Agrococcus jenensis gen. nov., sp. nov., a New Genus of Actinomycetes with Diaminobutyric Acid in the Cell Wall

Abstract

Actinomycetes are considered an inexhaustible source of chemically diverse secondary metabolites. In addition to the genera Streptomyces, Micromonospora, and Actinoplanes, coryneform and nocardioform bacteria have been used in screening programs for new compounds of medical and biotechnological importance (5, 8).In the course of a program aimed at isolating actinomycetes that have potential for producing novel bioactive compounds, we isolated a large number of strains from different soils. In about 65 of these strains diaminobutyric acid (DAB) was the diagnostic diamino acid in the peptidoglycan. These strains had morphological and chemotaxonomic characteristics that placed them near the genera Agromyces and Clavibacter.In this paper we describe the isolation and characterization of two strains which differed markedly from the members of the genera Agromyces (27), Clavibacter, and Rathayibacter (26) that have been described. On the basis of our morphological, physiological, and biochemical data, as well as the results of our 16s ribosomal DNA (rDNA) analysis, we concluded that these strains belong to a new genus and species, for which we propose the name Agrococcus jenensis. These strains have been deposited in the German Collection of Microorganisms and MATERIALS AND METHODSBacterial strains and cultural conditions. Strain 2002-39/lT was isolated from a sample of frozen compost soil obtained near Jena, Germany, at a depth of about 10 cm. Isolation of this organism involved dilution plating on nutrient agar containing 2% peptone, a pancreatic digest (meat, fish), 0.5% NaCI, and 1.2% agar. Strain ST54 was isolated from the sandstone surface of the Alte Pinakothek building in Munich, Germany. The Agromyces and Cluvibacter type strains which we used in this study are listed in Table 1. General laboratory cultivation was performed on solid medium or in liquid rich (R) medium (25) containing 1% Bacto Peptone (Difco Laboratories), 0.5% yeast extract, 0.5% Casamino Acids, 0.2% meat extract, 0.5% malt extract, 0.2% glycerol, 0.1% MgSO, -7H,O, and 0.005% Tween 80 (pH 7.2) at 28°C.Morphological and physiological characteristics. Cell morphology was determined by examining cultures of different ages by phase-contrast microscopy. Colony morphology was studied by using a stereomicroscope. For scanning electron microscopy an 18-h-old culture of strain 2002-39/lT on an agar plate was suspended in a phosphate-buffered salt solution. The cells were fixed with 0 5 % glutaraldehyde, washed, and dehydrated in a series containing increasing concentrations of ethanol. After sputter coating with gold-palladium, the cells were observed with a Zeiss model 962 scanning electron microscope.

Retymicin, Galtamycin B, Saquayamycin Z and Ribofuranosyllumichrome, Novel Secondary Metabolites from Micromonospora sp. Tü 6368

“…Diaminopimelic acid [11] and the composition of cell-wall sugars [12] was identified by TLC of the whole-organism hydrolysate. Fatty acids were determined by GC-MS [13,14], and menaquinones by reversed-phase HPLC [15,16]. 16S rRNA gene amplification and determination of the almost complete sequence (1495 bp) were carried out according to Rainey et al (1996) [17].…”

Rhodococcus defluvii sp. nov., isolated from wastewater of a bioreactor and formal proposal to reclassify [Corynebacterium hoagii] and Rhodococcus equi as Rhodococcus hoagii comb. nov.

“…Menaquinones were extracted as described by Collins et al (1977) and analysed by HPLC (Groth et al, 1996). The isolate contained dihydrogenated menaquinones with eight isoprene units [MK-8(H 2 )] as the major isoprenologue and minor amounts of MK-6(H 2 ) and MK-7(H 2 ).…”

“…2). Determination of the isomers of diaminopimelic acid and sugars in whole-organism hydrolysates were performed as described by Groth et al (1996). Detection of mycolic acids was performed according to the method of Minnikin et al (1975) with some modifications by thin layer chromatography of their methanolic esters.…”

Virgibacillus kekensis sp. nov., a moderately halophilic bacterium isolated from a salt lake in China

“…Polar lipids were extracted according to the method of Minnikin et al (1979), and were identified by two-dimensional TLC and spraying with specific reagents (Collins & Jones, 1980). Menaquinones were analysed by HPLC as described by Groth et al (1996), and the menaquinones of V. marismortui DSM 12325 T and V. halodenitrificans DSM 10037 T were analysed in parallel with those of strain YIM kkny16 T under identical conditions. Cells were prepared as described by Lee et al (2006) and An et al (2007), and cellular fatty acid methyl esters were identified according to the standard protocol of the Microbial Identification system (MIDI; Microbial ID).…”