title: Identification and characterization of carboxyl esterases of gill chamber-associated microbiota in the deep-sea shrimp Rimicaris exoculata by using functional metagenomics.
abstract: The shrimp Rimicaris exoculata dominates the fauna in deep-sea hydrothermal vent sites along the Mid-Atlantic Ridge (depth, 2,320 m). Here, we identified and biochemically characterized three carboxyl esterases from microbial communities inhabiting the R. exoculata gill that were isolated by naive screens of a gill chamber metagenomic library. These proteins exhibit low to moderate identity to known esterase sequences (≤52%) and to each other (11.9 to 63.7%) and appear to have originated from unknown species or from genera of Proteobacteria related to Thiothrix/Leucothrix (MGS-RG1/RG2) and to the Rhodobacteraceae group (MGS-RG3). A library of 131 esters and 31 additional esterase/lipase preparations was used to evaluate the activity profiles of these enzymes. All 3 of these enzymes had greater esterase than lipase activity and exhibited specific activities with ester substrates (≤356 U mg(-1)) in the range of similar enzymes. MGS-RG3 was inhibited by salts and pressure and had a low optimal temperature (30°C), and its substrate profile clustered within a group of low-activity and substrate-restricted marine enzymes. In contrast, MGS-RG1 and MGS-RG2 were most active at 45 to 50°C and were salt activated and barotolerant. They also exhibited wider substrate profiles that were close to those of highly active promiscuous enzymes from a marine hydrothermal vent (MGS-RG2) and from a cold brackish lake (MGS-RG1). The data presented are discussed in the context of promoting the examination of enzyme activities of taxa found in habitats that have been neglected for enzyme prospecting; the enzymes found in these taxa may reflect distinct habitat-specific adaptations and may constitute new sources of rare reaction specificities.
authors.affiliation: Consejo Superior de Investigaciones Científicas (CSIC), Institute of Catalysis, Madrid, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Alcaide
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Anatoli
authors.initials: A
authors.lastname: Tchigvintsev
authors.affiliation: Consejo Superior de Investigaciones Científicas (CSIC), Institute of Catalysis, Madrid, Spain.
authors.forename: Mónica
authors.initials: M
authors.lastname: Martínez-Martínez
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Ana
authors.initials: A
authors.lastname: Popovic
authors.affiliation: Department of Biochemistry, University of Pretoria, Pretoria, South Africa.
authors.forename: Oleg N
authors.initials: ON
authors.lastname: Reva
authors.affiliation: Consejo Superior de Investigaciones Científicas (CSIC), Institute of Catalysis, Madrid, Spain.
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Lafraya
authors.affiliation: Consejo Superior de Investigaciones Científicas (CSIC), Institute of Catalysis, Madrid, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Bargiela
authors.affiliation: Insect Symbiosis Research Group, Max Planck Institute for Chemical Ecology, Jena, Germany.
authors.forename: Taras Y
authors.initials: TY
authors.lastname: Nechitaylo
authors.affiliation: Centro de Investigaciones Biológicas, CSIC, Madrid, Spain.
authors.forename: Ruth
authors.initials: R
authors.lastname: Matesanz
authors.affiliation: Ifremer, Centre de Brest, Laboratoire de Microbiologie des Environnements Extrêmes, REM/DEEP/LM2E, UMR 6197 (Ifremer-CNRS-UBO), ZI de la Pointe du Diable, Plouzané, France.
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: Cambon-Bonavita
authors.affiliation: Université de Bretagne Occidentale, Laboratoire de Microbiologie des Environnements Extrêmes-UMR 6197 (CNRS-Ifremer-UBO), Plouzané, France.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Jebbar
authors.affiliation: Institute for Coastal Marine Environment, CNR, Messina, Italy.
authors.forename: Michail M
authors.initials: MM
authors.lastname: Yakimov
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Alexei
authors.initials: A
authors.lastname: Savchenko
authors.affiliation: School of Biological Sciences, Bangor University, Gwynedd, United Kingdom.
authors.forename: Olga V
authors.initials: OV
authors.lastname: Golyshina
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Alexander F
authors.initials: AF
authors.lastname: Yakunin
authors.affiliation: School of Biological Sciences, Bangor University, Gwynedd, United Kingdom mferrer@icp.csic.es p.golyshin@bangor.ac.uk.
authors.forename: Peter N
authors.initials: PN
authors.lastname: Golyshin
authors.affiliation: Consejo Superior de Investigaciones Científicas (CSIC), Institute of Catalysis, Madrid, Spain mferrer@icp.csic.es p.golyshin@bangor.ac.uk.
authors.forename: Manuel
authors.initials: M
authors.lastname: Ferrer
mesh_terms: D000818:Animals; D001267:Atlantic Ocean; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D033364:Decapoda / Q000382:microbiology*; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D005880:Gills / Q000382:microbiology*; D060086:Hydrothermal Vents; D054892:Metagenome; D056186:Metagenomics; D064307:Microbiota; D008969:Molecular Sequence Data; D012492:Salts / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D012492:Salts; D002265:Carboxylic Ester Hydrolases