Isolation of rare Genera of actinomycetes — antibiotic producers from soils using Aloe Arborescens juice

The search for new antibiotics is an urgent problem due to the spread of resistance to existing antibacterial drugs in pathogenic microorganisms. Actinomycetes are producers of a large number of antibiotics used in medicine. Most antibiotics are isolated from actinomycetes of the Streptomyces genus, while rare genera of actinomycetes can be the producers of new antibiotics.The aim of the study is to investigate the effect of the biological substances complex present in aloe juice on the growth stimulation of rare genera of actinomycetes.Material and methods. Objects: samples of sod-podzolic soil and chernozem. The standard method of sowing soil suspensions on oat agar and Gause medium No. 2 was used to isolate actinomycetes. Chemotaxonomic properties were determined using the methods of ascending thin-layer chromatography on a cellulose layer. The generic identity of cultures was determined using Bergey’s manual and materials comparing the composition of cell walls of actinobacteria. DNA PCR with standard 27f and 1492r primers, as well as Sanger sequencing, were performed to study genosystematic features. Antibiotic activity was determined against the test microorganisms: Staphylococcus aureus ИНА 00985 (FDA 209P), Staphylococcus aureus ИНА 00761 (MRSA), Staphylococcus aureus ИНА 00762 (УФ- 2), Micrococcus luteus ATCC 9341, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Saccharomyces cerevisiae ИНА 01042.Results. A total of 527 actinomycete cultures were isolated from samples of sod-podzolic soil and chernozem with the addition of aloe juice; their phylogenetic position was determined. The dominant actinomycetes in the studied soil samples are the representatives of the genus Streptomyces. Bacteria of the genus Micromonospora take the second place by the number of isolated cultures. Rare genera of actinomycetes have also been identified: Nonomuraea, Streptosporangium, Nocardia, Actinomadura, Actinocorallia, Pseudonocardia, Amycolatopsis, Saccharomonospora, Saccharopolyspora, Promicromonospora, Kribbella. It was determined that the isolated cultures possess antibiotic activity against test microorganisms.Conclusion. It is advisable to use aloe juice after subjecting the leaves to biostimulation to isolate actinomycetes from the soil and identify their biodiversity.

THE COMPOSITION OF CELLULAR FATTY ACIDS OF ACTINOBACTERIA FROM THE SURFACES OF BIOLOGICAL GROWTH OF THE ODESA GULF OF THE BLACK SEA

Aim. Determination of fatty acid composition of actinobacteria isolated from the surfaces of biological growth of the Odesa gulf of the Black Sea, and their identification. Methods. The 31 isolated strains of actinobacteria were grown in TSB at 28 ° C and 150 rpm for 72 hours. Fatty acid methyl esters of the studied strains were determined according to the MIS Operating Manual on a gas chromatograph Agilent 7890, identification was performed using the identification system of microorganisms MIDI Sherlock. Results. Using chromatographic analysis of fatty acids, it was found that of the 27 studied strains of actinobacteria were identified to the genus Streptomyces, and the 4 strains - to the genus Nocardiopsis. It was found that the fatty acid profiles of the studied actinobacteria of the genus Nocardiopsis were dominated by fatty acids: 15:0 ANTEISO, 16:0 ISO, 17:0 ANTEISO, 18:1 CIS 9, and the fatty acid profiles of bacteria of the genus Streptomyces - 14:0 ISO, 15:0 ANTEISO, 16:0 ISO, 17:0 ANTEISO. Conclusions. Actinobacteria the surfaces of biological growth of the Odesa gulf of the Black Sea belong to the genera Streptomyces and Nocardiopsis, and their fatty acid profiles are characterized by the dominance of isomers of branched saturated fatty acids.

Challenges to Explore Genus Streptomyces in Ethiopia-A Mini Review

Genus Streptomyces is gram-positive bacteria that grow in various environments. It has plentiful biotechnological attributes on the area of agricultural, bioremediation, biofuel, clinical, food, industrial, medical, pharmaceutical, and veterinary. The aim of the review is to frontward challenges to explore potent Streptomyces species in the case of Ethiopia. There is lack of the extent and quality of the genetic research regarding to genome sequence, bioactive compound discovery, and genetic manipulation. Their functional and structural diversity is not full studied. To find a new Streptomyces species: Culture media formulation and optimization as wells culture-independent method like Next Generation Sequencing approach should incorporate at national level.

Streptomyces Pimoensis sp. nov., Isolated From the Taklimakan Desert in Xinjiang, China

A novel Streptomyces strain, designated TRM 75549T, was separated from a sample of sand in Pimo, Taklimakan desert, Xinjiang, North-West China. Phylogenetic analyses of the 16S rRNA gene sequences placed strain TRM75549T within the genus Streptomyces with the highest similarities to Streptomyces flavoviridis NBRC 12772T (98.76%). The whole-genome average nucleotide identity (ANI) value between strain TRM75549T and S. flavoviridis NBRC 12772T is 88.20%. Digital DNA-DNA hybridization (dDDH) value between strain TRM75549T and S. flavoviridis NBRC 12772T is 44.10%. They are well below the recommended 95-96% and 70% cut-off points for designated species respectively. A multi-locus sequence analysis of five house-keeping genes (atpD, gyrB, recA, rpoB and trpB) and phylogenomic analysis also illustrated that strain TRM75549T should be assigned to the genus Streptomyces. Strain TRM75549T contained MK-9 (H6) and MK-9 (H8) as predominant menaquinones. The diagnostic diamino acid of cell walls was identified as LL-diaminopimelic acid and Meso-diaminopimelic. The whole-cell sugar pattern of strain TRM 75549T consisted of mannose and glucose. The major fatty acids (>5%) were iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:1H, iso-C16:0. The polar lipids were diphosphatidylglycerol, lysophosphatidylglycerol, phosphatidylethanolamine, phospholipids, phosphatidylglycerol, phosphatidylinositol, phosphatiylinositol mannosides and unidentified phospholipids. Strain TRM75549T could be differentiated from S. flavoviridis NBRC 12772T, based on physiological and biochemical characteristics. Based on the data from this polyphasic study presented above, strain TRM75549T is represent ative of a novel species of the genus Streptomyces, for which the name Streptomyces pimoensis sp. nov. is proposed. The type strain is TRM75549T (=CCTCC AA 2020054T=LMG 32221T ).

Isolation and characterization of soil actinobacteria as cellulolytic enzyme producer from Aceh Besar, Indonesia

Fitri L, Bessania MA, Septi N, Suhartono S. 2021. Isolation and characterization of soil actinobacteria as cellulolytic enzyme producer from Aceh Besar, Indonesia. Biodiversitas 22: 5169-5180. Cellulolytic actinobacteria are cellulase-producing bacteria capable of degrading cellulose. This study aimed to isolate, characterize, evaluate the cellulolytic ability, and to determine physiological characterization of soil cellulolytic actinobacteria isolated from the Ujung Pancu area, Aceh Besar. Isolation of actinobacteria from soil samples was performed using serial dilution method on Yeast Malt Agar (YMA) medium. Morphological characterization was carried out by growing isolates on YMA, Oatmeal Agar (OA), and Yeast Starch Agar (YSA) media. Cellulolytic ability was determined by calculating the cellulolytic index (IS) on 1% carboxymethyl cellulose (CMC) medium after adding 0.1% congo red solution. Physiological characterization of cellulolytic actinobacteria tested in this study was salinity, pH, and carbon source in liquid Yeast Malt (liquid YM), and the growth was measured at a wavelength of 581nm. The results showed that a total of nine isolates of actinobacteria were isolated, which belonged to the genus Streptomyces. Cellulolytic test results showed that eight isolates had the ability to degrade cellulose. Isolates AUP-04, AUP-03, and AUP-01 had the highest cellulolytic index value. Physiological characterization results revealed that three isolates had different tolerances for salinity levels, pH, and types of carbon sources. AUP-03 isolate grew well at 10% salinity with an OD value of 0.88, isolate AUP-01 grew at 5% salinity with an OD value of 0.49, whereas isolate AUP-04 grew well on media that did not contain salinity. All three isolates grew well at pH 6 with OD values of 0.93, 1.12, and 1.27. AUP-03 and AUP-01 isolates grew well on media containing dextrose as carbon source with OD values of 0.154 and 0.17, respectively, while isolate AUP-04 grew well on glucose-containing media with an OD value of 0.22.

Streptomyces, Greek Habitats and Novel Pharmaceuticals: A Promising Challenge

Bacteria of the genus Streptomyces produce a very large number of secondary metabolites, many of which are of vital importance to modern medicine. There is great interest in the discovery of novel pharmaceutical compounds derived from strepomycetes, since novel antibiotics, anticancer and compounds for treating other conditions are urgently needed. Greece, as proven by recent research, possesses microbial reservoirs with a high diversity of Streptomyces populations, which provide a rich pool of strains with potential pharmaceutical value. This review examines the compounds of pharmaceutical interest that have been derived from Greek Streptomyces isolates. The compounds reported in the literature include antibiotics, antitumor compounds, biofilm inhibitors, antiparasitics, bacterial toxin production inhibitors and antioxidants. The streptomycete biodiversity of Greek environments remains relatively unexamined and is therefore a very promising resource for potential novel pharmaceuticals.

Multifaceted bioproperties of Streptomyces bacillaris ANS2 isolated from Andaman and Nicobar Islands, India

The rich warehouse called marine actinobacteria has enticed researchers for several decades. The assessment of their metabolites has evidenced several diverse and multiple biological properties vacillating from antibacterial, antituberculosis, antioxidant, anticancer to mosquitocidal. Therefore, the present study has been aimed to reconnoitre the isolation and portrayal of the promising marine actinobacteria named Streptomyces bacillaris ANS2 isolated from the mangrove sediment collected from Andaman and Nicobar islands. The ethyl acetate extract of ANS2 showed broad spectrum activity against various pathogens including Escherichia coli (18.0±0.3 mm), Salmonella paratyphi (16.0±0.6 mm) and Klebsiella pneumonia (14.8±0.7 mm). It showed maximum activity against drug sensitive strain of Mycobacterium tuberculosis (98.2±0.36%) followed by M. tuberculosis MDR strain (97.04±1.32%) and M. tuberculosis H37Rv (91.80±0.45%). The antioxidant activity by DPPH assay of ANS2 showed 29.9±0.38 - 61.35±0.78 % free radical scavenging at 100μg/ml - 500μg/ml as compared to the standard ascorbic acid which showed 96±1.0% at 100μg/ml concentration. The anticancer activity showed 88.12±0.9% and 89.5±0.26% inhibition against HT 29 (colon cancer) and HeLa (cervical cancer) cell line at 1000μg/ml concentration. The maximum larval mortality was recorded on Culex quinquefasciatus(LC50 =1100.134 μg/ml and r2= 0.99) and Aedes aegypti (LC50 =690.620 μg/ml and r2= 0.99).The biochemical, morphological and 16S rRNA gene analysis study clearly confirmed that the promising strain belonged to the genus Streptomyces bacillaris and the phylogenetic tree confirmed it as S. bacillaris. Results showed that S.bacillaris ANS2 showed multidimensional bioproperties.

Antimicrobial Profile of Actinomycin D Analogs Secreted by Egyptian Desert Streptomyces sp. DH7

Egyptian deserts are an underexplored ecological niche, especially the Sinai Peninsula. In our recent study, we explored this extreme environment and shed light on the bioactive capabilities of desert Actinobacteria isolated from Sinai. Fifty desert Actinobacteria were isolated from the Sinai desert using mineral salt media, basal media, and starch casein media. The filtrate of Streptomyces sp. DH 7 displayed a high inhibitory effect against multidrug-resistant Staphylococcus aureus (MRSA) strains. The 16S rDNA sequencing confirmed that isolate DH7 belongs to the genus Streptomyces. The NJ phylogenetic tree showed relatedness to the Streptomyces flavofuscus strain NRRL B-2594 and Streptomyces pratensis strain ch24. The minimum inhibitory concentrations against MRSA were 16 and 32 μg/μL. Chemical investigation of the ethyl acetate extract of Streptomyces sp. DH7 led to the isolation and purification of natural products 1–4. Structure elucidation of the purified compounds was performed using detailed spectroscopic analysis including 1 and 2D NMR, and ESI-MS spectrometry. To the best of our knowledge, this is the first report for the isolation of compounds 1–4 from a natural source, while synthetic analogs were previously reported in the literature. Compounds 3–4 were identified as actinomycin D analogues and this is the first report for the production of actinomycin D analogs from the Sinai desert with an inhibitory effect against MRSA. We indorse further study for this analog that can develop enhanced antimicrobial activities. We confirm that the desert ecosystems in Egypt are rich sources of antibiotic-producing Actinobacteria.