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Covalently Linked Polyamines in the Cell Wall Peptidoglycan of the Anaerobes Belonging to the Order Selenomonadales

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Covalently Linked Polyamines in the Cell Wall Peptidoglycan of the Anaerobes Belonging to the Order Selenomonadales J. Gen. Appl. Microbiol., 58, 339‒347 (2012) Short Communication Covalently linked polyamines in the cell wall peptidoglycan of the anaerobes belonging to the order Selenomonadales Koei Hamana,1,* Takashi Itoh,2 Mitsuo Sakamoto,2 and Hidenori Hayashi 1 1 Faculty of Engineering, Maebashi Institute of Technology, Maebashi, Gunma 371‒0816, Japan 2 Japan Collection of Microorganisms, RIKEN BioResource Center (BRC), Wako, Saitama 351‒0198, Japan (Received May 1, 2012; Accepted May 20, 2012) Key Words—Acidaminococcaceae; peptidoglycan; polyamine; Selenomonadales; Veillonellaceae In our studies on cellular polyamine analysis in bac- outer membrane of the envelope in typical Gram-neg- teria (the domain Bacteria) as a source of chemotaxo- ative taxa (Takatsuka and Kamio, 2004; Kamio et al., nomic information (Hamana, 1999a, 2002a; Hamana 2007). On the other hand, intracellular polyamine and Hosoya, 2006; Hamana and Matsuzaki, 1992; Ha- pools in these Gram-negative anaerobes were not re- mana et al., 2006; Hosoya et al., 2006), covalently fl ected in their covalently linked polyamine levels (Ha- linked putrescine, cadaverine, spermidine and/or ag- mana, 1999b; Hamana et al., 2002). The relation be- matine in cell wall peptidoglycan have been found in tween acid-extractable ionic-binding polyamines and Gram-negative anaerobes belonging to the 13 genera covalently linked polyamine components should be Acetonema, Acidaminococcus, Anaeromusa, Anaerovi- presented for the evaluation of bacterial polyamine brio, Dialister, Dendrosporobacter, Megasphaera, distribution profi les as a chemotaxonomic marker. To Pectinatus, Propionispira, Selenomonas, Sporomusa, obtain additional distribution catalogues of covalently Veillonella and Zymophilus (Hamana, 1999b; Hamana linked polyamines within the order Selenomonadales et al., 2002). It has been revealed that one of these consisting of the two families Acidaminococcaceae polyamines binds covalently to the α-carboxy group of and Veillonellaceae (NCBI website, 2012), polyamines the D-glutamic acid residue of a peptidoglycan in Sele- of 17 new species including the fi ve newly validated nomonas ruminantium, Veillonella parvula and An- genera Desulfosporomusa, Megamonas, Mitsuokella, aerovibrio lipolytica (Hirao et al., 2000; Kamio, 1987; Propionispora and Sporolituus in the order Selenom- Kamio and Nakamura, 1987; Kamio et al., 1981; Taka- onadales were analyzed. Polyamines in the acid-ex- tsuka and Kamio, 2004). These Gram-negative anaer- tracts from whole cells and in the acid-hydolysates of obes belonging to the order Selenomonadales (for- cell residues after the acid extraction, were determined merly the Sporomusa subbranch), phylogenetically after growth at different temperatures and in culture located in the phylum Firmicutes, a Gram-positive tax- media supplemented with diaminopropane, diamino- on (NCBI website, 2012), however, lack murein lipo- hexane, spermidine, spermine or agmatine. Seven protein which maintains the structural integrity of the Gram-positive anaerobes belonging to the order Clostridiales of the phylum Firmicutes were used as a related anaerobic bacterium. * Address reprint requests to: Dr. Koei Hamana, Faculty of Engineering, Maebashi Institute of Technology, Maebashi, Gun- The organisms were provided by the RIKEN BRC ma 371‒0816, Japan. through the National Bio-Resource Project of the E-mail: [email protected] MEXT, Japan and grown anaerobically at 25‒60°C in 340 HAMANA et al. Vol. 58 Fig. 1. HPLC analysis of the PCA extract (PCA) and the HCl-hydrolysate of the cell residue after the PCA extraction (HCl-Res) of Selenomonas fl ueggei JCM 8544 (1) and Veillonella ratti JCM 6512 (2) grown at 37°C in GAM medium in the absence (control) (A), presence of 5 mM agmatine (B), 5 mM diaminopropane (C) or 5 mM diaminohexane (D). Abbreviations for polyamines are shown in Table 1. GAM Broth (GAM) (Nissui Pharmaceutical Co., Tokyo, website, 2012), using Anaerobic Jar KJ-1 (Tomy Seiko, Japan), Modifi ed GAM Broth (MGAM) (Nissui), Tokyo) under H2 gas. Cells in the stationary phase polyamine-free /arginine-rich synthetic RPMI 1640 me- were harvested from a 50‒100 ml liquid culture by cen- dium (Nissui) (Hamana et al., 2007) supplemented trifugation at 3,000 × g and washed fi ve times with an with 0.03% L-cysteine and 0.03% sodium thioglycol- isotonic saline or PBS (Nissui Pharmaceutical Co.). late (1640-Cys-TG) and JCM media (listed on the JCM The cell pellets were homogenized in equal volumes 2012 Table 1. Acid-extractable polyamines from cells and polyamines released by acid-hydrolysis from cell residue after acid-extraction in the anaerobes belonging to the orders Selenomonadales and Clostridiales of the phylum Firmicutes, and covalently linked polyamines. Temp. Fraction Polyamines (μmol/g wet weight) Covalently linked Organism Med. (°C) (Ref.) Put Cad Spd Spm Agm Dap Dah polyamines Order Selenomonadales Family Acidaminococcaceae Acidaminococcus fermentans ATCC 25085T GAM 37 (a) Put, Cad, Spd Family Veillonellaceae Acetonema longum ATCC 51454T ATCC1249 37 (a) Cad Anaerovibrio burkinobensis ATCC 51455T GAM 37 (a) Put Anaerovibrio glycerini ATCC 51177T GAM 37 (a) Put, Cad Anaerovibrio lypolyticus ATCC 33276T GAM 37 (b) Cad, Spd (Anaerovibrio lypolytica) Anaeromusa acidaminophila ATCC 43704T GAM 37 (b) Put, Cad, Spd (Selenomonas acidaminophila) in Polyamines Dendrosporobacter quercicolus ATCC 25974T GAM 37 (b) Cad (Clostridium quercicolus) Desulfosporomusa polytropa JCM 12564T MGAM 25 PCA 0.00 0.04 0.12 0.00 0.00 0.00 0.00 Put, Cad, Spd HCl-Res 0.35 0.85 0.30 0.00 0.00 0.00 0.00 Dialister pneumosintes JCM 10004T MGAM 30 (a) Put, Cad Selenomonadales Dialister succinatiphilus JCM 15077T MGAM 25 PCA 0.90 0.03 0.05 0.00 0.00 0.00 0.00 Put HCl-Res 0.25 0.00 0.00 0.00 0.00 0.00 0.00 Dialister invisus JCM 17566T JCM282 30 PCA 0.05 0.54 0.00 0.00 0.00 0.00 0.00 Cad HCl-Res 0.00 0.15 0.00 0.00 0.00 0.00 0.00 Dialister microaerophilus JCM 17567T JCM282 30 PCA 0.05 0.45 0.00 0.07 0.05 0.00 0.00 Cad HCl-Res 0.00 0.15 0.00 0.00 0.00 0.00 0.00 Dialister propionicifaciens JCM 17568T JCM282 30 PCA 0.02 0.33 0.00 0.00 0.00 0.00 0.00 Cad HCl-Res 0.00 0.10 0.00 0.00 0.00 0.00 0.00 Megamonas funiformis JCM 14723T MGAM 25 PCA 0.52 0.53 0.65 0.00 0.05 0.00 0.00 Put, Cad HCl-Res 0.17 0.85 0.02 0.00 0.06 0.00 0.00 Megamonas hypermegale ATCC 25560T GAM 37 PCA 0.00 0.40 0.05 0.00 0.00 0.00 0.00 Cad (Bacteroides hypermegas) HCl-Res 0.00 0.84 0.00 0.00 0.00 0.00 0.00 Megasphaera cerevisiae JCM 6130T GAM 30 (a) Put, Cad Megasphaera cerevisiae JCM 6129 GAM 30 (a) Put, Cad Megasphaera elsdenii JCM 1772T GAM 30 (a) Put, Cad Mitsuokella multacida JCM 2054T MGAM 25 PCA 0.60 0.45 0.85 0.01 0.05 0.00 0.00 Put, Cad (Bacteroides multiacidus) HCl-Res 0.26 0.74 0.03 0.00 0.05 0.00 0.00 Mitsuokella jalaludinii JCM 12226T MGAM 25 PCA 0.60 0.45 0.85 0.00 0.05 0.00 0.00 Put, Cad HCl-Res 0.26 0.74 0.03 0.00 0.05 0.00 0.00 Pectinatus frisingensis ATCC 33332T GAM 30 (a) Cad Pectinatus cerevissiiphilus NCIMB12297T GAM 30 (a) Put, Cad 341 342 Table 1(Continued) Temp. Fraction Polyamines (μmol/g wet weight) Covalently linked Organism Med. (°C) (Ref.) Put Cad Spd Spm Agm Dap Dah polyamines Propionispira arboris ATCC 33732T GAM 30 (a) Put, Cad, Spd Propionispora vibrioides JCM 17185T JCM841 30 PCA 0.00 0.03 0.20 0.00 0.00 0.00 0.00 Put, Cad HCl-Res 0.15 0.85 0.01 0.00 0.00 0.00 0.00 Selenomonas artemidis JCM 8543T GAM 30 (b) Cad Selenomonas bovis JCM 15470T MGAM 25 PCA 0.00 0.05 0.20 0.00 0.06 0.00 0.00 Cad HCl-Res 0.02 1.40 0.00 0.00 0.00 0.00 0.00 Selenomonas dianae JCM 8542T GAM 30 (b) Cad, Spd Selenomonas fl ueggei JCM 8544T GAM 30 PCA (b) 0.05 0.07 0.76 0.00 0.01 0.00 0.00 Put, Cad, Spd, Agm HCl-Res(b) 0.53 0.15 0.70 0.00 0.36 0.00 0.00 GAM 25 PCA 0.03 0.02 0.14 0.00 0.30 0.00 0.00 Put, Cad, Spd, Agm HCl-Res 0.08 0.07 0.16 0.00 0.78 0.00 0.00 GAM (control) 37 PCA 0.01 0.02 0.25 0.00 0.42 0.00 0.00 Spd, Agm HCl-Res 0.02 0.02 0.20 0.00 0.70 0.00 0.00 GAM+5 mM Agm 37 PCA 0.41 0.02 0.22 0.00 0.42 0.00 0.00 Put, Cad, Spd, Agm HCl-Res 0.49 0.14 0.25 0.00 0.35 0.00 0.00 H GAM+5 mM Dap 37 PCA 0.20 0.00 0.51 0.00 0.30 0.27 0.00 Spd, Agm, Dap AMANA HCl-Res 0.01 0.00 0.10 0.00 0.58 0.06 0.00 GAM+5 mM Dah 37 PCA 0.00 0.00 0.20 0.00 0.00 0.00 0.67 Spd, Agm, Dah etal. HCl-Res 0.00 0.02 0.05 0.00 0.10 0.00 1.02 GAM (control) 25 PCA 0.28 0.01 0.80 0.01 0.04 0.00 0.00 Put, Spd, Agm HCl-Res 0.35 0.01 0.13 0.00 0.34 0.00 0.00 GAM+5 mM Agm 25 PCA 0.30 0.00 0.35 0.00 0.25 0.00 0.00 Put, Spd, Agm HCl-Res 0.70 0.04 0.10 0.00 0.60 0.00 0.00 GAM+5 mM Dap 25 PCA 0.05 0.01 0.55 0.00 0.03 0.17 0.00 Put, Spd, Agm, Dap HCl-Res 0.18 0.02 0.15 0.00 0.38 0.10 0.00 GAM+5 mM Dah 25 PCA 0.00 0.00 0.13 0.00 0.03 0.00 0.60 Spd, Agm, Dah HCl-Res 0.00 0.00 0.10 0.00 0.20 0.00 0.61 Selenomonas infelix JCM 8545T GAM 30 (b) Put, Cad, Spd Selenomonas noxia JCM 8546T GAM 30 (b) Put, Cad Selenomonas ruminantium JCM 6582T GAM 30 PCA (b) 0.35 1.45 1.00 0.00 0.00 0.00 0.00 Put, Cad HCl-Res (b) 0.26 1.35 0.03 0.00 0.00 0.00 0.00 GAM 25 PCA 0.00 1.52 0.07 0.00 0.02 0.00 0.00 Cad HCl-Res 0.00 0.12 0.00 0.00 0.00 0.00 0.00 GAM (control) 37 PCA 0.10 0.85 0.02 0.00 0.02 0.00 0.00 Cad HCl-Res 0.00 0.15 0.00 0.00 0.00 0.00 0.00 GAM+5 mM Spd 37 PCA 0.30 0.10 0.50 0.00 0.00 0.00 0.00 Cad, Spd HCl-Res 0.03 0.10 0.06 0.00 0.04 0.00 0.00 58 Vol.
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