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1.
Article in English | WPRIM | ID: wpr-982716

ABSTRACT

Lysobacter harbors a plethora of cryptic biosynthetic gene clusters (BGCs), albeit only a limited number have been analyzed to date. In this study, we described the activation of a cryptic polyketide synthase (PKS)/nonribosomal peptide synthetase (NRPS) gene cluster (lsh) in Lysobacter sp. DSM 3655 through promoter engineering and heterologous expression in Streptomyces sp. S001. As a result of this methodology, we were able to isolate two novel linear lipopeptides, lysohexaenetides A (1) and B (2), from the recombinant strain S001-lsh. Furthermore, we proposed the biosynthetic pathway for lysohexaenetides and identified LshA as another example of entirely iterative bacterial PKSs. This study highlights the potential of heterologous expression systems in uncovering cryptic biosynthetic pathways in Lysobacter genomes, particularly in the absence of genetic manipulation tools.


Subject(s)
Lysobacter/metabolism , Streptomyces/metabolism , Lipopeptides/metabolism , Polyketide Synthases/genetics , Multigene Family
2.
Article in English | WPRIM | ID: wpr-881041

ABSTRACT

Constitutively expression of the pathway-specific activators is an effective method to activate silent gene clusters and improve natural product production. In this study, nine shunt products of aminoansamycins (1-9) were identified from a recombinant mutant strain S35-LAL by overexpressed the large-ATP-binding regulator of the LuxR family (LAL) gene aas1 in Streptomyces sp. S35. All the compounds showed no anti-microbial, anti-T3SS and cytotoxic activities.


Subject(s)
Biological Products/metabolism , Lactams, Macrocyclic/metabolism , Multigene Family , Organisms, Genetically Modified , Streptomyces/metabolism
3.
Arq. Inst. Biol ; 87: e0142020, 2020. tab
Article in English | VETINDEX, LILACS | ID: biblio-1130108

ABSTRACT

The genus Streptomyces is associated with the ability to produce and excrete a variety of bioactive compounds, such as antibiotic, antifungal and antiviral. Biological active polyketide and peptide compounds with applications in medicine, agriculture and biochemical research are synthesized by PKS-I and NRPS genes. The evaluation of the presence of these genes associated with the biosynthesis of secondary metabolites in different phytopathogenic Streptomyces strains were performed using degenerated primers. The positive signal was observed in 58/63 Streptomyces strains for NRPS gene, 43/63 for PKS-I, and for PKS-II all the 63 strains showed positive signal of amplification. These strains also were tested with double layer agar-well technique against bacterial with clinical importance, and it was possible to observe the Streptomyces spp. strains were able to inhibit the growth of 14, 20, 13 and 3 isolates Gram-positive and Gram-negative bacteria, Staphylococcus aureus (ATCC 25923), Bacillus cereus (ATCC 14579), Pseudomonas aeruginosa (ATCC 27853) and Escherichia coli (ATCC 11775) respectively. The Streptomyces sp. strains IBSBF 2019 and IBSBF 2397 showed antibacterial activity against all four bacteria-target tested.(AU)


O gênero Streptomyces apresenta alta capacidade de produzir e excretar uma grande variedade de compostos biologicamente ativos, como antibióticos, antifúngicos e antivirais. Compostos biologicamente ativos de policetídeos e peptídeos com aplicações na medicina, agricultura e pesquisas bioquímicas são sintetizados pelos genes PKS-I e NRPS. A avaliação da presença desses genes associados à biossíntese de metabólitos secundários em diferentes linhagens de Streptomyces fitopatogênicas foi realizada através do uso de primers degenerados. O sinal positivo foi observado em 58/63 linhagens de Streptomyces para o gene NRPS, 43/63 para o gene PKS-I e, para o gene PKS-II, todas as 63 linhagens apesentaram o sinal positivo de amplificação. Essas linhagens também foram testadas através da técnica de dupla camada contra bactérias de importância clínica e foi possível observar que as linhagens de Streptomyces spp. foram capazes de inibir o crescimento de 14, 20, 13 e 3 isolados de bactérias Gram-positivas e Gram-negativas, Staphylococcus aureus (ATCC 25923), Bacillus cereus (ATCC 14579), Pseudomonas aeruginosa (ATCC 27853) e Escherichia coli (ATCC 11775), respectivamente. As linhagens de Streptomyces sp. ISBSF 2019 e 2397 apresentaram atividade antibacteriana contra todas as bactérias-alvo testadas.(AU)


Subject(s)
Pseudomonas aeruginosa/growth & development , Staphylococcus aureus/growth & development , Streptomyces/metabolism , Bacillus cereus/growth & development , Escherichia coli/growth & development , Anti-Bacterial Agents/metabolism , Peptide Synthases/genetics , Streptomyces/genetics , Gene Amplification , Polymerase Chain Reaction , Sequence Analysis, DNA , DNA Primers , Polyketide Synthases/genetics , Anti-Bacterial Agents/pharmacology
4.
Article in English | WPRIM | ID: wpr-1010506

ABSTRACT

Genome sequencing projects revealed massive cryptic gene clusters encoding the undiscovered secondary metabolites in Streptomyces. To investigate the metabolic products of silent gene clusters in Streptomyces chattanoogensis L10 (CGMCC 2644), we used site-directed mutagenesis to generate ten mutants with point mutations in the highly conserved region of rpsL (encoding the ribosomal protein S12) or rpoB (encoding the RNA polymerase β-subunit). Among them, L10/RpoB (H437Y) accumulated a dark pigment on a yeast extract-malt extract-glucose (YMG) plate. This was absent in the wild type. After further investigation, a novel angucycline antibiotic named anthrachamycin was isolated and determined using nuclear magnetic resonance (NMR) spectroscopic techniques. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis and electrophoretic mobility shift assay (EMSA) were performed to investigate the mechanism underlying the activation effect on the anthrachamycin biosynthetic gene cluster. This work indicated that the rpoB-specific missense H437Y mutation had activated anthrachamycin biosynthesis in S. chattanoogensis L10. This may be helpful in the investigation of the pleiotropic regulation system in Streptomyces.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Bacterial Proteins/genetics , Multigene Family , Mutagenesis, Site-Directed , Streptomyces/metabolism
5.
Braz. j. microbiol ; Braz. j. microbiol;49(4): 816-822, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974289

ABSTRACT

ABSTRACT Fifty seven soil-borne actinomycete strains were assessed for the antibiotic production. Two of the most active isolates, designed as Streptomyces ST-13 and DK-15 exhibited a broad range of antimicrobial activity and therefore they were selected for HPLC fractionation against the most suppressed bacteria Staphylococcus aureus (ST-13) and Chromobacterium violaceum (DK-15). LC/MS analysis of extracts showed the presence of polyketides factumycin (DK15) and tetrangomycin (ST13). The taxonomic position of the antibiotic-producing actinomycetes was determined using a polyphasic approach. Phenotypic characterization and 16S rRNA gene sequence analysis of the isolates matched those described for members of the genus Streptomyces. DK-15 strain exhibited the highest 16S rRNA gene sequence similarity to Streptomyces globosus DSM-40815 (T) and Streptomyces toxytricini DSM-40178 (T) and ST-13 strain to Streptomyces ederensis DSM-40741 (T) and Streptomyces phaeochromogenes DSM-40073 (T). For the proper identification, MALDI-TOF/MS profile of whole-cell proteins led to the identification of S. globosus DK-15 (accession number: KX527570) and S. ederensis ST13 (accession number: KX527568). To our knowledge, there is no report about the production of these antibiotics by S.globosus and S. ederensis, thus isolates DK15 and ST13 identified as S. globosus DK-15 and S.ederensis ST-13 can be considered as new sources of these unique antibacterial metabolites.


Subject(s)
Streptomyces/isolation & purification , Streptomyces/metabolism , Anti-Bacterial Agents/biosynthesis , Phylogeny , Pyridones/metabolism , Soil Microbiology , Streptomyces/classification , Streptomyces/genetics , Benz(a)Anthracenes/metabolism , DNA, Bacterial/genetics , Bacterial Typing Techniques
6.
Braz. j. microbiol ; Braz. j. microbiol;49(4): 731-741, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974291

ABSTRACT

ABSTRACT A bacterium isolated from Sterkfontein dam was confirmed to produce bioflocculant with excellent flocculation activity. The 16S rDNA nucleotide sequence analyses revealed the bacteria to have 99% similarity to Streptomyces platensis strain HBUM174787 and the sequence was deposited in the Genbank as Streptomyces platensis with accession number FJ 486385.1. Culture conditions for optimal production of the bioflocculant included glucose as a sole carbon source, resulting in flocculating activity of 90%. Other optimal conditions included: peptone as nitrogen source; presence of Mg2+ as cations and inoculum size of 1.0% (v/v) at neutral pH of 7. Optimum dose of the purified bioflocculant for the clarification of 4 g/L kaolin clay suspension at neutral pH was 0.2 mg/mL. Energy Dispersive X-ray analysis confirmed elemental composition of the purified bioflocculant in mass proportion (%w/w): carbon (21.41), oxygen (35.59), sulphur (26.16), nitrogen (0.62) and potassium (7.48). Fourier Transform Infrared Spectroscopy (FTIR) indicated the presence of hydroxyl, carboxyl, methoxyl and amino group in the bioflocculant. The bioflocculant produced by S. platensis removed chemical oxygen demand (COD) in river water and meat processing wastewater at efficiencies of 63.1 and 46.6% respectively and reduced their turbidity by 84.3 and 75.6% respectively. The high flocculating rate and removal efficiencies displayed by S. platensis suggests its industrial application in wastewater treatment.


Subject(s)
Streptomyces/chemistry , Bacterial Proteins/metabolism , Wastewater/chemistry , Streptomyces/isolation & purification , Streptomyces/genetics , Streptomyces/metabolism , Bacterial Proteins/genetics , Water Microbiology , Carbon/metabolism , Water Purification , Rivers/chemistry , Flocculation , Nitrogen/metabolism
7.
Braz. j. microbiol ; Braz. j. microbiol;49(4): 832-839, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974313

ABSTRACT

ABSTRACT Clavulanic acid is a β-lactam compound with potent inhibitory activity against β-lactamases. Studies have shown that certain amino acids play essential roles in CA biosynthesis. However, quantitative evaluations of the effects of these amino acids are still needed in order to improve CA production. Here, we report a study of the nutritional requirements of Streptomyces clavuligerus for CA production. Firstly, the influence of the primary nitrogen source and the salts composition was investigated. Subsequently, soybean protein isolate was supplemented with arginine (0.0-3.20 g L-1), threonine (0.0-1.44 g L-1), ornithine (0.0-4.08 g L-1), and glutamate (0.0-8.16 g L-1), according to a two-level central composite rotatable design. A medium containing ferrous sulfate yielded CA production of 437 mg L-1, while a formulation without this salt produced only 41 mg L-1 of CA. This substantial difference suggested that Fe2+ is important for CA biosynthesis. The experimental design showed that glutamate and ornithine negatively influenced CA production while arginine and threonine had no influence. The soybean protein isolate provided sufficient C5 precursor for CA biosynthesis, so that supplementation was unnecessary. Screening of medium components, together with experimental design tools, could be a valuable way of enhancing CA titers and reducing the process costs.


Subject(s)
Streptomyces/metabolism , Clavulanic Acid/biosynthesis , Culture Media/metabolism , Ornithine/analysis , Ornithine/metabolism , Streptomyces/genetics , Glutamic Acid/analysis , Glutamic Acid/metabolism , Culture Media/chemistry , Nitrogen/analysis , Nitrogen/metabolism
8.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;51(2): e6657, 2018. tab, graf
Article in English | LILACS | ID: biblio-889026

ABSTRACT

Surfactants are amphipathic compounds containing both hydrophilic and hydrophobic groups, capable to lower the surface or interfacial tension. Considering the advantages of the use of biosurfactants produced by microorganisms, the aim of this paper was to develop and characterize a biosurfactant produced by Streptomyces sp. DPUA1559 isolated from lichens of the Amazon region. The microorganism was cultured in a mineral medium containing 1% residual frying soybean oil as the carbon source. The kinetics of biosurfactant production was accompanied by reducing the surface tension of the culture medium from 60 to values around 27.14 mN/m, and by the emulsification index, which showed the efficiency of the biosurfactant as an emulsifier of hydrophobic compounds. The yield of the isolated biosurfactant was 1.74 g/L, in addition to the excellent capability of reducing the surface tension (25.34 mN/m), as observed from the central composite rotational design when the biosurfactant was produced at pH 8.5 at 28°C. The critical micelle concentration of the biosurfactant was determined as 0.01 g/mL. The biosurfactant showed thermal and pH stability regarding the surface tension reduction, and tolerance under high salt concentrations. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina, and to the seeds of lettuce (Lactuca sativa L.) and cabbage (Brassica oleracea L.). The biochemistry characterization of the biosurfactant showed a single protein band, an acid character and a molecular weight around 14.3 kDa, suggesting its glycoproteic nature. The results are promising for the industrial application of this new biosurfactant.


Subject(s)
Streptomyces/metabolism , Surface-Active Agents/metabolism , Lichens/microbiology , Reference Values , Seeds/drug effects , Temperature , Time Factors , Soybean Oil/chemistry , Colony Count, Microbial , Analysis of Variance , Spectroscopy, Fourier Transform Infrared , Culture Media , Electrophoresis, Polyacrylamide Gel , Fermentation , Hydrogen-Ion Concentration
9.
Electron. j. biotechnol ; Electron. j. biotechnol;28: 41-46, July. 2017. tab, ilus, graf
Article in English | LILACS | ID: biblio-1015839

ABSTRACT

Background: Streptomyces clavuligerus was the producer of clavulanic acid, claR, a pathway-specific transcriptional regulator in S. clavuligerus, positively regulates clavulanic acid biosynthesis. In this study, the promoter-less kanamycin resistance gene neo was fused with claR to obtain strain NEO from S. clavuligerus F613-1. The claR-neo fusion strain NEO was mutated using physical and chemical mutagens and then screened under high concentrations of kanamycin for high-yield producers of clavulanic acid. Results: The reporter gene neo was fused downstream of claR and used as an indicator for expression levels of claR in strain NEO. After three rounds of continuous treatment and screening, the high-yield clavulanic acid-producing strain M3-19 was obtained. In the shaking flask model, the clavulanic acid titer of M3-19 reached 4.33 g/L, which is an increase of 33% over the titer of 3.26 g/L for the starting strains S. clavuligerus F613-1 and NEO. Conclusions: Our results indicate that neo can be effectively used as a reporter for the expression of late-stage biosynthetic genes when screening for high-yield strains and that this approach has strong potential for improving Streptomyces strains of industrial value.


Subject(s)
Streptomyces/genetics , Streptomyces/metabolism , Kanamycin , Clavulanic Acid/biosynthesis , Transcription Factors/genetics , Transcription, Genetic , Biological Assay , Recombinant Proteins , Chromatography, High Pressure Liquid , Mutagenesis , Promoter Regions, Genetic , Genes, Reporter , Gene Fusion , Fermentation , Real-Time Polymerase Chain Reaction
10.
Electron. j. biotechnol ; Electron. j. biotechnol;19(4): 62-67, July 2016. ilus
Article in English | LILACS | ID: lil-793954

ABSTRACT

Background: A thermostable lipase MAS1 from marine Streptomyces sp. strain was considered as a potential biocatalyst for industrial application, but its production level was relatively low. Here, the effect of chaperones co-expression on the secretory expression of lipase MAS1 in Pichia pastoris was investigated. Result: Co-expression of protein disulfide isomerase (PDI), HAC1 and immunoglobulin binding protein could increase the expression level of lipase MAS1, whereas co-expression of Vitreoscilla hemoglobin showed a negative effect to the lipase MAS1 production. Among them, PDI co-expression increased lipase MAS1 expression level by 1.7-fold compared to the control strain harboring only the MAS1 gene. Furthermore, optimizing production of lipase MAS1 with Pichia pastoris strain X-33/MAS1-PDI in a 30-L bioreactor were conducted. Lower induction temperature was found to have a benefit effect for lipase MAS1 production. Lipase activity at 24 and 22°C showed 1.7 and 2.1-fold to that at 30°C, respectively. Among the induction pH tested, the highest lipase activity was obtained at pH 6.0 with activity of 440 U/mL after 144 h fermentation. Conclusion: Our work showed a good example for improving the production of recombinant enzymes in Pichia pastoris via chaperon co-expression and fermentation condition optimization.


Subject(s)
Streptomyces/metabolism , Lipase/biosynthesis , Pichia , Temperature , Cell Survival , Molecular Chaperones , Bioreactors , Protein Disulfide-Isomerases , Vitreoscilla , Fermentation , Batch Cell Culture Techniques , Hydrogen-Ion Concentration
11.
Braz. j. microbiol ; Braz. j. microbiol;46(4): 957-968, Oct.-Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769664

ABSTRACT

Abstract L-glutaminase was produced by Streptomyces canarius FR (KC460654) with an apparent molecular mass of 44 kDa. It has 17.9 purification fold with a final specific activity 132.2 U/mg proteins and 28% yield recovery. The purified L-glutaminase showed a maximal activity against L-glutamine when incubated at pH 8.0 at 40 °C for 30 min. It maintained its stability at wide range of pH from 5.0 11.0 and thermal stable up to 60 °C with Tm value 57.5 °C. It has high affinity and catalytic activity for L-glutamine (Km 0.129 mM, Vmax 2.02 U/mg/min), followed by L-asparagine and L-aspartic acid. In vivo, L-glutaminase showed no observed changes in liver; kidney functions; hematological parameters and slight effect on RBCs and level of platelets after 10 days of rabbit's injection. The anticancer activity of L-glutaminase was also tested against five types of human cancer cell lines using MTT assay in vitro. L-glutaminase has a significant efficiency against Hep-G2 cell (IC50, 6.8 μg/mL) and HeLa cells (IC50, 8.3 μg/mL), while the growth of MCF-7 cells was not affected. L-glutaminase has a moderate cytotoxic effect against HCT-116 cell (IC50, 64.7 μg/mL) and RAW 264.7 cell (IC50, 59.3 μg/mL).


Subject(s)
Animals/chemistry , Animals/drug effects , Animals/enzymology , Animals/metabolism , Animals/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/drug effects , Antineoplastic Agents/enzymology , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Biocatalysis/chemistry , Biocatalysis/drug effects , Biocatalysis/enzymology , Biocatalysis/metabolism , Biocatalysis/pharmacology , Cell Proliferation/chemistry , Cell Proliferation/drug effects , Cell Proliferation/enzymology , Cell Proliferation/metabolism , Cell Proliferation/pharmacology , Enzyme Stability/chemistry , Enzyme Stability/drug effects , Enzyme Stability/enzymology , Enzyme Stability/metabolism , Enzyme Stability/pharmacology , Glutaminase/chemistry , Glutaminase/drug effects , Glutaminase/enzymology , Glutaminase/metabolism , Glutaminase/pharmacology , Glutamine/chemistry , Glutamine/drug effects , Glutamine/enzymology , Glutamine/metabolism , Glutamine/pharmacology , HeLa Cells/chemistry , HeLa Cells/drug effects , HeLa Cells/enzymology , HeLa Cells/metabolism , HeLa Cells/pharmacology , /chemistry , /drug effects , /enzymology , /metabolism , /pharmacology , Humans/chemistry , Humans/drug effects , Humans/enzymology , Humans/metabolism , Humans/pharmacology , Kinetics/chemistry , Kinetics/drug effects , Kinetics/enzymology , Kinetics/metabolism , Kinetics/pharmacology , Streptomyces/chemistry , Streptomyces/drug effects , Streptomyces/enzymology , Streptomyces/metabolism , Streptomyces/pharmacology , Substrate Specificity/chemistry , Substrate Specificity/drug effects , Substrate Specificity/enzymology , Substrate Specificity/metabolism , Substrate Specificity/pharmacology
12.
Braz. j. microbiol ; Braz. j. microbiol;45(4): 1179-1186, Oct.-Dec. 2014. graf, tab
Article in English | LILACS | ID: lil-741267

ABSTRACT

Two mesophilic streptomycetes (S. violaceoruber and S. spiroverticillatus) were selected to study their Poly R-478 decolorization ability and lignocellulose solubilizing activity. Both strains were able to degrade Poly R-478 dye and ferulic acid during growth on a minimal salts medium. The Poly R-478 decolorizing activities of both strains were induced by adding different carbon sources to the culture media. S. violaceoruber could decolorize 63% of Poly R-478 after 24 h. Both strains could solubilize straw and produce acid-precipitable polymeric lignin (APPL) with different efficiency. From the major extracellular enzymes recovery of both strains on rice and wheat straw, we can predicate that the biodegradation process was partial indicating a possible utilization in biological delignification.


Subject(s)
Anthraquinones/metabolism , Lignin/metabolism , Polymers/metabolism , Streptomyces/metabolism , Biotransformation , Coumaric Acids/metabolism , Culture Media/chemistry , Oryza/metabolism , Plant Stems/metabolism , Streptomyces/growth & development , Triticum/metabolism
13.
Braz. j. microbiol ; Braz. j. microbiol;45(3): 919-928, July-Sept. 2014. graf, tab
Article in English | LILACS | ID: lil-727021

ABSTRACT

The selection of new microorganisms able to produce antimicrobial compounds is hoped for to reduce their production costs and the side effects caused by synthetic drugs. Clavulanic acid is a β-lactam antibiotic produced by submerged culture, which is widely used in medicine as a powerful inhibitor of β-lactamases, enzymes produced by bacteria resistant to antibiotics such penicillin and cephalosporin. The purpose of this work was to select the best clavulanic acid producer among strains of Streptomyces belonging to the Microorganism Collection of the Department of Antibiotics of the Federal University of Pernambuco (DAUFPE). Initially, the strains were studied for their capacity to inhibit the action of β-lactamases produced by Klebsiella aerogenes ATCC 15380. From these results, five strains were selected to investigate the batch kinetics of growth and clavulanic acid production in submerged culture carried out in flasks. The results were compared with the ones obtained by Streptomyces clavuligerus ATCC 27064 selected as a control strain. The best clavulanic acid producer was Streptomyces DAUFPE 3060, molecularly identified as Streptomyces variabilis, which increased the clavulanic acid production by 28% compared to the control strain. This work contributes to the enlargement of knowledge on new Streptomyces wild strains able to produce clavulanic acid by submerged culture.


Subject(s)
Clavulanic Acid/metabolism , Enzyme Inhibitors/metabolism , Streptomyces/isolation & purification , Streptomyces/metabolism , Enterobacter aerogenes/enzymology , Mass Screening , Streptomyces/growth & development , beta-Lactamases/metabolism
14.
Braz. j. microbiol ; Braz. j. microbiol;45(2): 743-751, Apr.-June 2014. ilus, tab
Article in English | LILACS | ID: lil-723113

ABSTRACT

The use of low cost agro-industrial residues for the production of industrial enzymes is one of the ways to reduce significantly production costs. Cellulase producing actinomycetes were isolated from soil and decayed agricultural wastes. Among them, a potential culture, strain NEAE-J, was selected and identified on the basis of morphological, cultural, physiological and chemotaxonomic properties, together with 16S rDNA sequence. It is proposed that strain NEAE-J should be included in the species Streptomyces albogriseolus as a representative of a novel sub-species, Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J and sequencing product was deposited in the GenBank database under accession number JN229412. This organism was tested for its ability to produce endoglucanase and release reducing sugars from agro-industrial residues as substrates. Sugarcane bagasse was the most suitable substrate for endoglucanase production. Effects of process variables, namely incubation time, temperature, initial pH and nitrogen source on production of endoglucanase by submerged fermentation using Streptomyces albogriseolus subsp. cellulolyticus have been studied. Accordingly optimum conditions have been determined. Incubation temperature of 30 ºC after 6 days, pH of 6.5, 1% sugarcane bagasse as carbon source and peptone as nitrogen source were found to be the optimum for endoglucanase production. Optimization of the process parameters resulted in about 2.6 fold increase in the endoglucanase activity. Therefore, Streptomyces albogriseolus subsp. cellulolyticus coud be potential microorganism for the intended application.


Subject(s)
Cellulase/isolation & purification , Cellulase/metabolism , Streptomyces/metabolism , Bacterial Typing Techniques , Cluster Analysis , Carbohydrates/analysis , Cellulose/metabolism , Culture Media/chemistry , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Hydrogen-Ion Concentration , Molecular Sequence Data , Phylogeny , /genetics , Sequence Analysis, DNA , Saccharum/metabolism , Streptomyces/classification , Streptomyces/growth & development , Streptomyces/isolation & purification , Temperature , Time Factors
15.
Article in English | IMSEAR | ID: sea-163101

ABSTRACT

Aims: To optimize the process parameters for enhanced production of bioactive metabolites by Streptomyces tritolerans DAS 165T. Place and Duration of Study: Department of Botany and Microbiology, April 2012 to August 2012. Methodology: Agar well diffusion assay was employed to study the effect of environmental parameters such as incubation period, pH, temperature and salt concentration and influence of various nutrients such as carbon and nitrogen sources and minerals on the bioactive metabolite production by Streptomyces tritolerans DAS 165T.  Results: The production of antimicrobial metabolite was high when the strain was cultured for six days at 35ºC in medium (pH 7.5) with sucrose at the concentration of 2% (carbon source), soya peptone at the concentration of 1% (nitrogen source) and sodium chloride at the concentration of 5%.  Conclusion: This is the first report on the optimization of bioactive metabolite production by Streptomyces tritolerans DAS 165T. As the strain exhibited potent antimicrobial activity, it may be explored for biotechnological purposes.


Subject(s)
Biological Products/biosynthesis , Biological Products/metabolism , Environment , Microbial Sensitivity Tests , Microbial Viability , Nutritional Status , Streptomyces/classification , Streptomyces/metabolism , Streptomyces/physiology
16.
Braz. j. microbiol ; Braz. j. microbiol;45(1): 303-312, 2014. ilus, tab
Article in English | LILACS | ID: lil-709457

ABSTRACT

A highly potent secondary metabolite producing actinomycetes strain is isolated from marine soil sediments of Visakhapatnam sea coast, Bay of Bengal. Over all ten strains are isolated from the collected soil sediments. Among the ten actinomycetes strains the broad spectrum strain RSPSN2 was selected for molecular characterization, antibiotic production and its purification. The nucleotide sequence of the 1 rRNA gene (1261 base pairs) of the most potent strain evidenced a 96% similarity with Streptomyces parvulus 1044 strain, Streptomyces parvulus NBRC 13193 and Streptomyces parvulus BY-F. From the taxonomic features, the actinomycetes isolate RSPSN2 matches with Streptomyces parvulus in the morphological, physiological and biochemical characters. Thus, it was given the suggested name Streptomyces parvulus RSPSN2. The active metabolite was extracted using ethyl acetate (1:3, v/v) at pH 7.0. The separation of active ingredient and its purification was performed by using both thin layer chromatography (TLC) and column chromatography (CC) techniques. Spectrometric studies such as UV-visible, FTIR, and NMR and mass were performed. The antibacterial activity of pure compound was performed by cup plate method against some pathogenic bacteria including of streptomycin resistant bacteria like (Pseudomonas mirabilis. Pseudomonas putida and Bacillus cereus). In conclusion, the collected data emphasized the fact that a polypeptide antibiotic (Actinomycin D) was produced by Streptomyces parvulus RSPSN2.


Subject(s)
Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Peptides/isolation & purification , Peptides/pharmacology , Streptomyces/metabolism , Anti-Bacterial Agents/chemistry , Bacterial Typing Techniques , Chromatography, Liquid , Chromatography, Thin Layer , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Geologic Sediments/microbiology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , India , Magnetic Resonance Spectroscopy , Microbial Sensitivity Tests , Phylogeny , Peptides/chemistry , RNA, Ribosomal/genetics , Sequence Analysis, DNA , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared , Streptomyces/isolation & purification
17.
Braz. j. microbiol ; Braz. j. microbiol;44(4): 1049-1057, Oct.-Dec. 2013. ilus, graf, tab
Article in English | LILACS | ID: lil-705270

ABSTRACT

Clavulanic acid (CA) is a powerful inhibitor of the beta-lactamases, enzymes produced by bacteria resistants to penicillin and cefalosporin. This molecule is produced industrially by strains of Streptomyces clavuligerus in complex media which carbon and nitrogen resources are supplied by inexpensive compounds still providing high productivity. The genetic production improvement using physical and chemical mutagenic agents is an important strategy in programs of industrial production development of bioactive metabolites. However, parental strains are susceptible to loss of their original productivity due genetic instability phenomenona. In this work, some S. clavuligerus mutant strains obtained by treatment with UV light and with MMS are compared with the wild type (Streptomyces clavuligerus ATCC 27064). The results indicated that the random mutations originated some strains with different phenotypes, most divergent demonstrated by the mutants strains named AC116, MMS 150 and MMS 54, that exhibited lack of pigmentation in their mature spores. Also, the strain MMS 150 presented a larger production of CA when cultivated in semi-synthetics media. Using other media, the wild type strain obtained a larger CA production. Besides, using the modifed complex media the MMS 150 strain showed changes in its lipolitic activity and a larger production of CA. The studies also allowed finding the best conditions for a lipase activity exhibited by wild type S. clavuligerus and the MMS150 mutant.


Subject(s)
Clavulanic Acid/metabolism , Metabolic Engineering , Mutagenesis , Mutation , Streptomyces/metabolism , Culture Media/chemistry , Lipase/metabolism , Methyl Methanesulfonate , Streptomyces/drug effects , Streptomyces/genetics , Streptomyces/radiation effects , Ultraviolet Rays
18.
Braz. j. infect. dis ; Braz. j. infect. dis;16(5): 466-471, Sept.-Oct. 2012. ilus, tab
Article in English | LILACS | ID: lil-653436

ABSTRACT

Streptomyces is a genus of Gram-positive bacteria that grows in various environments, and its shape resembles filamentous fungi. The morphological differentiation of Streptomyces involves the formation of a layer of hyphae that can differentiate into a chain of spores. The most interesting property of Streptomyces is the ability to produce bioactive secondary metabolites, such as antifungals, antivirals, antitumorals, anti-hypertensives, immunosuppressants, and especially antibiotics. The production of most antibiotics is species specific, and these secondary metabolites are important for Streptomyces species in order to compete with other microorganisms that come in contact, even within the same genre. Despite the success of the discovery of antibiotics, and advances in the techniques of their production, infectious diseases still remain the second leading cause of death worldwide, and bacterial infections cause approximately 17 million deaths annually, affecting mainly children and the elderly. Self-medication and overuse of antibiotics is another important factor that contributes to resistance, reducing the lifetime of the antibiotic, thus causing the constant need for research and development of new antibiotics.


Subject(s)
Humans , Anti-Bacterial Agents/biosynthesis , Streptomyces/metabolism , Anti-Bacterial Agents/pharmacology , Streptomyces/physiology
19.
Electron. j. biotechnol ; Electron. j. biotechnol;15(4): 6-6, July 2012. ilus, tab
Article in English | LILACS | ID: lil-646956

ABSTRACT

Response surface methodology was used to optimize the fermentation conditions for the production of pristinamycin by immobilization of Streptomyces pristinaespiralis F213 in shaking flask cultivation. Seed medium volume, fermentation medium volume and shaking speed of seed culture were found to have significant effects on pristinamycin production by the Plackett-Burman design. The steepest ascent method was adopted to approach the vicinity of optimum space, followed by central composite design for further optimization. A quadratic model was built to fit the pristinamycin production. The optimum conditions were found to be seed medium volume of 29.5 ml, fermentation medium volume of 28.8 ml, and shaking speed of seed culture at 204 rpm. At the optimum conditions, a production of 213 mg/l was obtained, which was in agreement with the maximum predicted pristinamycin yield of 209 mg/l. This is the first report on pristinamycins production by immobilized S. pristinaespiralis using response surface methodology.


Subject(s)
Fermentation , Pristinamycin/biosynthesis , Streptomyces/metabolism , Culture Techniques
20.
Indian J Biochem Biophys ; 2011 Oct; 48(5): 331-335
Article in English | IMSEAR | ID: sea-135336

ABSTRACT

Biosynthesis of gold nanoparticles by Streptomycetes from Himalayan Mountain was undertaken for the first time. Out of 10 actinomycete strains tested, four strains (D10, HM10, ANS2 and MSU) showed evidence for the intracellular biosynthesis of gold nanoparticles, among which the strain HM10 showed high potency. Presence of spherical and rod shaped gold nanoparticles in mycelium of the strain HM10 was determined by transmission electron microscopy (TEM) and X-ray diffraction analysis. The average particle size ranged from 18-20 nm. UV spectral analysis indicated that the reduction of chloroauric acid (HAuCl4) occurred within 24 h of reaction period. Further, the strain HM10 showed enhanced growth at 1 and 10 mM concentration of HAuCl4. The gold nanoparticles synthesized by the strain HM10 showed good antibacterial activity against S. aureus and E. coli in well-diffusion method. The potential actinomycete HM10 strain was phenotypically characterized and identified as Streptomyces viridogens (HM10). Thus, actinomycete strain HM10 reported in this study is a newly added source for the biosynthesis of gold nanoparticles.


Subject(s)
Actinobacteria/metabolism , Anti-Bacterial Agents/biosynthesis , Anti-Bacterial Agents/chemistry , Chlorides/chemistry , Chlorides/metabolism , Chlorides/pharmacology , Escherichia coli/drug effects , Gold Compounds/chemistry , Gold Compounds/metabolism , Gold Compounds/pharmacology , Microscopy, Electron, Transmission/methods , Nanoparticles/chemistry , Nanotechnology/methods , Staphylococcus aureus/drug effects , Streptomyces/metabolism , X-Ray Diffraction
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