DE-AC02-05CH11231, Lawrence Livermore National Laboratory under Contract No. DE-AC52-07NA27344, and Los Alamos National Laboratory under contract No. DE-AC02-06NA25396, UT-Battelle and Oak Ridge National Laboratory under contract DE-AC05-00OR22725, as well as German Research Foundation (DFG) INST 599/1-2.
E. lignolyticus�� http://www.selleckchem.com/products/dorsomorphin-2hcl.html SCF1 was isolated from soil collected from the Short Cloud Forest site in the El Yunque experimental forest, part of the Luquillo Long-Term Ecological Research Station in Luquillo, Puerto Rico, USA (Table 1). Soils were diluted in water and inoculated into roll tubes containing MOD-CCMA media with alkali lignin as the source of carbon. MOD-CCMA media consists of 2.8 g L-1 NaCl, 0.1 g L-1 KCl, 27 mM MgCl2, 1 mM CaCl2, 1.25 mM NH4Cl, 9.76 g L-1 MES, 1.1 ml L-1 K2HPO4, 12.

5 ml L-1 trace minerals [19,20], and 1 ml L-1 Thauer��s vitamins [21]. Tubes were incubated at room temperature for up to 12 weeks, at which point the colony was picked, grown in 10% tryptic soy broth (TSB), and characterized. Table 1 Classification and general features of ��Enterobacter lignolyticus�� SCF1 When grown on 10% TSB agar plates, SCF1 colonies are translucent white, slightly irregular in shape with wavy margins, and have a shiny smooth surface. SCF1 was determined to be a non-sporulating strain based on a Pasteurization test. To do this, a suspension of SCF1 cells was heated at 80��C for 10 minutes. 5��l of heated culture and non-heated control culture were both spotted onto 10% TSB agar and incubated for growth for 3 days at room temperature.

The non-heated cells grew while the heated culture did not, indicating the absence of heat-resistant spores. For initial genotyping and for validating the isolation, the small subunit ribosomal RNA gene was sequenced by Sanger sequencing using the universal primers 8F and 1492R [22].The 16S rRNA sequence places ��Enterobacter Carfilzomib lignolyticus�� SCF1 in the family Enterobacteriaceae. However, 16S rRNA sequence is not sufficient to clearly define the evolutionary history of this region of the Gammaproteobacteria, and initially led to the incorrect classification of ��E. lignolyticus�� SCF1 as a member of the Enterobacter cloacae species. We have rectified its phylogenetic placement using the MicrobesOnline species tree [23], which is generated using 69 single-copy near-universal protein families [24] aligned by MUSCLE [25] with tree construction using FastTree-2 [26] (Figure 1). Figure 1 Phylogenetic tree highlighting the position of ��Enterobacter lignolyticus�� SCF1 relative to other type and non-type strains within the Enterobacteriaceae. Strains shown are those within the Enterobacteriaceae having corresponding NCBI …