De novo assembly and comparative genome analysis for polyhydroxyalkanoates-producing Bacillus sp. BNPI-92 strain.

BACKGROUND: Certain Bacillus species play a vital role in polyhydroxyalkanoate (PHA) production. However, most of these isolates did not properly identify to species level when scientifically had been reported. RESULTS: From NGS analysis, 5719 genes were predicted in the de novo genome assembly. Based on genome annotation using RAST server, 5,527,513 bp sequences were predicted with 5679 bp number of protein-coding sequence. Its genome sequence contains 35.1% and 156 GC content and contigs, respectively. In RAST server analysis, subsystem (43%) and non-subsystem coverage (57%) were generated. Ortho Venn comparative genome analysis indicated that Bacillus sp. BNPI-92 shared 2930 gene cluster (core gene) with B. cereus ATCC 14579 T (AE016877), B. paranthracis Mn5T (MACE01000012), B. thuringiensis ATCC 10792 T (ACNF01000156), and B. antrics Amen T (AE016879) strains. For our strain, the maximum gene cluster (190) was shared with B. cereus ATCC 14579 T (AE016877). For Ortho Venn pair wise analysis, the maximum overlapping gene clusters thresholds have been detected between Bacillus s p.BNPI-92 and Ba. cereus ATCC 14579 T (5414). Average nucleotide identity (ANI) such as OriginalANI and OrthoANI, in silicon digital DND-DNA hybridization (isDDH), Type (Strain) Genome Server (TYGS), and Genome-Genome Distance Calculator (GGDC) were more essentially related Bacillus sp. BNPI-92 with B. cereus ATCC 14579 T strain. Therefore, based on the combination of RAST annotation, OrthoVenn server, ANI and isDDH result Bacillus sp.BNPI-92 strain was strongly confirmed to be a B. cereus type strain. It was designated as B. cereus BNPI-92 strain. In B. cereus BNPI-92 strain whole genome sequence, PHA biosynthesis encoding genes such as phaP, phaQ, phaR (PHA synthesis repressor phaR gene sequence), phaB/phbB, and phaC were predicted on the same operon. These gene clusters were designated as phaPQRBC. However, phaA was located on other operons. CONCLUSIONS: This newly obtained isolate was found to be new a strain based on comparative genomic analysis and it was also observed as a potential candidate for PHA biosynthesis.

Draft genome sequence and potential identification of a biosurfactant from Brevibacterium casei strain LS14 an isolate from fresh water Loktak Lake.

This study reports the whole-genome sequencing and sequence analysis of a bacterial isolate Brevibacterium casei strain LS14, isolated from Loktak Lake, Imphal, India. The de novo assembled genome reported in this paper featured a size of 3,809,532 bp, has GC content of 68% and contains 3602 genomic features, including 3551 protein-coding genes, 46 tRNA and 5rRNA. A biosurfactant biosynthesis gene cluster in the genome of the isolated strain was identified using AntiSMASH online tool V3.0.5 and KAAS (KEGG Automatic Annotation Server). The presence of biosurfactant was demonstrated by drop collapse, oil displacement and emulsification index. Subsequent chemical characterization using FTIR and LC-MS analyses revealed surfactin and terpene containing biosurfactant moieties. Also, the presence of genes involved in terpenoid synthesis pathway in the genome sequence may account for biosurfactant terpenoid backbone, but genes for later-stage conversion of terpenoid to biosurfactant were not ascertained. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02867-9.

Draft Genome Sequence of the Plant Growth-Promoting Rhizobacterium Klebsiella sp. Strain KBG6.2, Imparting Salt Tolerance to Rice.

Klebsiella sp. strain KBG6.2 is a potential salt-tolerant, plant growth-promoting rhizobacterium isolated from a rice field in Konark, Odisha, India. Here, we report the whole-genome sequencing of Klebsiella sp. strain KBG6.2, which has a 5.038-Mb genome containing 4,867 predicted protein-coding sequences and 79 RNA genes.

Comparative Genomics of a Paddy Field Bacterial Isolate Ochrobactrum sp. CPD-03: Analysis of Chlorpyrifos Degradation Potential.

Ochrobactrum genus is known to catabolize aromatic compounds. This study reports a complete genome sequence of Ochrobactrum sp. CPD-03 (~ 4.6 Mb of chromosomal features) responsible for chlorpyrifos (CP) isolated form a paddy field (20.3588° N, 85.8333° E) in Bhubaneswar, India. A comparative genomics approach was performed between CPD-03 and eight closely related genomes of other Ochrobactrum strains in order to deepen our knowledge, to establish its phylogenetic and functional relationships. The involvement of CP degrading genes indicated a versatile role of CPD-03 in additional field trails. This research would provide the genetic information for its use in natural environment for the depletion of organophosphorus (OP) compounds.

Rheinheimera pleomorphica sp. nov., a Novel Alkali-Tolerant Bacteria Isolated from Chilika Lake, India.

A novel Gram-negative gamma-proteobacterium, non-sporulating motile, rod or coccus-shaped bacterium designated as strain PKS7T was isolated from a sediment sample collected from Chilika Lake, Odisha, India and characterized taxonomically using a polyphasic approach. The major quinone was Q8 and major cellular fatty acids were C16:0, C17:0, C15:1w8c, C17:1w8c, C12:03-OH. The chemotaxonomic features confirmed the isolate to be a member of genus Rheinheimera. 16SrRNA gene sequence of strain PKS7T was closest in similarity to R. aquimaris SW-353T (99.36% identity), R. muenzenbergensis E49T (98.63%), R. nanhaiensis E407-8T (98.35%), R. japonica KMM 9513T (98.35%) and R. baltica DSM-14885T (98.08%). The 16S rRNA gene sequence-based phylogenetic analysis and sequence similarity between the isolated strain and type strains also revealed its affiliation to genus Rheinheimera. DNA-DNA relatedness with closest type strain R. aquimaris SW-353T was 25.0% (±3.40) and in silico DDH showed values in the range of 17.7-37.1% with the type strains of the genus Rheinheimera for which whole genome sequence are available. Strain PKS7T was also distinguished by a multi-locus sequence analysis (MLST) by alingning gyrB gene sequences of the closest type strains of Rheinheimera. The draft genome of strain PKS7T contained 32 contigs of total size 3,963,569 bp comprising of 3763 predicted coding sequences with a G + C content of 50.7 mol%. Comparision of phenotypic and genotypic data with its closest neighbours and closely related species confirm the strain PKS7T to be recognised as a novel species within the genus Rheinheimera, for which the name Rheinheimera pleomorphica sp. nov. is proposed. The type strain is PKS7T (= KCTC 42365 = JCM 30460).

An investigation for recovery of polyhydroxyalkanoates (PHA) from Bacillus sp. BPPI-14 and Bacillus sp. BPPI-19 isolated from plastic waste landfill.

Bio-plastic synthesis from renewable and cheap agro-based materials is a sustainable solution for replacing conventionally produced plastic with environmental contamination. The current study was aimed at screening and characterization of Polyhydroxyalkanoates (PHA) producing bacterial isolates, evaluation of their potential and recovery of PHA using the isolates. The PHA compounds were characterized using FT-IR. Based on 16SrRNA sequence analyses the isolates were designated as Bacillus sp. BPPI-14 and Bacillus sp. BPPI-19. The isolates were gram-positive, rod-shaped, endospore former, and citrate test positive. Intracellular PHA granules were observed when these isolates were stained with Sudan black B (SBB) and Nile blue A (NBA) preliminary and specific staining dyes, respectively. Effect of pH, temperature and carbon sources on the PHA production by the isolates BPPI-14 and BPPI-19 was studied. Maximum PHA production was recorded for Glucose (49.46±2.79%) by Bacillus sp. BPPI-14 and followed by molasses (45.86±2.17%) by Bacillus sp. BPPI-19, respectively at 37°C and pH7. The obtained PHA polymers were confirmed by preparation of plastic films for both the isolates. Fourier transform infrared spectrum for BPPI-14 and BPPI-19 showed the peak (carboxylic acid group) at 1706-1719.39cm-1 was a characteristic feature of PHA and corresponds functional group (C=O).

Taxonomic description and genome sequence of Halobacillus marinus sp. nov., a novel strain isolated from Chilika Lake, India.

moderately halophilic spore forming, motile, Gram-positive, rod-shaped bacterial strain designated as KGW1T was isolated from water sample of Chilika Lake and characterized taxonomically using polyphasic approach. The strain grew in the presence of 0-25% (w/v) NaCl in marine salt agar media, hydrolyzes casein, and gelatin and shows presence of alkaline proteases. The major cell wall menaquinone was MK7 and major cellular fatty acids were anteiso-C15:0 (44.89%), anteiso-C17:0 (6.18%), isoC15:0 (19.38%), and iso-C16:0 (7.39%). Several chemotaxonomic features conform the isolate be a member of genus Halobacillus. The isolate KGW1T contained A1γ meso-Dpm-direct type of peptidoglycan which is different from its phylogenetically closest neighbours. The 16S rRNA gene sequence based phylogenetic analysis also revealed the strain KGW1T was affiliated to the genus Halobacillus and sequence similarity between the isolated strain and the type strains of Halobacillus species were found closest to, H. dabanensis D-8 DSM 18199T (99.08%) and H. faecis IGA7-4 DSM 21559T (99.01%), H. trueperi SL-5 DSM 10404T (98.94%). The in silico DDH showed that the values in a range of 14.2-17.5% with the most closest strain H. dabanensis D-8 DSM 18199T and other type strains of the genus Halobacillus for which whole genome sequence is reported. DNA-DNA relatedness between strain KGW1T and the closest type strain Halobacillus trueperi DSM 10404T was 11.75% (± 1.15). The draft genome sequence includes 3,683,819 bases and comprises of 3898 predicted coding sequences with a G + C content of 46.98%. Thus, the significant distinctiveness supported by phenotypic and genotypic data with its closest neighbors and other closely related species confirm the strain KGW1T to be classified as a novel species within the genus Halobacillus, for which the name Halobacillus marinus sp. nov. is proposed. The type strain is KGW1T (= DSM 29522 = JCM 30443).