Streptomyces Diastaticus Research Articles

Actinobacteria mediated synthesis of silver nanoparticles using Streptomyces diastaticus and their biological efficacy against human vector mosquitoes and agricultural insect pests

Mosquitoes and agricultural insects were harmful to both public health and crops. Prolonged use of synthetic chemical pesticides for insect pest control was associated with resistance development and negative human and ecological effects. This study investigates the larvicidal activity of Ag-NPs produced using Streptomyces diastaticus against Culex quinquefasciatus, Anopheles stephensi, Aedes aegypti, Spodoptera litura, and Plutella xylostella. UV-Vis spectroscopy, XRD, SEM-EDX, FT-IR, TEM, DLS, and Zeta potential analysis were used to characterize the biosynthesized Ag-NPs. The results showed that biosynthesized Ag-NPs were more toxic to, A. stephensi, C. quinquefasciatus, and A. aegypti with LC50 values of 15.70, 11.10, and 17.53 µg/mL, respectively. Biosynthesized Ag-NPs exhibited the highest antifeedant effects against S. litura (88.73 %) and P. xylostella (92.36 %) at 50 µg/mL concentration. The mortality bioassay revealed considerable larvicidal activity against S. litura and P. xylostella larvae at greater doses of 50 µg/mL, 80.67 and 84.05 %, respectively. Morphogenetic variations were observed on treated larvae of C. quinquefasciatus, A. aegypti, and A. stephensi, S. litura, and P. xylostella with Ag-NPs. In the present investigation the impact of Ag-NPs on the antioxidant enzymes (SOD, CAT, and GPx), the activity of each enzyme were dramatically increased with higher concentrations resulting in significant differences. The study also found that Ag-NPs inhibited the detoxifying enzyme (GST and AChE) activity of target organisms. The histological structure of the treated and untreated larvae revealed structural changes between the midgut epithelial cells. As a consequence of these findings, the S. diastaticus-mediated Ag-NPs serve as a possible, eco-friendly biopesticide and alternative to synthetic chemicals for the control of human vector mosquitoes and destructive arthropod insect pests.

Screening and production of bacterial amylase from different Strepomyces species

The enzymes from the microbial sources are more stable and can be obtained from cheap sources. Amylase is most important industrial enzyme which is widely used in food and biofuel industries. Streptomyces is a group of bacteria belongs to largest genus of Actinobacteria and commonly isolated from the soil. Streptomyces species have been widely used as microbial cell in industries for production of antibiotics. For the production of antibiotics, Streptomyces species are frequently used as microbial cells. This study reported that screening and production of amylase from different Streptomyces species i.e. Streptomyces microflavus, Streptomyces cyaneus and Streptomyces diastaticus. The cultures were screened for the production of amylase by starch agar plate assay. Result shown the Streptomyces diastaticus was better amylase enzyme producer compare to Streptomyces microflavus and Streptomyces cyaneus. Streptomyces diastaticus gives maximum starch hydrolysis by showing clear zone (4.4 mm), enzyme production (1.60 U/ml) and protein concentration (2.25 mg/ml).

Characterization of the surugamide biosynthetic gene cluster of TUA-NKU25, a Streptomyces diastaticus strain isolated from Kusaya, and its effects on salt-dependent growth.

Kusaya, a traditional Japanese fermented fish product, is known for its high preservability, as it contains natural antibiotics derived from microorganisms, and therefore molds and yeasts do not colonize it easily. In this study, the Streptomyces diastaticus strain TUA-NKU25 was isolated from Kusaya, and its growth as well as the production of antibiotics were investigated. Strain TUA-NKU25 showed advantageous growth characteristics in the presence, but not in the absence, of sodium chloride (NaCl). Antimicrobial assay, high-performance liquid chromatography, and electrospray ionization-mass spectrometry analysis showed that this strain produced surugamide A and uncharacterized antimicrobial compound(s) during growth in the presence of NaCl, suggesting that the biosynthesis of these compounds was upregulated by NaCl. Draft genomic analysis revealed that strain TUA-NKU25 possesses a surugamide biosynthetic gene cluster (sur BGC), although it is incomplete, lacking surB/surC. Phylogenetic analysis of strain TUA-NKU25 and surugamide-producing Streptomyces showed that sur BGC formed a clade distinct from other known groups.

Feature-based molecular networking-guided discovery of siderophores from a marine mesophotic zone Axinellida sponge-associated actinomycete Streptomyces diastaticus NBU2966

Ten siderophore-related compounds were isolated by MS/MS-based molecular networking from a mesophotic sponge-associated Streptomyces diastaticus NBU2966. Among them, there were three kinds of siderophores including three undescribed phenol/thiazoline-type compounds, thiazostatin C, methyl thiazostatin B, and pulicatin J, one undescribed phenol/oxazoline-type compound, spoxazomicin E, and one undescribed phenol/oxazoline/thiazoline-type compound, streptochelin A, as well as five known compounds. Pulicatin J and spoxazomicin E were both isolated as a pair of inseparable epimers. The planar structures of all these compounds were determined based on the detailed NMR and HRESIMS spectroscopic analysis, and the absolute configurations of them were assigned by ECD and NMR quantum chemical calculations. Additionally, in vitro antibacterial activities, inhibition of MRSA biofilm formation, and neuroprotective activity for these compounds were tested.

Characterization of the diastaphenazine/izumiphenazine C biosynthetic gene cluster from plant endophyte Streptomyces diastaticus W2.

SummaryTwo phenazine compounds, diastaphenazine and izumiphenazine C, with complex structures and promising antitumour activity have been isolated from the plant endophytic actinomycete Streptomyces diastaticus W2. Their putative biosynthetic gene cluster (dap) was identified by heterologous expression and gene knockout. There are twenty genes in the dap cluster. dap14‐19 related to shikimic pathway were potentially involved in the precursor chorismic acid biosynthesis, and dapBCDEFG were confirmed to be responsible for the biosynthesis of the dibenzopyrazine ring, the nuclear structure of phenazines. Two transcriptional regulatory genes dapR and dap4 played the positive regulatory roles on the phenazine biosynthetic pathway. Most notably, the dimerization of the dibenzopyrazine ring in diastaphenazine and the loading of the complex side chain in izumiphenazine C could be catalysed by the cyclase homologous gene dap5, suggesting an unusual modification strategy tailoring complex phenazine biosynthesis. Moreover, metabolite analysis of the gene deletion mutant strain S. albus::23C5Δdap2 and substrate assay of the methyltransferase Dap2 clearly revealed the biosynthetic route of the complex side chain in izumiphenazine C.

ANTIBACTERIAL ACTIVITY Streptomyces spp. ENDHOPHYTIC TO Staphylococcus aureus AND Eschericia coli

The objective of this research was to known antibacterial activity of 4 endhophytic Streptomyces namely Streptomyces griseorubiginosus, Streptomyces vellosus, Streptomyces diastaticus and Streptomyces griseoruber against Staphylococcus aureus and Eschericia coli and measure the biggest antibacterial activity which is indicate by clear zone diameter. Antibacterial activity by using agar plug diffusion method showed that Streptomyces vellosus has capable to inhibits growth of Staphylococcus aureus but not with Eschericia coli and Streptomyces diastaticus has capable to inhibits growth of Eschericia coli but not with Staphylococcus aureus. The clear zone of Streptomyces vellosus and Streptomyces diastaticus are 5,18 mm and 7 mm respectively. Streptomyces griseoruber and Streptomyces griseorubiginosus can’t grow during isolation. Keyword: Streptomyces vellosus, Streptomyces diastaticus, agar plug diffusion method

ANTIBACTERIAL ACTIVITY Streptomyces spp. ENDHOPHYTIC TO Staphylococcus aureus AND Eschericia coli

The objective of this research was to known antibacterial activity of 4 endhophytic Streptomyces namely Streptomyces griseorubiginosus, Streptomyces vellosus, Streptomyces diastaticus and Streptomyces griseoruber against Staphylococcus aureus and Eschericia coli and measure the biggest antibacterial activity which is indicate by clear zone diameter. Antibacterial activity by using agar plug diffusion method showed that Streptomyces vellosus has capable to inhibits growth of Staphylococcus aureus but not with Eschericia coli and Streptomyces diastaticus has capable to inhibits growth of Eschericia coli but not with Staphylococcus aureus. The clear zone of Streptomyces vellosus and Streptomyces diastaticus are 5,18 mm and 7 mm respectively. Streptomyces griseoruber and Streptomyces griseorubiginosus can’t grow during isolation. Keyword: Streptomyces vellosus, Streptomyces diastaticus, agar plug diffusion method

Veratraldehyde as a food additive produced by the marine isolate Streptomyces diastaticus LC360811

Twelve actinomycete isolates were derived from 6 sediment samples collected from the Red Sea coast, Egypt. The marine isolate, RS10, the most potent candidate, proved its capability to produce antimicrobial compound(s) that strongly inhibit both Gram-positive and Gram-negative bacteria. RS10 was identified by 16S rRNA gene sequencing as Streptomyces diastaticus LC360811. Culture supernatant of S. diastaticus LC360811 was extracted using n-butanol and tested against bacterial indicator strains using the disc diffusion method. The crude extract was partially purified using column chromatography and assessed for antimicrobial activity. Fraction no. 35 showed the highest antimicrobial activity against Acinatobacter sp. (34 mm), Pseudomonas aeruginosa ATCC 9027 (30 mm), Candida albicans (26 mm), Staphylococcus aureus ATCC 47077 (25 mm), Aeromonas hydrophila (23 mm),Streptococcus iniae (22 mm)and Borditella sp. (18 mm). This active fraction was then identified as veratraldehyde using spectroscopic techniques: GC-MS-MS and HPLC (diode array detector) and compared with the authentic reference. Veratraldehyde as a natural preservative proved to be a good candidate to preserve the flavor of juice, yogurt, and fermented milk and was able to control the growth of bacterial pathogens compared to the non-treated products till day fourteen.

Streptomyces diastaticus isolated from the marine crustacean Portunus sanguinolentus with potential antibiofilm activity against Candida albicans.

Biofilm-mediated multidrug resistance has turned into major challenge for the treatment of C. albicans infections. In the present study, actinomycetes (SS5) isolated from marine crustacean were investigated for their ability to inhibit C. albicans biofilm formation. Cultural, morphological and 16S rRNA analysis revealed that the isolated strain was Streptomyces diastaticus. Ethyl acetate bioactive fractions (6µgmL-1) from SS5 showed potent antibiofilm activity against C. albicans. Light microscopic and CLSM analysis further substantiated the antibiofilm activity of the bioactive fraction against C. albicans. The bioactive fraction was subjected to FTIR and GC-MS for characterization. From GC-MS analysis, the presence of 31 compounds were revealed, among which the alkanes are predominantly present. Hence, further investigation for the potential of these bioactive compounds against C. albicans biofilm will help in the identification of promising candidate for the prevention of biofilm-mediated infection.

L-arginine amidinohydrolase by a new Streptomyces isolate: Screening and statistical optimized production using response surface methodology

Abstract l -arginine amidinohydrolase has been reported as a powerful anticancer agent due to its mediating l -arginine starvation. The aim of this study was to screen a pool of 30 actinomycetes belonging to the species Streptomyces in order to select the potent argininolytic one. Streptomyces strains isolated from soil were firstly functional screened based on their argininolytic capacities by a qualitative assay based on a pH indicator substrate (phenol red). The second screening was the quantitative evaluation of the l -arginine amidinohydrolase activity via a Ninhydrine assay. The measurement was performed on two different cell preparations, involving the extracellular compartment and the cell lysates extract. Among the tested isolates, a strain coded as MAM5 produced extracellular and intracellular l -arginase simultaneously under the same culture conditions. The isolate was identified as Streptomyces diastaticus MAM5 by phylogenetic analysis of the 16S rRNA sequence. Optimization and validation of different process parameters via Box-Behnken design were done for maximum production of extracellular and intracellular l -arginase. Collectively, culturing in shake flasks with the optimized medium resulted in a 4.5-fold and 3.5-fold increase in extra- and intracellular l -arginase production, respectively, when compared to screening medium. Additionally, l -arginase was partially purified using heat treatment and precipitation with cold ethanol, and the enzyme exhibiting 3.2-fold and 3.1-fold for extra- and intracellular form respectively, in comparison with the crude enzyme. Both forms of l -arginase had maximum activity in alkaline pH (8.5–9) at 40–50 °C with ionic stability within pH range 8–10 and thermal stability below 60 °C.

Two new phenazine metabolites with antimicrobial activities from soil-derived Streptomyces species.

Two new phenazine metabolites, 6-hydroxyphenazine-1-carboxamide (1) and methyl 6-carbamoylphenazine-1-carboxylate (2), were isolated from a soil-derived Streptomyces diastaticus subsp. ardesiacus strain YIM PH20246, and their structures were elucidated by extensive spectroscopic data analysis. The antimicrobial activities of the isolates were assayed. Compound 1 showed moderate antifungal and antibacterial activities against Fusarium oxysporum (ATCC 7808), Fusarium solani (ATCC 36031) and Plectosphaerella cucumerina (local isolate), Staphylococcus aureus (ATCC 25923) and Staphylococcus albus (ATCC 10231), respectively. Compound 2 exhibited moderate antifungal and antibacterial activities against F. oxysporum, F. solani, and Phoma herbarum, S. aureus, S. albus, and Bacillus subtilis, respectively.

UTILIZATION OF AGROINDUSTRIAL BY-PRODUCTS AS SUBSTRATE IN ENDOGLUCANASE PRODUCTION BY Streptomyces diastaticus PA-01 UNDER SUBMERGED FERMENTATION

Abstract Endoglucanase production under submerged fermentation was studied using sugarcane bagasse (SCB) and oat bran (OB) as carbon source and corn steep liquor (CSL) as nitrogen source, in different concentrations using factorial design. Streptomyces diastaticus PA-01, isolated from a soil cave in Brazil, was selected as cellulolytic strain. The results after experimental validation showed that a medium containing 2.4% (w/v) SCB and 1.3% (w/v) CSL led to the highest production, 1,180.3 U.L-1 of endoglucanase, after the 5th-day. A good level of endoglucanase (1,039.3 U.L-1) was obtained after the 4th-day when 2.0% (w/v) OB and 1.65% (w/v) CSL were used. The pH and temperature profiles showed thermoacidophilic endoglucanase activity, with 70% of maximum activity at 50oC, after 4 hours of pre-incubation. This is the first report on endoglucanase production by S. diastaticus PA-01 in the presence of SCB and OB. The strong positive effects of some metal ions (Zn2+, Mn2+ and Ba2+) on endoglucanase activity when this strain was grown on OB is an interesting biochemical characteristic for future biotechnological applications.

Angucycline Glycosides from Mangrove-Derived Streptomycesdiastaticus subsp. SCSIO GJ056.

Nine new angucycline glycosides designated urdamycins N1–N9 (1–9), together with two known congener urdamycins A (10) and B (11), were obtained from a mangrove-derived Streptomyces diastaticus subsp. SCSIO GJ056. The structures of new compounds were elucidated on the basis of extensive spectroscopic data analysis. The absolute configurations of 6–9 were assigned by electronic circular dichroism calculation method. Urdamycins N7 (7) and N8 (8) represent the first naturally occurring (5R, 6R)-angucycline glycosides, which are diastereomers of urdamycins N6 (6) and N9 (9), respectively.

Marine actinomycetes as bioremediators in Penaeus monodon rearing system

Actinomycetes (277 Nos) isolated from marine environment and shrimp culture pond sediments were tested for hydrolytic enzyme production and biogranulation property. Potential isolates were screened for their efficacy in bioremediation of shrimp culture system. Based on the BOD reduction efficiency and water quality parameters, five actinomycete isolates viz., Streptomyces coelicoflavus (A6), Streptomyces diastaticus (A44), Nocardiopsis alba (A55), Streptomyces parvus (A56) and Streptomyces champavatii (R32) were subjected for tertiary screening in Penaeus monodon larval rearing system and the animals were challenged with white spot syndrome virus (WSSV). The bioremediating effect of actinomycete treatments were assessed by analysing the expression profile of five antimicrobial peptide (AMP) genes viz., anti-lipopolysaccharide factor (ALF), crustin-2, crustin-3, penaeidin-3 and penaeidin-5 and eight immune genes viz., alpha-2-macroglobulin (α-2-M), astakine, glutathione-S-transferase, haemocyanin, peroxinectin, pmCathepsinC, prophenol oxidase (proPO) and Rab-7. Expression of eight WSSV genes viz., DNA polymerase, endonuclease, protein kinase, immediate early gene, latency related gene, ribonucleotide reductase, thymidine kinase and VP28 were also analyzed to detect the presence and intensity of viral infection in the experimental animals post-challenge. Theapplication of consortia (1 g/5 L water) yields better results in terms of significant reduction in BOD of shrimp rearing system showing the bioremediation potential of the marine actinomycete strains. The application of marine actinomycetes viz., Streptomyces coelicoflavus (A6), Streptomyces diastaticus (A44), Nocardiopsis alba (A55), Streptomyces parvus (A56) and Streptomyces champavatii (R32) in granulated form were found to be potential bioremediators in shrimp rearing system.

Biodegradation of cypermethrin metabolites using terrestrial actinobacterium, Streptomyces diastaticus (PA2) and its GC-MS analysis

Biodegradation of cypermethrin metabolites using terrestrial actinobacterium, Streptomyces diastaticus (PA2) and its GC-MS analysis

Evaluation of antagonistic and plant growth promoting activities of chitinolytic endophytic actinomycetes associated with medicinal plants against Sclerotium rolfsii in chickpea.

To evaluate the potential of chitinolytic endophytic Actinomycetes isolated from medicinal plants in order to diminish the collar rot infestation induced by Sclerotium rolfsii in chickpea. Sixty-eight chitinolytic endophytic Actinomycetes were recovered from various medicinal plants and evaluated for their chitinase activity. Among these isolates, 12 were screened for their plant growth promoting abilities and antagonistic potential against Sc.rolfsii. Further, these isolates were validated invivo for their ability to protect chickpea against Sc.rolfsii infestation under greenhouse conditions. The isolates significantly (P<0·05) increased the biomass (1·2-2·0 fold) and reduced plant mortality (42-75%) of chickpea. On the basis of 16S rDNA profiling, the selected antagonistic strains were identified as Streptomyces diastaticus, Streptomyces fradiae, Streptomyces olivochromogenes, Streptomyces collinus, Streptomyces ossamyceticus and Streptomyces griseus. This study is the first report of the isolation of endophytic Actinomycetes from various medicinal plants having antagonistic and plant growth promoting abilities. The isolated species showed potential for controlling collar rot disease on chickpea and could be useful in integrated control against diverse soil borne plant pathogens. Our investigation suggests that endophytic Actinomycetes associated with medicinal plants can be used as bioinoculants for developing safe, efficacious and environment-friendly biocontrol strategies in the near future.

Evaluation of airborne Actinomycetes at waste application facilities

This study aimed to evaluate airborne meso–and–thermophilic actinomycete concentrations and their types at a wastewater treatment plant and a biosolid landfill, in Egypt. Air samples were collected at 200m upwind, and onsite and 300m downwind by using liquid impinger sampler, calibrated to draw 12.5L/min, for 20 minutes. The concentrations ranged between 0.0–7 360CFU/m3 for mesophilic, and 106–586CFU/m3 for thermophilic actinomycetes. Airborne actinomycete concentrations exceeded the suggested occupational exposure limit value of 100CFU/m3. No significant correlations were found between actinomycete concentrations onsite and 300m downwind distance. At the biosolid landfill, upwind thermophilic actinomycetes significantly differed (p<0.05) from onsite and 300m downwind. A total of 40 and 69 airborne actinomycete isolates belonging to 8 genera were identified at the wastewater treatment plant and biosolid landfill. Streptomyces were the dominant actinomycete species. Streptomyces diastaticus, Pseudonocardia compacta and Catellatospora ferruginea were only detected at the biosolid landfill site. Meso–and–thermophilic actinomycetes positively associated with relative humidity, and wind speed positively correlated with onsite thermophilic actinomycetes (r=0.65) at the biosolid landfill. Temperature showed negative effect on survivability of mesophilic actinomycetes (r=-0.8) onsite of the wastewater treatment plant. Waste application facilities increase actinomycete concentrations onsite which may consequently deteriorate air quality in the nearby areas.

Oligomycins A and C, major secondary metabolites isolated from the newly isolated strain Streptomyces diastaticus

During the screening program for fungicides, one actinomycete strain ECO 00047 was isolated with the potential activity against fungus. According to the morphology and analysis of the nucleotide sequence of the 16S rRNA gene (1500 bp) this isolate was identified as Streptomyces diastaticus. The active compounds were separated by silica gel column chromatography, Sephadex LH-20 gel filtration and then purified by flash chromatography on C18 (20-45 microm). The chemical structure of the bioactive compounds I and II were elucidated, based on the spectroscopic data of MS, IR, UV, 1H-NMR, 13C-NMR and X-ray single crystal diffraction analysis. Compounds I and II were identical with oligomycins A and C, the macrolide antibiotics which have been known to be produced by Streptomyces diastatochromogenes, S. libani and S. avermitilis. The two compounds exhibited a strong activity against Aspergillus niger, Alternaria alternata, Botrytis cinerea and Phytophthora capsici but no activity toward bacteria. Although the two above antibiotics were known, their isolation has so far not been reported from S. diastaticus.

A Tailoring Activity Is Responsible for Generating Polyene Amide Derivatives in Streptomyces diastaticus var. 108

We recently characterized rimocidin B (3b) and CE-108B (4b) as two polyene amides with improved pharmacological properties, produced by genetically modified Streptomyces diastaticus var. 108. In this work, genetic and biochemical analysis of the producer strain show that the two amides are derived from the parental polyenes rimocidin (3a) and CE-108 (4a) by a post-PKS modification of the free side chain carboxylic acid. This modification is mediated by an amidotransferase activity operating after the biosynthesis of rimocidin (3a) and CE-108 (4a) are completed. Two polyenes, intermediates of the biosynthetic pathway of rimocidin (3a) and CE-108 (4a), were also isolated and shown to have some improved pharmacological properties compared with the final products.

Construction of double-copy glucose isomerase gene engineering strain of Streptomyces diastaticus by homologous recombination.

The plasmid pUT for homologous recombination was constructed by the insertion of the 1.1-kb thiostrepton resistance ( tsr(R)) gene into the E. coli plasmid pUB1-GI1. Plasmid pUTK was produced through ligating the cleaved plasmid pUT by KpnI. After pUT and pUTK were introduced into Streptomyces diastaticus No.7 strain M1033 (SM33) by protoplast transformation, a series of tsr(R) transformants were obtained, further based on enzyme assays. These results for polymerase chain reaction (PCR), DNA sequencing, restriction enzyme digestion, and recovery of cloned fragments from the transformant chromosome demonstrated the plasmid pUT and pUTK had integrated into the SM33 chromosome in three different patterns of single cross-over by homologous recombination. This directly results in double-copy GI gene in the transformant chromosome, of which one is wild-type GI gene, the other mutant GI ( GIG138P, GI1) gene. Among the strains of the three kinds of recombinant patterns, one transformant was chosen and named K1, T2, and T3, respectively. The further identification of the three recombinant strains by PCR, DNA sequencing, restriction enzyme digestion, and Southern hybridization also proved there is a double-copy GI gene within their chromosome. Enzyme activity assay and thermostability analysis indicated that all three engineering strains expressed not only wild-type enzyme but also mutant GI.