Alkalimonas mucilaginosa sp. nov. and Alkalimonas cellulosilytica sp. nov. isolated from alkaline Lonar lake, India.

Two alkaliphilic, Gram-stain-negative bacterial strains (MEB004T and MEB108T) were isolated from water samples collected from Lonar lake, India. The phylogenetic analysis of their 16S rRNA gene sequences showed the highest similarity to A. delamerensis DSM 18314T (98.4%), followed by A. amylolytica DSM 18337T and A. collagenimarina JCM 14267T (97.9%). The genome sizes of strains MEB004T and MEB108T were determined to be 3,858,702 and 4,029,814 bp, respectively, with genomic DNA G + C contents of 51.4 and 51.9%. Average Nucleotide Identity, DNA-DNA Hybridization and Amino Acid Identity values between strains (MEB004T and MEB108T) and A. amylolytica DSM 18337T were (82.3 and 85.5), (25.0 and 29.2) and (86.7 and 90.2%). Both novel strains produced industrially important enzymes, such as amylase, lipase, cellulase, caseinase, and chitinase at pH 10 evidenced by the genomic presence of carbohydrate-active enzymes encoding genes. Genomic analyses further identified pH tolerance genes, affirming their adaptation to alkaline Lonar Lake. Dominant fatty acids were Summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), C16:0, Summed feature 3, Sum In Feature 2 and C12:0 3OH. The prevalent polar lipids included phosphatidyl ethanolamine, phosphatidyl glycerol, and diphosphatidyl glycerol. The major respiratory quinone was ubiquinone-8. Based on the polyphasic data, we propose the classification of strains MEB004T and MEB108T as novel species within the genus Alkalimonas assigning the names Alkalimonas mucilaginosa sp. nov. and Alkalimonas cellulosilytica sp. nov., respectively. The type strains are MEB004T (= MCC 5208T = JCM 35954T = NCIMB 15460T) and MEB108T (= MCC 5330T = JCM 35955T = NCIMB 15461T).

Taxogenomics of Alkalihalobacterium chitinilyticum sp. nov.: an alkaliphilic chitin degrading bacterial strain isolated from Lonar Lake, India, with potential biotechnological applications.

A novel chitin degrading alkaliphilic bacterial strain (MEB 203 T) was isolated from sediment collected from Lonar lake, India. The strain exhibited its maximum growth at a temperature of 37 °C, with an optimal pH of 10 and a NaCl concentration of 2%. 16S rRNA gene based phylogenetic tree showed that strain was closely related to Alkalihalobacterium elongatum MCC 2982 T (98.64% similarity) followed by A. alkalinitrilicum DSM 22532 T (97.84% similarity). The genome size was 4.9 Mb with DNA G + C content of 37.7%. The dDDH value between strain MEB 203 T and A. elongatum MCC 2982 T was 26.4 ± 2.4% while OrthoANI value was 82.1%. Genome analysis revealed the presence of genes responsible for L-ectoine and cation/proton antiporter which may facilitate growth of strain in alkaline-saline habitat of Lonar lake. Strain MEB 203 T was able to utilize complex sugars such as chitin, cellulose, and starch as a carbon source at alkaline conditions which was also corroborated from the genomic presence of carbohydrate active enzymes (CAZymes). It was also able to produce biotechnologically important enzymes such as lipases and proteases which were stable at pH (9-10). The bacterium is majorly composed of C15:0 iso, C16:0 iso, and C17:0 iso (> 10%) fatty acids while diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and unidentified phospholipid (PL3) were identified as the predominant polar lipids. Based on differential physiological, biochemical, and genomic features of strain MEB 203 T, a novel species Alkalihalobacterium chitinilyticum sp. nov. (Type strain MEB 203 T = MCC 3920 T = NCIMB 15407 T = JCM 35078 T) is proposed.

Genomic insights of an alkaliphilic bacterium Halalkalibacter alkaliphilus sp. nov. isolated from an Indian Soda Lake.

An alkaliphilic, Gram-stain-positive, non-motile, rod-shaped, and spore forming bacterial strain (MEB205T) was isolated from sediment sample collected from Lonar lake, India. The strain grew optimally at pH 10, NaCl concentration of 3.0% at 37 °C. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain MEB205T belonged to the genus Halalkalibacter in the family Bacillaceae and shared the highest sequence similarity with H. okhensis Kh10-101T (98.9%) followed by H. wakoensis N-1 T (98.7%). The assembled genome of strain MEB205T has a total length of 4.8 Mb with a G + C content of 37.8%. The dDDH and OrthoANI values between strain MEB205T and H. okhensis Kh10-101 T were 29.1% and 84.3%, respectively. Furthermore, the genome analysis revealed the presence of antiporter genes (nhaA and nhaD) and L-ectoine biosynthesis gene required for survival of the strain MEB205T in alkaline-saline habitat. The major fatty acid was C15:0 anteiso, C16:0 and C15:0 iso (> 10.0%). Diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine were the major polar lipids. meso-diaminopimelic acid was diagnostic diamino acid for cell wall peptidoglycan. Based on the polyphasic taxonomic studies, strain MEB205T represent a novel species of the genus Halalkalibacter for which the name Halalkalibacter alkaliphilus sp. nov. (Type strain MEB205T = MCC 3863 T = JCM 34004 T = NCIMB 15406 T) is proposed.

Corrigendum: Alkalihalobacterium elongatum gen. nov. sp. nov.: An Antibiotic-Producing Bacterium Isolated From Lonar Lake and Reclassification of the Genus Alkalihalobacillus Into Seven Novel Genera.

[This corrects the article DOI: 10.3389/fmicb.2021.722369.].

Alkalihalobacterium elongatum gen. nov. sp. nov.: An Antibiotic-Producing Bacterium Isolated From Lonar Lake and Reclassification of the Genus Alkalihalobacillus Into Seven Novel Genera.

A Gram-stain positive, long, rod-shaped, motile, and spore-forming bacterium (MEB199T) was isolated from a sediment sample collected from Lonar Lake, India. The strain was oxidase and catalase positive. The strain grew optimally at pH 10, NaCl concentration of 3.5% at 37°C. The major fatty acids were iso-C15:0, iso-C16:0, anteiso-C15:0, and iso-C17:0. The peptidoglycan contained meso-diaminopimelic acid (meso-DAP). Phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol were the major polar lipids of MEB199T. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain MEB199T belonged to the family Bacillaceae and exhibited a distinctive position among the members of the genus Alkalihalobacillus (Ahb.). Strain MEB199T shared the highest 16S rRNA gene sequence similarity with Alkalihalobacillus alkalinitrilicus ANL-iso4T (98.36%), whereas with type species Ahb. alcalophilus DSM 485T, it is 94.91%, indicating that strain MEB199T is distinctly related to the genus Alkalihalobacillus. The G + C content of genomic DNA was 36.47 mol%. The digital DNA-DNA hybridization (dDDH) (23.6%) and average nucleotide identity (ANI) (81%) values between strain MEB199T and Ahb. alkalinitrilicus ANL-iso4T confirmed the novelty of this new species. The pairwise identity based on the 16S rRNA gene sequence between the species of genus Alkalihalobacillus ranges from 87.4 to 99.81% indicating the heterogeneity in the genus. The different phylogenetic analysis based on the genome showed that the members of the genus Alkalihalobacillus separated into eight distinct clades. The intra-clade average amino acid identity (AAI) and percentage of conserved proteins (POCP) range from 52 to 68% and 37 to 59%, respectively, which are interspersed on the intra-genera cutoff values; therefore, we reassess the taxonomy of genus Alkalihalobacillus. The phenotypic analysis also corroborated the differentiation between these clades. Based on the phylogenetic analysis, genomic indices, and phenotypic traits, we propose the reclassification of the genus Alkalihalobacillus into seven new genera for which the names Alkalihalobacterium gen. nov., Halalkalibacterium gen. nov., Halalkalibacter gen. nov., Shouchella gen. nov., Pseudalkalibacillus gen. nov., Alkalicoccobacillus gen. nov., and Alkalihalophilus gen. nov. are proposed and provide an emended description of Alkalihalobacillus sensu stricto. Also, we propose the Ahb. okuhidensis as a heterotypic synonym of Alkalihalobacillus halodurans. Based on the polyphasic taxonomic analysis, strain MEB199T represents a novel species of newly proposed genus for which the name Alkalihalobacterium elongatum gen. nov. sp. nov. is proposed. The type strain is MEB199T (= MCC 2982T, = JCM 33704T, = NBRC 114256T, = CGMCC 1.17254T).

Nitrincola tapanii sp. nov., a novel alkaliphilic bacterium from An Indian Soda Lake.

A novel Gram-stain-negative bacterial strain designated as MEB193T was isolated from a sediment sample collected from Lonar Lake, India. The cells were motile, non-spore-forming and rod-shaped. The strain was oxidase- and catalase-positive. It grew optimally at pH 9.0 and at 1 % (w/v) NaCl concentration at 30 °C. Based on 16S rRNA gene sequence similarity, MEB193T belongs to genus Nitrincola, with Nitrincola alkalilacustris ZV-19T (95.89 %) and Nitrincola lacisaponensis 4CAT (95.87 %) as its closest neighbours. The major fatty acid was summed feature 8 comprising C18:1ω7c/C18:1ω6c (52 %) followed by C16 : 0 (25 %). Phosphatidylglycerol (PG) and diphosphatidylglycerol (DPG) were present as the major polar lipids. The draft genome obtained in this study was 2 793 747 bp and the G+C content was 50.79 mol%. Average nucleotide identity (71.76 %) and DNA-DNA hybridization (<20 %) values between strain MEB193T and Nitrincola lacisaponensis 4CAT confirmed the novelty of this new species. Based on phenotypic including chemotaxonomic and genotypic characterization data, strain MEB193T represents a new species of the genus Nitrincola for which the name Nitrincola tapanii sp. nov. is proposed. The type strain is MEB193T (=MCC 2863T=JCM 31570 T=KCTC 52390 T).

Nitrincola alkalisediminis sp. nov., an alkaliphilic bacterium isolated from an alkaline lake.

Two Gram-stain-negative, aerobic, alkaliphilic bacteria (strains MEB087T and MEB142) were isolated from sediment and water samples, respectively, collected from the alkaline Lonar Lake in Maharashtra, India. Strains MEB087T and MEB142 shared 99.8 % 16S rRNA gene sequence similarity and were 85 % related on the basis of DNA-DNA hybridization. The 16S rRNA gene sequences of both strains showed close relationship with the genus Nitrincola, and their closest neighbour was Nitrincola lacisaponensis 4CAT with 97.7 % sequence similarity. MEB087T and MEB142 exhibited only 45 % and 54 % DNA-DNA relatedness, respectively, with Nitrincola lacisaponensis DSM 16316T. Both strains were asporogenous, short, non-motile rods capable of utilizing a limited range of organic acids as sole carbon and energy sources. They were oxidase- and catalase-positive, able to reduce nitrate and nitrite; but unable to degrade DNA, urea, gelatin, casein or starch. They grew optimally at pH 9.5 (tolerating up to pH 11) and could withstand up to 0.6 M NaCl. The predominant cellular fatty acids were summed feature 8 comprising C18 : 1ω7c/C18 : 1ω6c (47-49 %) followed by summed feature 3 comprising C16 : 1ω7c/C16 : 1ω6c (28-32 %). The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. The DNA G+C content was 49.3-49.7 mol%. On the basis of the phylogenetic analysis and chemotaxonomic characteristics, strains MEB087T and MEB142 represent a novel species in the genus Nitrincola, for which the name Nitrincola alkalisediminis sp. nov. is proposed. The type strain is MEB087T ( = KCTC 42948T = JCM 19317T) with MEB142 ( = KCTC 42949 = JCM 19318) as an additional strain.