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Cyclopropane fatty acid synthase from Oenococcus oeni: expression in Lactococcus lactis subsp. cremoris and biochemical characterization.
Arch Microbiol 2015; 197(9):1063-74AM

Abstract

Bacterial cyclopropane fatty acid synthases (CFA synthases) catalyze the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to the double bond of a lipid chain, thereby forming a cyclopropane ring. CFAs contribute to resistance to acidity, dryness, and osmotic imbalance in many bacteria. This work describes the first biochemical characterization of a lactic acid bacterium CFA synthase. We have overexpressed Oenococcus oeni CFA synthase in E. coli in order to purify the enzyme. The optimum cyclopropanation activity was obtained at pH 5.6 and 35.8 °C. The high K(m) (AdoMet) value obtained (2.26 mM) demonstrates the low affinity of O. oeni enzyme toward the L. lactis subsp. cremoris unsaturated phospholipids. These results explain the partial complementation of the L. lactis subsp. cremoris cfa mutant by the O. oeni cfa gene and suggest a probable substrate specificity of the O. oeni enzyme. The current study reveals an essential hypothesis about the specificity of O. oeni CFA synthase which could play a key function in the acid tolerance mechanisms of this enological bacterium.

Authors+Show Affiliations

Unité Mixte de Recherche Procédés Alimentaires et Microbiologiques (UMR A 02.102 AgroSup Dijon-Université de Bourgogne), Equipe Vin Aliment Microbiologie et Stress, Institut Universitaire de la Vigne et du Vin "Jules Guyot", Rue Claude Ladrey, Campus Universitaire Montmuzard, BP 27877, 21078, Dijon, France.Unité Mixte de Recherche Procédés Alimentaires et Microbiologiques (UMR A 02.102 AgroSup Dijon-Université de Bourgogne), Equipe Vin Aliment Microbiologie et Stress, Institut Universitaire de la Vigne et du Vin "Jules Guyot", Rue Claude Ladrey, Campus Universitaire Montmuzard, BP 27877, 21078, Dijon, France. cosette.grandvalet@u-bourgogne.fr. Institut National Supérieur des Sciences Agronomiques de l'Alimentation et de l'Environnement, AgroSup Dijon, Site Epicure, 1 Esplanade Erasme, Campus Universitaire, 21000, Dijon, France. cosette.grandvalet@u-bourgogne.fr.Unité Mixte de Recherche Procédés Alimentaires et Microbiologiques (UMR A 02.102 AgroSup Dijon-Université de Bourgogne), Equipe Vin Aliment Microbiologie et Stress, Institut Universitaire de la Vigne et du Vin "Jules Guyot", Rue Claude Ladrey, Campus Universitaire Montmuzard, BP 27877, 21078, Dijon, France.Unité Mixte de Recherche Procédés Alimentaires et Microbiologiques (UMR A 02.102 AgroSup Dijon-Université de Bourgogne), Equipe Vin Aliment Microbiologie et Stress, Institut Universitaire de la Vigne et du Vin "Jules Guyot", Rue Claude Ladrey, Campus Universitaire Montmuzard, BP 27877, 21078, Dijon, France.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

26294376

Citation

To, Thi Mai Huong, et al. "Cyclopropane Fatty Acid Synthase From Oenococcus Oeni: Expression in Lactococcus Lactis Subsp. Cremoris and Biochemical Characterization." Archives of Microbiology, vol. 197, no. 9, 2015, pp. 1063-74.
To TM, Grandvalet C, Alexandre H, et al. Cyclopropane fatty acid synthase from Oenococcus oeni: expression in Lactococcus lactis subsp. cremoris and biochemical characterization. Arch Microbiol. 2015;197(9):1063-74.
To, T. M., Grandvalet, C., Alexandre, H., & Tourdot-Maréchal, R. (2015). Cyclopropane fatty acid synthase from Oenococcus oeni: expression in Lactococcus lactis subsp. cremoris and biochemical characterization. Archives of Microbiology, 197(9), pp. 1063-74. doi:10.1007/s00203-015-1143-y.
To TM, et al. Cyclopropane Fatty Acid Synthase From Oenococcus Oeni: Expression in Lactococcus Lactis Subsp. Cremoris and Biochemical Characterization. Arch Microbiol. 2015;197(9):1063-74. PubMed PMID: 26294376.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cyclopropane fatty acid synthase from Oenococcus oeni: expression in Lactococcus lactis subsp. cremoris and biochemical characterization. AU - To,Thi Mai Huong, AU - Grandvalet,Cosette, AU - Alexandre,Hervé, AU - Tourdot-Maréchal,Raphaëlle, Y1 - 2015/08/21/ PY - 2015/05/12/received PY - 2015/08/12/accepted PY - 2015/07/09/revised PY - 2015/8/22/entrez PY - 2015/8/22/pubmed PY - 2016/2/5/medline KW - CFA synthase KW - Enzymatic activity KW - Heterologous complementation KW - Oenococcus oeni SP - 1063 EP - 74 JF - Archives of microbiology JO - Arch. Microbiol. VL - 197 IS - 9 N2 - Bacterial cyclopropane fatty acid synthases (CFA synthases) catalyze the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to the double bond of a lipid chain, thereby forming a cyclopropane ring. CFAs contribute to resistance to acidity, dryness, and osmotic imbalance in many bacteria. This work describes the first biochemical characterization of a lactic acid bacterium CFA synthase. We have overexpressed Oenococcus oeni CFA synthase in E. coli in order to purify the enzyme. The optimum cyclopropanation activity was obtained at pH 5.6 and 35.8 °C. The high K(m) (AdoMet) value obtained (2.26 mM) demonstrates the low affinity of O. oeni enzyme toward the L. lactis subsp. cremoris unsaturated phospholipids. These results explain the partial complementation of the L. lactis subsp. cremoris cfa mutant by the O. oeni cfa gene and suggest a probable substrate specificity of the O. oeni enzyme. The current study reveals an essential hypothesis about the specificity of O. oeni CFA synthase which could play a key function in the acid tolerance mechanisms of this enological bacterium. SN - 1432-072X UR - https://www.unboundmedicine.com/medline/citation/26294376/Cyclopropane_fatty_acid_synthase_from_Oenococcus_oeni:_expression_in_Lactococcus_lactis_subsp__cremoris_and_biochemical_characterization_ L2 - https://dx.doi.org/10.1007/s00203-015-1143-y DB - PRIME DP - Unbound Medicine ER -