Streptomyces cupreus sp. nov., an antimicrobial producing actinobacterium isolated from Himalayan soil.

A novel actinobacterium, designated Streptomyces sp strain PSKA01T, was isolated from a soil sample of Kashmir Himalaya, India (latitude:34˚-01'N; longitude:74˚-47'E; altitude: 5328 ft). The taxonomic position of this strain was revealed by a polyphasic approach. Comparative analysis of the 16S rRNA gene sequences indicated that PSKA01T was closely related to the members of the genus Streptomyces, with the highest similarity to Streptomyces venetus CMU-AB225T (99.3%), Streptomyces levis NBRC 15423 T (99.3%), Streptomyces misionensis DSM 40306 T (99.2%), Streptomyces carpinensis NRRL B-16921 T (99.2%), Streptomyces purpurascens NBRC 13077 T (99.2%), and Streptomyces phaeoluteichromatogenes NRRL 5799 T (99.2%). A phylogenomic tree based on genome sequences of the isolated strains, conferred by Type Strain Genome Server (TYGS), separated them from rest of the Streptomyces. The cell wall contained LL-diaminopimelic acid and the whole-cell hydrolysates were xylose and rhamnose. Major fatty acid methyl esters (FAME) were observed as 16:0 iso and 17:0 anteiso. In addition to the differences in phenotypic characters, the average nucleotide identity (ANI), average amino acid identity (AAI), digital DNA-DNA hybridization (dDDH) values, tetranucleotide frequency correlation coefficient (TETRA), and multilocus sequence analysis (MLSA) between PSKA01T and closely related type strains were considerably lower than the recommended threshold value; therefore, this isolate represents a novel Streptomyces species, for which the name Streptomyces cupreus sp. nov. is proposed. The type strain is PSKA01T (= JCM 33752 T = MCC 4182 T).

Streptomyces himalayensis sp. nov. including Streptomyces himalayensis subsp. himalayensis subsp. nov. and Streptomyces himalayensis subsp. aureolus subsp. nov. isolated from Western Himalaya.

Two novel actinobacteria, strain PSKA28T and PSKA54T were isolated from soil sample of Kashmir-Himalaya, India (latitude 34°-01' N; longitude 74°-47' E; altitude 5328 ft). Polyphasic-taxonomic analysis revealed that these strains belong to the genus Streptomyces. The 16S rRNA gene-sequence similarity of these strain were highest (98.6%) with that of the Streptomyces torulosus NRRLB-3889T. They showed 92.1% gyrB gene-sequence similarity but 92.9 and 94.6% rpoB gene-sequence similarity, respectively, with Streptomyces torulosus NRRLB-3889T. The sequence similarities were significantly lower than the recommended threshold value for novel taxonomic position and the 16S rRNA sequence-based phylogenetic analysis indicated that these two isolates are in distinct clade. Phylogenomic analysis with their genome sequences, conferred by Type Strain Genome Server separated them from rest of the Streptomyces type strains. The cell wall contained LL-diaminopimelic acid and the whole-cell hydrolysate contained glucose and ribose. Major fatty acid methyl esters were 15:0 anteiso, 16:0 iso and 17:0 anteiso. Average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization (dDDH) values as well as evolutionary distance based on multilocus sequence analysis, between closely related type strains and either of the strain PSKA28T or PSKA54T, were considerably lower than the recommended threshold value (< 70% dDDH or < 95-96% ANI/AAI or > 0.007 MLSA distance). Therefore, these isolates represent a novel Streptomyces species, for which we proposed the name Streptomyces himalayensis sp. nov. Genotypic and phenotypic analyses clearly split these strains into two closely related sub-clusters. Based on this, two novel subspecies Streptomyces himalayensis subsp. himalayensis subsp. nov. and Streptomyces himalayensis subsp. aureolus subsp. nov. are proposed, with type strains JCM33755T (= MTCC12933T, MCC4145T) and JCM33759T (= MTCC12938, MCC4180), respectively.

Different soil salinity imparts clear alteration in rhizospheric bacterial community dynamics in rice and peanut.

The rhizospheric microbiome is capable of changing the physio-chemical properties of its own micro-environment and found to be indispensable in the overall health of the hostplant. The interplay between the rhizospheric environment and the microbiota residing therein tune the physiology of the associated plant. In this study, we have determined how the soil properties and the host-plant remains as an important parameter for microbial community dynamics in the rhizosphere of rice and peanut. In addition to check the physio-chemical parameters of the rhizospheric soil, we have also prepared the metagenomic DNA from each rhizospheric soil followed by high-throughput sequencing and sequence analysis to predict the OTUs that represents the community structure. The alpha-diversity of the bacterial community in the RRN sample was highest, while the lowest was in PRS sample. Actinobacteria is the most predominant phylum in PRN, PRS and RRN, whereas Acidobacteria in RRS. We found a clear shift in bacterial community over the rice and peanut rhizosphere and also over these host-rhizospheres from normal and high saline region. The rhizospheric bacterial community composition found to be affected by the close-by environmental factors. Thus, the rhizospheric bacterial community structure is related to both the adjoining soil characters and the type of the hosts.

Lentzea indica sp. nov., a novel actinobacteria isolated from Indian Himalayan-soil.

A novel actinobacterium, strain PSKA42T was isolated from a soil sample of Kashmir Himalaya, India (latitude: 34°-01' N; longitude: 74°-47' E; altitude: 5328 ft). The morphological, biochemical and molecular characteristics predicted that PSKA42T belongs to the genera Lentzea. Using the Ezbiocloud server it has also been manifested that the 16S rRNA gene sequence of PSKA42 was 98.4% similar with Lentzea guizhouensis DHSC013T, Lentzea albidocapillata subsp. violacea IMSNU50388T; 98.26% similar with Lentzea aerocolonigenes NRRLB-3298T, Lentzea albidocapillata subsp. albidocapillata NRRLB-24057T and 98.19% similar with Lentzea californiensis DSM43393T. Whole-genome average nucleotide identity values between PSKA42T and L. guizhouensis DHSC013T, L. albidocapillata subsp. violaera IMSNU50388T, L. aerocolonigenes NRRLB-3298T, L. albidocapillata subsp. albidocapillata NRRLB-24057T, L. californiensis DSM43393T, L. flaviverucosa As40578T were 84.94%, 85.8%, 87.15%, 87.71%, 79.29, and 87.22%, respectively, suggesting that PSKA42T represented a new species. In-silco genome-to-genome distances analysis and MLSA with selected housekeeping genes also indicated that the isolate should be assigned to a new species under the genus Lentzea. Analysis of the whole-cell hydrolysate found mannose and ribose as major sugar while cell wall amino acid was identified as meso-diaminopimelic acid. Major fatty acids (> 5%) were observed as 15:0 iso (23.81%), and 16:0 iso (26.11%). PSKA42 contains polyketide synthase type-I genes but unable to show antimicrobial activity in laboratory conditions. The average GC content in its genome is 68.3%. Further, based on physicochemical, chemotaxonomic markers and molecular analysis, PSKA42T can be assigned as novel species of Lentzea, we proposed Lentzea indica sp nov. The type strain is PSKA42T (JCM 33729T, MTCC 12936T, MCC 4127T).

Effect of different stimuli on twitching behavior of endophytic bacteria isolated from Loranthus sp. Jacq.

Bacteria need to adopt to different behavioral tuning depending on the dynamic eco-physiological conditions they are exposed to. One of these adaptive strategies is the use of motility. Here we report the twitching motility response of four endophytic isolates of Bacillus sp. when exposed to different eco-physiological stimuli like different nutrient sources, and mechanical and chemical antagonists on solid surfaces. These endophytic bacteria were isolated from different parts of a hemiparasite Loranthus sp. Jacq. (Loranthaceae) growing on economically important mango trees. The results show that the twitching motility of these bacteria was more when exposed to organic acids, metals salts (among nutrients) and mechanical shearing (stress) than the other factors. Their motility is not affected by surface lubrication or EPS production, but instead is influenced by shear-sensitive structures and affinity to metal ions. Further molecular studies are needed to elucidate the basis of this twitching behaviour on solid surfaces.

Comprehensive genome analysis of Lentzea reveals repertoire of polymer-degrading enzymes and bioactive compounds with clinical relevance.

The genus Lentzea is a rare group of actinobacteria having potential for the exploration of bioactive compounds. Despite its proven ability to produce compounds with medical relevance, Lentzea genome analysis remains unexplored. Here we show a detailed understanding of the genetic features, biosynthetic gene clusters (BGCs), and genetic clusters for carbohydrate-active enzymes present in the Lentzea genome. Our analysis determines the genes for core proteins, non-ribosomal peptide synthetase condensation domain, and polyketide synthases-ketide synthase domain. The antiSMASH-based sequence analysis identifies 692 BGCs among which 8% are identical to the BGCs that produce geosmin, citrulassin, achromosin (lassopeptide), vancosamine, anabaenopeptin NZ857/nostamide A, alkylresorcinol, BE-54017, and bezastatin. The remaining BGCs code for advanced category antimicrobials like calcium-dependent, glycosylated, terpenoids, lipopeptides, thiopeptide, lanthipeptide, lassopeptide, lingual antimicrobial peptide and lantibiotics together with antiviral, antibacterial, antifungal, antiparasitic, anticancer agents. About 28% of the BGCs, that codes for bioactive secondary metabolites, are exclusive in Lentzea and could lead to new compound discoveries. We also find 7121 genes that code for carbohydrate-degrading enzymes which could essentially convert a wide range of polymeric carbohydrates. Genome mining of such genus is very much useful to give scientific leads for experimental validation in the discovery of new-generation bioactive molecules of biotechnological importance.

An antibacterial compound pyrimidomycin produced by Streptomyces sp. PSAA01 isolated from soil of Eastern Himalayan foothill.

Selective isolation of soil Actinobacteria was undertaken to isolate a new class of antibiotics and bioactive molecules. A Streptomyces sp. PSAA01 (= MTCC 13,157), isolated from soil of Eastern Himalaya foothill was cultivated on a large scale for the production of the antimicrobial SM02. It has been found that the maximum amount of SM02 produced while PSAA01 was grown in ISP-2 medium (pH 7.0) for 7 days at 30 °C in shaking (180 rpm) condition. A significant zone of inhibition against Staphylococcus aureus MTCC 96 has been found with the crude cell-free culture media (50 µL) of 7 days grown PSAA01. After the purification and chemical structural characterization, we found that SM02 is a new antimicrobial having 746 dalton molecular weight. The compound SM02 contains pyrimidine moiety in it and is produced by a species of Streptomyces and thus we have named this antibiotic pyrimidomycin. The antimicrobial spectrum of pyrimidomycin has been found to be restricted in Gram-positive organisms with a MIC of 12 µg/mL. SM02 was found active against Mycobacterium sp. and also multi-drug resistant Gram-positive bacteria with similar potency and found to disrupt the bacterial cell wall. Pyrimidomycin also showed significant impairment in the biofilm formation by S. aureus. Furthermore, pyrimidomycin showed synergy with the most used antibiotic like ampicillin, vancomycin and chloramphenicol. Pyrimidomycin did not have cytotoxicity towards human cell lines indicating its limited activity within bacteria.

Draft Genome Sequence of an Endophytic Micromonospora sp. Strain, ANENR4, Isolated from the Root of a Peanut Plant (Arachis hypogaea).

The genus Micromonospora was found to occur in a diverse range of habitats. Here, we report the genome sequence of an endophytic strain of Micromonospora sp., ANENR4. ANENR4 was isolated from the healthy roots of a peanut (Arachis hypogaea) plant from Egra, West Bengal, India.

Morpho-biochemical and molecular characterization of two new strains of Aspergillus fumigatus nHF-01 and A. fumigatus PPR-01 producing broad-spectrum antimicrobial compounds.

The main objective of the study is to characterize two new strains of Aspergillus fumigatus through morphometric, biochemical, molecular methods, and to evaluate their antimicrobial potentiality. The micro-morphotaxonomy, growth, and metabolic behavior of the strains, nHF-01 and PPR-01, were studied in different growth conditions and compared with standard strain. The molecular characterization was done by sequencing the ncrDNA ITS1-5.8S-ITS2 and D1-D2 domains of the nc 28S rDNA region and compared with a secondary structure-based phylogenetic tree. The secretory antimicrobials and pigments were characterized by TLC, UV-Vis, and FT-IR spectroscopy. Both the strains showed distinct growth patterns in different nutritional media and could assimilate a wide range of carbohydrates with distinctive biochemical properties. The molecular characterization revealed the strains, nHF-01 and PPR-01, as Aspergillus fumigatus (GenBank Accession No. MN190286 and MN190284, respectively). It was observed that the strain nHF-01 produces red to brownish pigments having mild antimicrobial activity while the strain PPR-01 does not represent such transformations. The extractable compounds had a significant antimicrobial potentiality against the human pathogenic bacteria. From this analysis, it can be concluded that the nHF-01 and PPR-01 strains are distinct from other A. fumigatus by their unique characters. Large-scale production and detailed molecular elucidation of the antimicrobial compounds may lead to the discovery of new antimicrobial compounds from these strains.

Streptomyces sp SM01 isolated from Indian soil produces a novel antibiotic picolinamycin effective against multi drug resistant bacterial strains.

A Kashmir Himalayan (India) soil isolate, Streptomyces sp. SM01 was subjected to small scale fermentation for the production of novel antimicrobials, picolinamycin (SM1). The production has been optimized which found to be maximum while incubated in AIA medium (pH 7) for 7 days at 30 °C. Seven days grew crude cell-free culture media (50 µL) showed a larger zone of inhibition against Staphylococcus aureus compared to streptomycin (5 µg) and ampicillin (5 µg). Extraction, purification, and chemical analysis of the antimicrobial component has been proved to be a new class of antibiotic with 1013 dalton molecular weight. We have named this new antibiotic as picolinamycin for consisting picolinamide moiety in the center of the molecule and produced by a Streptomyces sp. In general, the antimicrobial potency of this newly characterized antibiotic found to be higher against Gram-positive organisms than the tested Gram-negative organisms. The MIC of this antimicrobial compound was found to be 0.01 µg/ml for tested Gram-positive organisms and 0.02 to 5.12 µg/ml for Gram-negative organisms. Furthermore, it showed strong growth impairments of several multidrug resistance (MDR) strains, including methicillin-resistant strains of Staphylococci and Enterococci with the MIC value of 0.04 to 5.12 µg/ml and MDR (but methicillin-sensitive) strains of S. aureus with the MIC value of 0.084 µg/ml. It also showed anti-mycobacterial potential in higher concentrations (MIC is 10.24 µg/ml). Picolinamycin however did not show toxicity against tested A549 human cell line indicating that the spectrum of its activity limited within bacteria only.

Visualisation of DCP, a nerve agent mimic, in Catfish brain by a simple chemosensor.

A chemosensor, 3-aminophenol-based rhodamine conjugate (ARC) has been developed for visualisation of diethylchlorophosphate (DCP), mimic of a chemical warfare agent, in Catfish brain. The simple detection of DCP by "turn-on" fluorescence property of the chemosensor makes it unique for easy and rapid in vivo and in vitro detection of DCP with the detection limit of 5.6 nM.

2'-Deoxy-5-(hydroxymethyl)cytidine: estimation in human cancer cells with a simple chemosensor.

A pyrrole-based rhodamine conjugate (CS-1) has been developed and characterized for the selective detection and quantification of 2'-deoxy-5-(hydroxymethyl)cytidine (5hmC) in human cancer cells with a simple chemosensing method.