas B , Munoz R
Ref : Journal of Agricultural and Food Chemistry , 62 :5118 , 2014
Abstract : The gene lp_1002 from Lactobacillus plantarum WCFS1 encoding a putative lipase/esterase was cloned and overexpressed in Escherichia coli BL21(DE3). The purified Lp_1002 protein was biochemically characterized. Lp_1002 is an arylesterase which showed high hydrolytic activity on phenyl acetate. Although to a lesser extent, Lp_1002 also hydrolyzed most of the esters assayed including relevant wine aroma compounds. Importantly, Lp_1002 exhibited hydrolytic activity at winemaking conditions, although optimal catalytic activity is observed at 40 degreesC and pH 5-7. The effect of wine compounds on Lp_1002 activity was assayed. From the compounds assayed (ethanol, sodium metabisulfite, and malic, tartaric, lactic and citric acids), only malic acid slightly inhibited Lp_1002 activity. Lp_1002 is the first arylesterase described in a wine lactic acid bacteria and possessed suitable biochemical properties to be used during winemaking.
ESTHER : Esteban-Torres_2014_J.Agric.Food.Chem_62_5118
PubMedSearch : Esteban-Torres_2014_J.Agric.Food.Chem_62_5118
PubMedID: 24856385
Gene_locus related to this paper: lacpl-LP.1002

Title : Production and characterization of a tributyrin esterase from Lactobacillus plantarum suitable for cheese lipolysis - Esteban-Torres_2014_J.Dairy.Sci_97_6737
Author(s) : Esteban-Torres M , Mancheno JM , de Las Rivas B , Munoz R
Ref : J Dairy Sci , 97 :6737 , 2014
Abstract : Lactobacillus plantarum is a lactic acid bacterium that can be found during cheese ripening. Lipolysis of milk triacylglycerols to free fatty acids during cheese ripening has fundamental consequences on cheese flavor. In the present study, the gene lp_1760, encoding a putative esterase or lipase, was cloned and expressed in Escherichia coli BL21 (DE3) and the overproduced Lp_1760 protein was biochemically characterized. Lp_1760 hydrolyzed p-nitrophenyl esters of fatty acids from C2 to C16, with a preference for p-nitrophenyl butyrate. On triglycerides, Lp_1760 showed higher activity on tributyrin than on triacetin. Although optimal conditions for activity were 45 degreesC and pH 7, Lp_1760 retains activity under conditions commonly found during cheese making and ripening. The Lp_1760 showed more than 50% activity at 5 degreesC and exhibited thermal stability at high temperatures. Enzymatic activity was strongly inhibited by sodium dodecyl sulfate and phenylmethylsulfonyl fluoride. The Lp_1760 tributyrin esterase showed high activity in the presence of NaCl, lactic acid, and calcium chloride. The results suggest that Lp_1760 might be a useful tributyrin esterase to be used in cheese manufacturing.
ESTHER : Esteban-Torres_2014_J.Dairy.Sci_97_6737
PubMedSearch : Esteban-Torres_2014_J.Dairy.Sci_97_6737
PubMedID: 25173466
Gene_locus related to this paper: lacpl-LP.1760

Title : Characterization of a cold-active esterase from Lactobacillus plantarum suitable for food fermentations - Esteban-Torres_2014_J.Agric.Food.Chem_62_5126
Author(s) : Esteban-Torres M , Mancheno JM , de Las Rivas B , Munoz R
Ref : Journal of Agricultural and Food Chemistry , 62 :5126 , 2014
Abstract : Lactobacillus plantarum is a lactic acid bacteria that can be found in numerous fermented foods. Esterases from L. plantarum exert a fundamental role in food aroma. In the present study, the gene lp_2631 encoding a putative esterase was cloned and expressed in Escherichia coli BL21 (DE3) and the overproduced Lp_2631 protein has been biochemically characterized. Lp_2631 exhibited optimal esterase activity at 20 degreesC and more than 90% of maximal activity at 5 degreesC, being the first cold-active esterase described in a lactic acid bacteria. Lp_2631 exhibited 40% of its maximal activity after 2 h of incubation at 65 degreesC. Lp_2631 also showed marked activity in the presence of compounds commonly found in food fermentations, such as NaCl, ethanol, or lactic acid. The results suggest that Lp_2631 might be a useful esterase to be used in food fermentations.
ESTHER : Esteban-Torres_2014_J.Agric.Food.Chem_62_5126
PubMedSearch : Esteban-Torres_2014_J.Agric.Food.Chem_62_5126
PubMedID: 24856291
Gene_locus related to this paper: lacpl-LP.2631

Title : Tannin degradation by a novel tannase enzyme present in some Lactobacillus plantarum strains - Jimenez_2014_Appl.Environ.Microbiol_80_2991
Author(s) : Jimenez N , Esteban-Torres M , Mancheno JM , de Las Rivas B , Munoz R
Ref : Applied Environmental Microbiology , 80 :2991 , 2014
Abstract : Lactobacillus plantarum is frequently isolated from the fermentation of plant material where tannins are abundant. L. plantarum strains possess tannase activity to degrade plant tannins. An L. plantarum tannase (TanBLp, formerly called TanLp1) was previously identified and biochemically characterized. In this study, we report the identification and characterization of a novel tannase (TanALp). While all 29 L. plantarum strains analyzed in the study possess the tanBLp gene, the gene tanALp was present in only four strains. Upon methyl gallate exposure, the expression of tanBLp was induced, whereas tanALp expression was not affected. TanALp showed only 27% sequence identity to TanBLp, but the residues involved in tannase activity are conserved. Optimum activity for TanALp was observed at 30 degreesC and pH 6 in the presence of Ca(2+) ions. TanALp was able to hydrolyze gallate and protocatechuate esters with a short aliphatic alcohol substituent. Moreover, TanALp was able to fully hydrolyze complex gallotannins, such as tannic acid. The presence of the extracellular TanALp tannase in some L. plantarum strains provides them an advantage for the initial degradation of complex tannins present in plant environments.
ESTHER : Jimenez_2014_Appl.Environ.Microbiol_80_2991
PubMedSearch : Jimenez_2014_Appl.Environ.Microbiol_80_2991
PubMedID: 24610854
Gene_locus related to this paper: lacpn-d7vbf4

Title : Genetic and biochemical approaches towards unravelling the degradation of gallotannins by Streptococcus gallolyticus - Jimenez_2014_Microb.Cell.Fact_13_154
Author(s) : Jimenez N , Reveron I , Esteban-Torres M , Lopez de Felipe F , de Las Rivas B , Munoz R
Ref : Microb Cell Fact , 13 :154 , 2014
Abstract : BACKGROUND: Herbivores have developed mechanisms to overcome adverse effects of dietary tannins through the presence of tannin-resistant bacteria. Tannin degradation is an unusual characteristic among bacteria. Streptococcus gallolyticus is a common tannin-degrader inhabitant of the gut of herbivores where plant tannins are abundant. The biochemical pathway for tannin degradation followed by S. gallolyticus implies the action of tannase and gallate decarboxylase enzymes to produce pyrogallol, as final product. From these proteins, only a tannase (TanBSg) has been characterized so far, remaining still unknown relevant proteins involved in the degradation of tannins. RESULTS: In addition to TanBSg, genome analysis of S. gallolyticus subsp. gallolyticus strains revealed the presence of an additional protein similar to tannases, TanASg (GALLO_0933). Interestingly, this analysis also indicated that only S. gallolyticus strains belonging to the subspecies "gallolyticus" possessed tannase copies. This observation was confirmed by PCR on representative strains from different subspecies. In S. gallolyticus subsp. gallolyticus the genes encoding gallate decarboxylase are clustered together and close to TanBSg, however, TanASg is not located in the vicinity of other genes involved in tannin metabolism. The expression of the genes enconding gallate decarboxylase and the two tannases was induced upon methyl gallate exposure. As TanBSg has been previously characterized, in this work the tannase activity of TanASg was demonstrated in presence of phenolic acid esters. TanASg showed optimum activity at pH 6.0 and 37 degreesC. As compared to the tannin-degrader Lactobacillus plantarum strains, S. gallolyticus presented several advantages for tannin degradation. Most of the L. plantarum strains possessed only one tannase enzyme (TanBLp), whereas all the S. gallolytcius subsp. gallolyticus strains analyzed possesses both TanASg and TanBSg proteins. More interestingly, upon methyl gallate induction, only the tanB Lp gene was induced from the L. plantarum tannases; in contrast, both tannase genes were highly induced in S. gallolyticus. Finally, both S. gallolyticus tannase proteins presented higher activity than their L. plantarum counterparts. CONCLUSIONS: The specific features showed by S. gallolyticus subsp. gallolyticus in relation to tannin degradation indicated that strains from this subspecies could be considered so far the best bacterial cellular factories for tannin degradation.
ESTHER : Jimenez_2014_Microb.Cell.Fact_13_154
PubMedSearch : Jimenez_2014_Microb.Cell.Fact_13_154
PubMedID: 25359406
Gene_locus related to this paper: strg3-d3hd15 , strg3-d3hey7

Title : Structure, biochemical characterization and analysis of the pleomorphism of carboxylesterase Cest-2923 from Lactobacillus plantarum WCFS1 - Benavente_2013_FEBS.J_280_6658
Author(s) : Benavente R , Esteban-Torres M , Acebron I , de Las Rivas B , Munoz R , Alvarez Y , Mancheno JM
Ref : Febs J , 280 :6658 , 2013
Abstract : The hydrolase fold is one of the most versatile structures in the protein realm according to the diversity of sequences adopting such a three-dimensional architecture. In the present study, we clarified the crystal structure of the carboxylesterase Cest-2923 from the lactic acid bacterium Lactobacillus plantarum WCFS1 refined to 2.1 A resolution, determined its main biochemical characteristics and also carried out an analysis of its associative behaviour in solution. We found that the versatility of a canonical alpha/beta hydrolase fold, the basic framework of the crystal structure of Cest-2923, also extends to its oligomeric behaviour in solution. Thus, we discovered that Cest-2923 exhibits a pH-dependent pleomorphic behaviour in solution involving monomers, canonical dimers and tetramers. Although, at neutral pH, the system is mainly shifted to dimeric species, under acidic conditions, tetrameric species predominate. Despite these tetramers resulting from the association of canonical dimers, as is commonly found in many other carboxylesterases from the hormone-sensitive lipase family, they can be defined as 'noncanonical' because they represent a different association mode. We identified this same type of tetramer in the closest relative of Cest-2923 that has been structurally characterized: the sugar hydrolase YeeB from Lactococcus lactis. The observed associative behaviour is consistent with the different crystallographic results for Cest-2923 from structural genomics consortia. Finally, the presence of sulfate or acetate molecules (depending on the crystal form analysed) in the close vicinity of the nucleophile Ser116 allows us to identify interactions with the putative oxyanion hole and deduce the existence of hydrolytic activity within Cest-2923 crystals. STRUCTURED DIGITAL ABSTRACT: Cest-2923 and Cest-2923 bind by x-ray crystallography (1, 2) Cest-2923 and Cest-2923 bind by cosedimentation in solution (1, 2) DATABASE: The atomic coordinates and structure factors have been deposited in the Protein Data Bank with accession numbers: 4BZW for Cest-2923 from native crystals not soaked with substrates (P63 22 space group); 4C01 for Cest-2923 from crystals soaked with phenyl acetate (C2 space group); 4BZZ for Cest-2923 from crystals soaked with isopropenyl acetate (P622 space group).
ESTHER : Benavente_2013_FEBS.J_280_6658
PubMedSearch : Benavente_2013_FEBS.J_280_6658
PubMedID: 24127688
Gene_locus related to this paper: lacpl-LP.2923

Title : Characterization of a feruloyl esterase from Lactobacillus plantarum - Esteban-Torres_2013_Appl.Environ.Microbiol_79_5130
Author(s) : Esteban-Torres M , Reveron I , Mancheno JM , de Las Rivas B , Munoz R
Ref : Applied Environmental Microbiology , 79 :5130 , 2013
Abstract : Lactobacillus plantarum is frequently found in the fermentation of plant-derived food products, where hydroxycinnamoyl esters are abundant. L. plantarum WCFS1 cultures were unable to hydrolyze hydroxycinnamoyl esters; however, cell extracts from the strain partially hydrolyze methyl ferulate and methyl p-coumarate. In order to discover whether the protein Lp_0796 is the enzyme responsible for this hydrolytic activity, it was recombinantly overproduced and enzymatically characterized. Lp_0796 is an esterase that, among other substrates, is able to efficiently hydrolyze the four model substrates for feruloyl esterases (methyl ferulate, methyl caffeate, methyl p-coumarate, and methyl sinapinate). A screening test for the detection of the gene encoding feruloyl esterase Lp_0796 revealed that it is generally present among L. plantarum strains. The present study constitutes the description of feruloyl esterase activity in L. plantarum and provides new insights into the metabolism of hydroxycinnamic compounds in this bacterial species.
ESTHER : Esteban-Torres_2013_Appl.Environ.Microbiol_79_5130
PubMedSearch : Esteban-Torres_2013_Appl.Environ.Microbiol_79_5130
PubMedID: 23793626

Title : A novel automated flow-based biosensor for the determination of organophosphate pesticides in milk - Mishra_2012_Biosens.Bioelectron_32_56
Author(s) : Mishra RK , Dominguez RB , Bhand S , Munoz R , Marty JL
Ref : Biosensors & Bioelectronics , 32 :56 , 2012
Abstract : This work describes the development of an automated flow-based biosensor that employs genetically modified acetylcholinesterase (AChE) enzymes B394, B4 and wild type B131. The biosensor was based on a screen printed carbon electrode (SPE) that was integrated into a flow cell. Enzymes were immobilised on cobalt (II) phthalocyanine (CoPC) modified electrodes by entrapment in a photocrosslinkable polymer (PVA-AWP). The automated flow-based biosensor was successfully used to quantify three organophosphate pesticides (OPs) in milk samples. The OPs used were chlorpyriphos-oxon (CPO), ethyl paraoxon (EPOx) and malaoxon (MOx). The total analysis time for the assay was less than 15 min. Initially, the biosensor performance was tested in phosphate buffer solution (PBS) using B394, B131 and B4 biosensors. The best detection limits were obtained with B394; therefore, this biosensor was used to produce calibration data in milk with three OPs in the concentration range of 5 x 10(-6)M to 5 x 10(-12)M. The limit of detection (LOD) obtained in milk for CPO, EPOx and MOx were 5 x 10(-12)M, 5 x 10(-9)M and 5 x 10(-10)M, respectively, with a correlation coefficient R(2)=0.9910. The automated flow-based biosensor successfully quantified the OPs in different fat-containing milk samples. There were no false positives or false negatives observed for the analytical figures of merit for the constructed biosensors. This method is inexpensive, sensitive, portable, non-invasive and provides real-time results. This analytical system can provide rapid detection of highly toxic OPs in food matrices such as milk.
ESTHER : Mishra_2012_Biosens.Bioelectron_32_56
PubMedSearch : Mishra_2012_Biosens.Bioelectron_32_56
PubMedID: 22221795

Title : Preliminary X-ray analysis of twinned crystals of the Q88Y25_Lacpl esterase from Lactobacillus plantarum WCFS1 - Alvarez_2011_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_67_1436
Author(s) : Alvarez Y , Esteban-Torres M , Acebron I , de Las Rivas B , Munoz R , Martinez-Ripoll M , Mancheno JM
Ref : Acta Crystallographica Sect F Struct Biol Cryst Commun , 67 :1436 , 2011
Abstract : Q88Y25_Lacpl is an esterase produced by the lactic acid bacterium Lactobacillus plantarum WCFS1 that shows amino-acid sequence similarity to carboxylesterases from the hormone-sensitive lipase family, in particular the AFEST esterase from the archaeon Archaeoglobus fulgidus and the hyperthermophilic esterase EstEI isolated from a metagenomic library. N-terminally His(6)-tagged Q88Y25_Lacpl has been overexpressed in Escherichia coli BL21 (DE3) cells, purified and crystallized at 291 K using the hanging-drop vapour-diffusion method. Mass spectrometry was used to determine the purity and homogeneity of the enzyme. Crystals of His(6)-tagged Q88Y25_Lacpl were prepared in a solution containing 2.8 M sodium acetate trihydrate pH 7.0. X-ray diffraction data were collected to 2.24 A resolution on beamline ID29 at the ESRF. The apparent crystal point group was 422; however, initial global analysis of the intensity statistics (data processed with high symmetry in space group I422) and subsequent tests on data processed with low symmetry (space group I4) showed that the crystals were almost perfectly merohedrally twinned. Most probably, the true space group is I4, with unit-cell parameters a = 169.05, b = 169.05, c = 183.62 A.
ESTHER : Alvarez_2011_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_67_1436
PubMedSearch : Alvarez_2011_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_67_1436
PubMedID: 22102251
Gene_locus related to this paper: lacpl-LP.0973

Title : Synthesis of propyl gallate by transesterification of tannic acid in aqueous media catalysed by immobilised derivatives of tannase from Lactobacillus plantarum - Fernandez-Lorente_2011_Food.Chem_128_214
Author(s) : Fernandez-Lorente G , Bolivar JM , Rocha-Martin J , Curiel JA , Munoz R , de Las Rivas B , Carrascosa AV , Guisan JM
Ref : Food Chem , 128 :214 , 2011
Abstract : Immobilised derivatives of tannase from Lactobacillus plantarum were able to catalyse the transesterification of tannic acid by using moderate concentrations of 1-propanol in aqueous media. Transesterification of tannic acid was very similar to transesterification of methyl gallate. The synthetic yield depended on the pH and concentration of 1-propanol, although it did not vary much when using 30% or 50% 1-propanol. Synthetic yields of 45% were obtained with 30% of 1-propanol at pH 5.0. The product was chromatographically pure, and the reaction by-product was 55% pure gallic acid. On the other hand, immobilised tannase was fairly stable under optimal reaction conditions.
ESTHER : Fernandez-Lorente_2011_Food.Chem_128_214
PubMedSearch : Fernandez-Lorente_2011_Food.Chem_128_214
PubMedID: 25214351

Title : Hydrolysis of tannic acid catalyzed by immobilized-stabilized derivatives of Tannase from Lactobacillus plantarum - Curiel_2010_J.Agric.Food.Chem_58_6403
Author(s) : Curiel JA , Betancor L , de Las Rivas B , Munoz R , Guisan JM , Fernandez-Lorente G
Ref : Journal of Agricultural and Food Chemistry , 58 :6403 , 2010
Abstract : A recombinant tannase from Lactobacillus plantarum , overexpressed in Escherichia coli , was purified in a single step by metal chelate affinity chromatography on poorly activated nickel supports. It was possible to obtain 0.9 g of a pure enzyme by using only 20 mL of chromatographic support. The pure enzyme was immobilized and stabilized by multipoint covalent immobilization on highly activated glyoxyl agarose. Derivatives obtained by multipoint and multisubunit immobilization were 500- and 1000-fold more stable than both the soluble enzyme and the one-point-immobilized enzyme in experiments of thermal and cosolvent inactivation, respectively. In addition, up to 70 mg of pure enzyme was immobilized on 1 g of wet support. The hydrolysis of tannic acid was optimized by using the new immobilized tannase derivative. The optimal reaction conditions were 30% diglyme at pH 5.0 and 4 degrees C. Under these conditions, it was possible to obtain 47.5 mM gallic acid from 5 mM tannic acid as substrate. The product was pure as proved by HPLC. On the other hand, the immobilized biocatalyst preserved >95% of its initial activity after 1 month of incubation under the optimal reaction conditions.
ESTHER : Curiel_2010_J.Agric.Food.Chem_58_6403
PubMedSearch : Curiel_2010_J.Agric.Food.Chem_58_6403
PubMedID: 20438129
Gene_locus related to this paper: lacpl-tanL

Title : Production and physicochemical properties of recombinant Lactobacillus plantarum tannase - Curiel_2009_J.Agric.Food.Chem_57_6224
Author(s) : Curiel JA , Rodriguez H , Acebron I , Mancheno JM , de Las Rivas B , Munoz R
Ref : Journal of Agricultural and Food Chemistry , 57 :6224 , 2009
Abstract : Tannase is an enzyme with important biotechnological applications in the food industry. Previous studies have identified the tannase encoding gene in Lactobacillus plantarum and also have reported the description of the purification of recombinant L. plantarum tannase through a protocol involving several chromatographic steps. Here, we describe the high-yield production of pure recombinant tannase (17 mg/L) by a one-step affinity procedure. The purified recombinant tannase exhibits optimal activity at pH 7 and 40 degrees C. Addition of Ca(2+) to the reaction mixture greatly increased tannase activity. The enzymatic activity of tannase was assayed against 18 simple phenolic acid esters. Only esters derived from gallic acid and protocatechuic acid were hydrolyzed. In addition, tannase activity was also assayed against the tannins tannic acid, gallocatechin gallate, and epigallocatechin gallate. Despite L. plantarum tannase representing a novel family of tannases, which shows no significant similarity to tannases from fungal sources, both families of enzymes shared similar substrate specificity range. The physicochemical characteristics exhibited by L. plantarum recombinant tannase make it an adequate alternative to the currently used fungal tannases.
ESTHER : Curiel_2009_J.Agric.Food.Chem_57_6224
PubMedSearch : Curiel_2009_J.Agric.Food.Chem_57_6224
PubMedID: 19601665
Gene_locus related to this paper: lacpl-tanL

Title : Characterization of tannase activity in cell-free extracts of Lactobacillus plantarum CECT 748T - Rodriguez_2008_Int.J.Food.Microbiol_121_92
Author(s) : Rodriguez H , de Las Rivas B , Gomez-Cordoves C , Munoz R
Ref : Int J Food Microbiol , 121 :92 , 2008
Abstract : In foods, tannins are considered nutritionally undesirable. Spectrophotometric methods have been used to detect tannin degradation by L. plantarum strains isolated from food substrates. Enzymatic degradation of tannic acid by L. plantarum CECT 748T was examined in liquid cultures and in cell-free extracts by HPLC. Significative reduction of tannic acid was not observed during incubation in the presence of L. plantarum cells after 7 days incubation. However, tannic acid was effectively degraded by cell-free extracts of L. plantarum during 16 h incubation. We have partially characterized L. plantarum tannase activity by measuring its esterase activity on methyl gallate. Tannase activity was optimal at pH 5.0 and 30 degrees C, and showed nearly 75% of the maximal activity at 50 degrees C. The biochemical characteristics showed by L. plantarum tannase are considered favourable for tannin biodegradation in the food-processing industry.
ESTHER : Rodriguez_2008_Int.J.Food.Microbiol_121_92
PubMedSearch : Rodriguez_2008_Int.J.Food.Microbiol_121_92
PubMedID: 18054106