Draft genome sequence of cellulolytic and xylanolytic Paenibacillus sp. A59, isolated from a decaying forest soil from Patagonia, Argentina

SILVINA GHIO; ALFREDO MARTINEZ CÁCERES; PAOLA TALIA; DANIEL GRASSO; ELEONORA CAMPOS

Genome Announcement

University of Illinois

Lugar: Chicago; Año: 2015 vol. 3 p. 1 - 2

Microbial cellulases have become important for many applicationsin food processing, animal feed, pulp and paper industries,and second generation biofuels production (1). In particular,bacteria are promising sources of enzymes for biomass bioconversion.Membersof the Paenibacillus genus have been isolatedfrom soiland plant related sources (2) and have been reported to producemany extracellular enzymes for industrial applications (3).In our study, a novel cellulolytic and xylanolytic strain,named Paenibacillus sp. A59, was isolated from a previouslycharacterized cellulolytic bacterial consortium, obtained fromdecaying forest soil in the Patagonia region, Argentina (4).These bacteria are Gram-positive endospore-forming, facultativeanaerobe bacillus. Based on 16S rRNA gene sequence analysis,it formed a cluster with P. taichungensis (gi|343199050|)and P. pabuli (gi|343200166|). A crude enzymatic extract ofPaenibacillus sp. A59 was capable of converting pre-treated sugarcaneagricultural residue to simple sugars such as xylobiose, xylose,cellobiose, and arabinose, demonstrating its potential forlignocellulosic biomass deconstruction (S. Ghio et al., unpublishedresults).Bacterial genomic DNA was obtained from a 24 h culture inLuria Bertoni broth, by a commercial extraction kit (WizardGenomic DNA Extraction kit, Promega). Genome sequencingwas performed using Illumina MiSeq platform. A total of4,201,470 paired-end reads were generated, with an average lengthof 250 bp, achieving 110-fold coverage of the genome. The rawreads were trimmed using Trimmomatic version 0.33 (5) and assembledde novo using Velvet version 1.2.10 (6), producing 83contigs with an accumulated length of 7,087,589 (N50 317,284 bp)and an average GC content of 46%.The functional annotation was performed with the Rapid Annotationsusing Subsystems Technology (RAST) server, version 2.0 (7) andNCBI Prokaryotic Genome Annotation Pipeline (http://www.ncbi.nlm.nih.gov/genome/annotation_prok/), resulting in 6,424predicted coding sequences, 39% of which were assigned to 447 subsystems.Also, 48 RNA sequences were identified: 46 tRNAs and onecopy of 23S/5S and 16S rRNA genes. A comparison of Paenibacillussp. A59 genome sequence with others available in the RAST serverresulted in Geobacillus sp. Y412MC10 (score 524) and Paenibacillussp. oral taxon 786 strain D14 (score 461) as closest neighbors. Severalgenes codingfor potential glycoside hydrolases (GHs), as classified bythe CAZY database (8), were found. Among them, 4 endoxylanases(GH10:3, GH11:1), 2 endoglucanases (GH5:1, GH9:1), 2 cellobiohydrolases(GH6:1, GH48:1), 13 -xylosidases/arabinofuranosidases(GH43:12, GH52:1), and 7 -glucosidases (GH1:1, GH3:6)were the most relevant enzymes identified for lignocellulosic biomassdeconstruction. These preliminary results correlate with theobserved enzymatic activity of Paenibacillus sp. A59 and highlightthe role of Paenibacillus genus as a source of (hemi) cellulolyticenzymes for biotechnological and industrial applications.Nucleotide sequence accession numbers. The draft genomesequence of Paenibacillus sp. A59 has been deposited in DDBJ/EMBL/GenBank under the accession no. LITU00000000. The versiondescribed in this paper is LITU01000000.A