Bioflocculant production from Streptomyces platensis and its potential for river and waste water treatment

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.

Secondary Metabolites from Streptomyces sp CSDX076 for the First Time / Metabolitos secundarios de Streptomyces SP CSDX076 por primera vez

ABSTRACT Objective: To investigate the secondary metabolites from the cultures of Streptomyces sp CSDX076. Methods: The compounds were isolated using column chromatography and RP-18 medium-pressure liquid chromatography. Their structures were elucidated by one-dimensional and two-dimensional nuclear magnetic resonance spectroscopic methods in combination with mass spectrometry experiments. Results: Four compounds were isolated from the cultures of Streptomyces sp CSDX076 and identified as aurantiamide benzoate, deoxytryptoquivaline, 2-acetyl-3,5-dihydroxyl-benzene acetic acid, and 2-acetyl-3,5-dihydroxyl-benzene ester. Conclusion: It was the first time that the four isolated compounds were obtained from the Streptomyces genus.

RESUMEN Objetivo: Investigar los metabolitos secundarios de los cultivos de Streptomyces sp CSDX076. Métodos: Los compuestos fueron aislados usando la cromatografía de columna y cromatografía líquida RP-18 de presión media. Sus estructuras fueron dilucidadas mediante métodos espectroscópicos de resonancia magnética nuclear unidimensional y bidimensional, combinados con experimentos de espectrometría de masa. Resultados: Cuatro compuestos de culturas de Streptomyces sp CSDX076 fueron aislados e identificados como benzoato de aurantiamida, deoxitriptoquivalina, ácido acético 2-acetil-3,5-dihidroxil-benzeno, y éster 2-acetil-3,5-dihidroxil-benzeno. Conclusión: Fue la primera vez que los cuatro compuestos aislados se obtuvieron del género Streptomyces.

Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens / Seleção, produção e caracterização bioquímica de uma nova enzima fibrinolítica produzida por Streptomyces sp. (Streptomycetaceae) isolada de liquens da Amazônia

Thrombosis is a pathophysiological disorder caused by accumulation of fibrin in the blood. Fibrinolytic proteases with potent thrombolytic activity have been produced by diverse microbial sources. Considering the microbial biodiversity of the Amazon region, this study aimed at the screening, production and biochemical characterization of a fibrinolytic enzyme produced by Streptomyces sp. isolated from Amazonian lichens. The strain Streptomyces DPUA1576 showed the highest fibrinolytic activity, which was 283 mm2. Three variables at two levels were used to assess their effects on the fibrinolytic production. The parameters studied were agitation (0.28 - 1.12 g), temperature (28 - 36 ºC) and pH (6.0 - 8.0); all of them had significant effects on the fibrinolytic production. The maximum fibrinolytic activity (304 mm2) was observed at 1.12 g, 28 ºC, and pH of 8.0. The crude extract of the fermentation broth was used to assess the biochemical properties of the enzyme. Protease and fibrinolytic activities were stable during 6 h, at a pH ranging from 6.8 to 8.4 and 5.8 to 9.2, respectively. Optimum temperature for protease activity ranged between 35 and 55 °C, while the highest fibrinolytic activity was observed at 45 ºC. Proteolytic activity was inhibited by Cu2+ and Co2+ ions, phenylmethylsulfonyl fluoride (PMSF) and pepstatin A, which suggests that the enzyme is a serine protease. Enzymatic extract cleaved fibrinogen at the subunits Aalpha-chain, Abeta-chain, and gama-chain. The results indicated that Streptomyces sp. DPUA 1576 produces enzymes with fibrinolytic and fibrinogenolytic activity, enzymes with an important application in the pharmaceutical industry.

A trombose é uma doença patofisiológica causada pelo acúmulo de fibrina no sangue. Proteases fibrinolíticas com potente atividade trombolítica são produzidas por diversas fontes microbianas. Considerando a biodiversidade microbiana da região amazônica, o presente estudo teve como objetivo a seleção, produção e caracterização bioquímica da enzima fibrinolítica de Streptomyces sp. isolado de líquens da Amazônia. Streptomyces DPUA1576 foi a melhor produtora com atividade fibrinolítica de 283 mm2. Três variáveis em dois níveis foram utilizadas para determinar as variáveis mais relevantes na produção da enzima fibrinolítica (FA). Os parâmetros estudados foram agitação (0.28 - 1.12 g), temperatura (28 - 36 ºC) e pH (6.0 - 8.0) e todos obtiveram efeitos significativos na produção fibrinolítica. A maior atividade fibrinolítica (304 mm2) foi obtida a 1.12 g, 28 ºC e pH 8.0. O extrato bruto da fermentação foi usado para determinar as propriedades bioquímicas da enzima. Atividades proteásica e fibrinolítica foram estáveis durante 6 horas no intervalo de pH entre 6.8 - 8.4 e 5.8 - 9.2, respectivamente. Temperatura ótima para a atividade proteásica foi entre 35 - 55 °C, enquanto que para a atividade fibrinolítica foi de 45 ºC. Atividade proteásica foi inibida por íons Cu2+ e Co2+, fluoreto de fenilmetilsulfonil e pepstatina A, na qual sugere que a enzima é uma serino-protease. O extrato enzimático degradou o fibrinogênio nas subunidades Aalfa, Abeta e gama . Os resultados apresentados indicam que Streptomyces sp. DPUA 1576 produz enzimas com atividade fibrinolítica e fibrinogenolítica, enzimas com aplicações importantes na indústria farmacêutica.

Influence of glucose and stirring in the fermentation process in order to produce anti-Candida metabolites produced by Streptomyces sp

ABSTRACT This study evaluated the influence of glucose and stirring in the fermentation process in order to produce anti-Candida metabolites produced by Streptomyces sp. MPO4 isolated from Amazon soil. The anti-Candida metabolites production was registered after 24 h of fermentation in stirred ISP2 medium, having antifungal inhibition halos between 12.3 mm and 25.3 mm, yielding higher production of anti-Candida agents after 96 h. Stirring was a determining factor for the production of anti-Candida secondary metabolites, since the absence of glucose reflected in the late production of the antifungal starting from Streptomyces sp.

RESUMO Este estudo avaliou a influência da glicose e agitação no processo de fermentação para a produção de metabólitos anti-Candida produzidos por Streptomyces sp. MPO4 isolado do solo da Amazônia. A produção dos metabólitos anti-Candida foi registrada a partir de 24 h de fermentação sob agitação em meio ISP2, apresentando halos de inibição entre 12,3 mm e 25,3 mm, obtendo-se maior produção do antifúngico em 96 h. A agitação foi um fator determinante para a produção de metabólitos secundários anti-Candida e a ausência de glicose refletiu na produção tardia do antifúngico a partir do Streptomyces sp.

Kinetic properties of Streptomyces canarius L- Glutaminase and its anticancer efficiency

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).

Antitumor compounds from Streptomyces sp. KML-2, isolated from Khewra salt mines, Pakistan

BACKGROUND: Actinomycetes are gram positive bacteria with high G + C content in their DNA and are capable of producing variety of secondary metabolites. Many of these metabolites possess different biological activities and have the potential to be developed as therapeutic agents. The aim of the present study was to screen actinomycetes inhabiting halophilic environment such as Khewra salt mines present in Pakistan for cytotoxic and antitumor compounds. RESULTS: An actiomycetes strain designated as Streptomyces sp. KML-2 was isolated from a saline soil of Khewra salt mines, Pakistan. The strain Streptomyces sp. KML-2 showed 84 % cytotoxic activity against larvae of Artemiasalina. In the screening phase, the strain exhibited significant antitumor activity with IC50 values of 12, 48 and 56 µg/ml against Hela, MDBK and Vero cell lines, respectively. After that extract from 20 l fermentation was used to purify secondary metabolites by several chromatographic techniques. Structure elucidation of isolated compounds revealed that it is highly stable producer of Chromomycin SA (1) and 1-(1H-indol-3-yl)-propane-1,2,3-triol (2). Both of the isolated compounds showed significant antitumor activity against Hela and MCF-7 cancer cell lines (IC50 values 8.9 and 7.8 µg/ml against Hela; 12.6 and 0.97 µg/ml against MCF-7, respectively). The 16S rRNA gene sequence (1437 bp) of the strain confirm its identity (99 %) with Streptomyces griseus. CONCLUSIONS: From this research work we were successful in isolating two potent antitumor compounds, Chromomycin SA and 1-(1H-indol-3-yl)-propane-1,2,3-triol from Streptomyces KML-2 strain, isolated from Khewra salt mine. As such this is the second report which confirms that S. griseus can produce Chromomycin SA without introducing any mutagenesis in its biosynthesizing gene cluster and isolated indole derivative is being reported first time from any member of actinomycetes group with having novel antitumor activity against Hela and MCF-7 cells Nucleotide sequences: Nucleotide sequence data reported are available in the GenBank database under the accession number: GenBank KJ009562.

Anti-fungal potentials of extracellular metabolites of Western Ghats isolated Streptomyces sp. NII 1006 against moulds and yeasts.

Realization of hazardious effects of chemical fungicides has led to an interest in the usage of biocontrol agents. The present study, therefore, evaluates the biocontrol efficacy of Western Ghats (India) soil bacterial isolates. A potential strain NII 1006 was evaluated for its antagonistic property against a diverse range of moulds and yeasts. The strain was characterized morphologically, biochemically and molecularly, which revealed the isolate belonged to Streptomyces genus. Organic solvent extracts of NII 1006 culture filtrates inhibited the growth of the test pathogens indicating that growth suppression was due to extracellular anti-fungal metabolites present in the culture filtrates. The strain produced extracellular chitinase enzyme in addition to some stable partially purified anti-fungal compounds. Morphological changes such as hyphae degradation into debris and abnormal shapes were observed in test fungi and yeast grown on potato dextrose broth that contained the NII 1006 culture filtrate. The cell free supernatant has a tolerance to wide range of pH, temperature and enzymes such as lipase and protease. The biocontrol potential of NII 1006 strain may be correlated significantly with their ability to produce antibiotics as well as extracellular hydrolytic enzymes particularly chitinolytic enzyme.

Isolation, characterization and antifungal activity of Streptomyces sampsonii GS 1322.

For new antifungal antibiotics from actinomycetes, a strain of Streptomyces GS 1322 was isolated from a sample of garden soil. The strain was found to possess antagonistic activity against four fungi i.e., Candida albicans, Aspergillus niger, Microsporum gypseum and Trichophyton sp. The strain was identified as Streptomyces sampsonii and the antifungal compound produced by it was found to be the heptaene group of polyene antibiotics.

A non-polyene antifungal antibiotic from Streptomyces albidoflavus PU 23.

In all 312 actinomycete strains were isolated from water and soil samples from different regions. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising strain, Streptomyces albidoflavus PU 23 with strong antifungal activity against pathogenic fungi was selected for further studies. Antibiotic was extracted and purified from the isolate. Aspergillus spp. was most sensitive to the antibiotic followed by other molds and yeasts. The antibiotic was stable at different temperatures and pH tested and there was no significant loss of the antifungal activity after treatment with various detergents and enzymes. Synergistic effect was observed when the antibiotic was used in combination with hamycin. The antibiotic was fairly stable for a period of 12 months at 4 degree C. The mode of action of the antibiotic seems to be by binding to the ergosterol present in the fungal cell membrane resulting in the leakage of intracellular material and eventually death of the cell. The structure of the antibiotic was determined by elemental analysis and by ultraviolet (UV), Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR) and liquid chromatography mass spectra (LCMS). The antibiotic was found to be a straight chain polyhydroxy, polyether, non-proteinic compound with a single double bond, indicating a nonpolyene antifungal antibiotic.

JU-2, a novel phosphorous-containing antifungal antibiotic from Streptomyces kanamyceticus M8.

A novel phosphorous-containing antifungal antibiotic JU-2 was isolated from Streptomyces kanamyceticus M8. Quantitative chemical analysis shows the presence of two phenylalanines, two glucose, one linoleic acid, one crucic acid and one phosphonamide moiety per molcule of the antibiotic. JU-2 shows strong inhibitory activity against various pathogenic and non-pathogenic fungi but no activity against bacteria and yeast.

Screening and identification of antibiotic producing strains of Streptomyces.

About 450 actinomycetes were isolated from nearly 100 soil samples collected from different parts of West Bengal. The isolates were screened on the basis of their inhibitory effect against test organisms. Finally two potent antibiotic producers were chosen having maximum inhibitory effect on both gram positive and gram negative test bacteria. On the basis of morphological, structural, physiological and biochemical characters, the two potent antibiotic producers were identified as Streptomyces violaceus-niger and S. antibioticus.