Genome Sequence of Anaerobacillus macyae JMM-4T (DSM 16346), the First Genomic Information of the Newly Established Genus Anaerobacillus.

Anaerobacillus macyae JMM-4(T) (DSM 16346) is a Gram-positive, spore-forming, strictly anaerobic, and arsenate-respiring bacterium. Here, we report the 4.26-Mb genome sequence of A. macyae JMM-4(T), which is the first genome information of the newly established genus Anaerobacillus.

GENOME ANNOUNCEMENT

The Bacillus-like strain JMM-4T was isolated from an arsenic-contaminated environment in Bendigo, Victoria, Australia (1), and was formerly identified as one member of the genus Bacillus, for which the name Bacillus macyae sp. nov. was proposed (2). In 2009, the new genus Anaerobacillus within the family Bacillaceae was established (3). Meanwhile, B. macyae along with Bacillus alkalidiazotrophicus (4) and Bacillus arseniciselenatis (5) were transferred to the new genus Anaerobacillus as A. macyae comb. nov., A. alkalidiazotrophicus comb. nov., and A. arseniciselenatis comb. nov., respectively (3).

Counting A. alkalilacustre, four species were assigned taxonomically to this genus. The mainly common features shared by all the four species of the genus Anaerobacillus are strictly anaerobic or aerotolerant, halotolerant or moderately halophilic, obligate or moderately alkaliphilic, chemo-organotrophic, and diazotrophic (3). More importantly, these species are capable of anaerobic respiration with arsenate as a terminal electron acceptor, using a variety of substrates as the electron donors (2, 3). Given the taxonomic history, physiological properties, and no available genomic information of A. macyae, its type strain F12T was selected as one of the research objects in our “genome sequencing project for genomic taxonomy and phylogenomics of Bacillus-like bacteria.” Here, we present the high-quality draft genome sequence of A. macyae JMM-4T (DSM 16346).

The genome sequencing of A. macyae JMM-4T (DSM 16346) was performed via the Illumina Hiseq 2500 system. A PCR free-DNA library with insert sizes of 500 bp was constructed and sequenced using the 2 × 150-bp paired-end sequencing strategy. After filtering of the 567.42 Mb of raw data, 567.25 Mb of clean data were obtained, providing more than 100-fold coverage. The reads were assembled via the SOAPdenovo software version 1.05 (6), using a key parameter K setting at 77. Through the data assembly, 16 scaffolds with a total of 4,261,692 bp were obtained, and the scaffold N50 was 1,507,096 bp. The average length of the scaffolds was 266,355 bp, and the longest and shortest scaffolds were 2,061,986 bp and 1,383 bp, respectively. Moreover, 91.78% clean reads were aligned back to the genome, which covered 99.99% of the sequence.

Genome annotation was completed using the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (PGAAP). The tRNA and rRNA genes were identified by tRNAscan-SE (7) and RNAmmer (8), respectively. A total of 4,208 genes were predicted, including 3,973 coding sequences (CDS), 145 pseudogenes, 81 tRNAs, and 8 rRNA genes. The average DNA G+C content was 39.85%, with a slight difference from the value of 37 mol% acquired by HPLC determination (2).

The genome sequence of A. macyae JMM-4T (DSM 16346) provides useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria. Moreover, the genomic information will promote research on the biological significance and application in environmental biotechnology of the arsenate-respiring activity of the Anaerobacillus species.

Nucleotide sequence accession numbers.

This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number LELK00000000. The version described in this paper is version LELK01000000.