Streptomyces Violascens Research Articles

Effects of Four Strains of Actinomycetes on the Content of Terpenoids in Baijiu.

Terpenoids not only are an important health factor in baijiu but also contribute to the elegance and finesse of baijiu, and actinomycetes act as an important source of terpenoids in baijiu. Four strains of actinomycetes-Streptomyces violascens (SPQ1), S. sampsonii (SPS1), S. thermophilus (SPG1), and S. griseus (SPH1)-obtained from the Daqu, pit mud, fermented grains and air, respectively, in the production of baijiu were used in solid-state and liquid fermentation with five brewing raw materials as the substrates. The terpenoids in the metabolites were analyzed and compared using gas chromatography-mass spectrometry (GC-MS). We found that the four strains of actinomycetes produced 31 terpenoids from the hydrolysates of five fermentation substrates during liquid fermentation, and the total terpenoid content was 989.94 μg/kg in the fermentation products. After 28 days of solid-state fermentation, the four actinomycete strains produced 64 terpenoids using the five fermentation substrates, and the total terpenoid content was 23,651.52 μg/kg in the fermentation products. The different fermentation substrates and fermentation methods have a great influence on the terpenoids produced by actinomycetes.

Synergistic Activity of Cyclo(phe-pro) Antibiotic from Streptomyces violascens VS in Reducing the Drug Resistance Burden of Clinically Significant Pathogens

A potential novel antibiotic-producing soil Streptomyces violascens VS was isolated and confirmed through 16S rDNA sequencing. A novel antibiotic from S. violascens VS was extracted from the mass-cultured broth through organic solvents using chloroform: methanol by column and thin layer chromatography. Among the ten fractions, collected from elution by 9:3, 1:1, and 7:3 chloroform: methanol mixtures, the` 3rd fraction (after 3 minutes) of the 7:3 mixture showed maximum bactericidal activity against P. aeruginosa, E. coli, Bacillus sp., K. pneumoniae, and Proteus sp., with zones of inhibition of 18, 12, 19, 18, and 14 mm respectively. The equivalent fraction yielded 2.3 g/l of pure antibiotic. In the GC-MS spectrum the active compound identified as Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(phenylmethyl), or Cyclo(phe-pro). The antibiotic’s fractional inhibitory concentration index (FICI) showed synergistic and partially synergistic activity with ten different commercial antibiotics such as ampicillin, sulfamethoxazole, erythromycin, amoxicillin, azithromycin, ofloxacin, levofloxacin, cefadroxil, cefixime, and ciprofloxacin against Gram-positive and Gram-negative bacteria, especially with cell wall synthesis inhibiting antibiotics. Thus, the synergistic usage of Cyclo(phe-pro) peptide antibiotic with commercial antibiotics recognized to have potential broad spectrum activity and possibility to control infections caused by MDR strains.

Characterisation of Streptomyces violascens OC125-8 lipase for oily wastewater treatment.

In this study, the lipase-producing bacterium Streptomyces violascens (GenBank number MF621564) was identified, and the extracellular S. violascens OC125-8 lipase produced by this strain was characterised for use in wastewater treatment. The lipase was partially purified by ammonium sulphate precipitation at a final yield of 3.28-fold purification and a recovery of 56%. The S. violascens OC125-8 lipase exhibited optimum catalytic activity at 40°C and pH 8.0; it was stable at 30-40°C with more than 86% residual activity after 1h; it was also stable over a relatively broad pH range of pH 7.0-11.0, retaining 83.3-100% activity. V max and K m values were calculated as 0.61µmol/min/mg and 0.259mM, respectively. Enzyme activity significantly increased in the presence of Fe2+ ion but was inhibited by Ca2+, Mn2+, Cu2+ and Mg2+. The addition of a serine protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), strongly inhibited enzyme activity while ethylenediaminetetraacetic acid (EDTA), a metal chelating agent, had no inhibitory effect. The enzyme was fairly stable in the presence of surfactants as well as sodium perborate. Examination of commercial detergent tolerance revealed that the lipase was strongly stable in Tursil (88%), Pril (97%) and Fairy (98.5%), while the lipase was activated in Omo (113.4%) and Ariel (128.3%). Moreover, the lipase showed highest activity towards olive oil (100%), sunflower oil (90%) and burned sunflower oil (55%), while corn oil (44%) and burned olive oil (15%) were less hydrolysed by the enzyme. These properties demonstrate that S. violascens OC125-8 lipase is an ideal choice for oily wastewater management.

Biological control of toxigenic citrus and papaya-rotting fungi by Streptomyces violascens MT7 and its extracellular metabolites.

An Indian indigenous, Loktak Lake soil isolate Streptomyces violascens MT7 was assessed for its biocontrol potential both in vitro and in vivo against toxigenic fruit-rotting fungi. Strain MT7 exhibited broad-spectrum antifungal activity against various pathogenic postharvest fungi of citrus and papaya. In shake-flask fermentation, antagonist S. violascens MT7 highly produced extracellular antifungal metabolites in early stationary growth phase in glucose-yeast extract-malt extract (M93) broth. Both extracellular culture fluid (ECF) and its n-butanol extract showed significant broad-spectrum fungal mycelial inhibition of several tested fruit-rotting fungi. Antifungal metabolite was found to be heat stable, nonpeptidic, and polyene type antibiotic. The lowest minimum inhibitory concentration (MIC) of n-butanol extract against Colletotrichum gloeosporioides MTCC 9664 and Aspergillus niger MTCC 281 was 0.0312 and 0.0625 mg/ml, respectively. Purification of n-butanol extract through silica gel chromatography resulted in partial purification of bioactive metabolite and the TLC autobiography revealed the presence of single antifungal metabolite with Rf value of 0.755. In vivo bioassays demonstrated the biocontrol potential of tested biocontrol agents on fruit-rotting fungi. Use of cell suspension of S. violascens MT7, extracellular metabolite(s), and n-butanol extract significantly (p < 0.05) reduced sour-rot development on Citrus reticulata Blanco (oranges) and soft-rot development on papaya fruits. Therefore, these results strongly suggest a high potential for application of S. violascens MT7 and its extracellular metabolites as an effective eco-friendly alternative to synthetic fungicides for controlling toxigenic citrus and papaya-rotting fungi.

Xylanase Activity of Streptomyces violascences BF 3.10 on Xylan Corncobs and its Xylooligosaccharide Production

Corn is one of the important carbohydrate sources in Indonesia that is mainly used for food and industrial materials. In addition, the byproducts of corn, such as corncobs, have been reported as xylan-containing materials that can be utilized as substrate in xylooligosaccharides (XOS) production. XOS are natural prebiotic fibers that can enhance the performance of animal’s digestive system. The main objective of this study was to exploit xylan from corncobs to produce XOS. The research consisted of extraction and production of xylan from corncobs and the synthesis of XOS from corncob-produced xylan. The corncob and Streptomyces violascens BF 3.10 xylanase is a collection of PPSHB IPB Laboratory. Corncobs xylan extracted by using alkaline method and reducting sugar was analyzed by dinitrosalicylic acid method. The xylan extraction from corncobs could produce 7.93% (w/w) of xylan. The activity of S. violascens BF 3.10 xylanase on the substrate of concorb-produced xylan was 6.4 U/mL at the optimum temperature of 60 °C in 50 mM phosphate buffer with pH 5.5. The thin layer chromatography analysis indicated that 1% (w/v) corn-cob xylan could produce XOS with degree of polymerization (DP) 3.92. XOS, with DP ranging from 2-4, could be used as a livestock feed mixture to stimulate the growth of normal microbes in the gastrointestinal tract of livestock. Key words: corncobs, Streptomyces violascens BF 3.10, xylan, xylanase

Enzymatic Hydrolysis of Porang by Streptomyces violascens BF 3.10 Mannanase for the Production of Mannooligosaccharides

Porang ( Amorphophallus muelleri Blume) is an indigenous Indonesian plant containing high hemicellulose as a source of glucomannan. An alternative way to produce a good quality of mannooligosaccharides was through hydrolysis of glucomannan by endo-β mannnase from actynomicetes. Based on 16S rRNA analysis, BF 3.10 isolate, isolated from Bukit Duabelas National Park soil, Jambi was identified as Streptomyces violascens BF 3.10 . Reducing sugar was analyzed by dinitrosalicylic acid methods. The highest reducing sugar was achieved at the 72 hours of incubation. Mannanase of isolate BF 3.10 had the highest activity at pH 6 and temperature of 70 °C with enzyme activity of 16.38 U/mL and was stable at 4 °C for 48 h. During 5-hour of hydrolysis with substrate concentration of 0.25%, 0.5%, and 1% porang glucomannan dissolved in 10 mL enzyme, mannooligosaccharides were produced with the degree of polymerization of 2-3. Visualization of the products by using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) methods showed that mannooligosaccharides produced comprised of glucose, mannobiose, mannotriose, and mannotetraose. The degree of polymerization and the simple sugars produced indicated that mannanase produced by S. violascens actively catalyzed the hydrolysis of 1.4-β-D-mannoside linkage from β-1.4-mannan backbone, that eventually produced simple sugars of mannooligosaccharides. Key words: glucomannan, mannanase, mannooligosaccharides, porang, Streptomyces violascens

Isolation, Purification and Partial Characterization of an Antifungal Agent Produced by Salt-tolerant Alkaliphilic Streptomyces violascens IN2-10

An actinobacterium strain designated as IN2-10 was isolated from Garhwal region. IN2-10 was subjected to polyphasic characterization. The organism produced white to cream colored aerial mycelium with brown diffusible pigment. This isolate showed growth at 11 % NaCl, pH 4–10, and a temperature range of 15–37 °C. The isolate IN2-10 was found to belong to the genus Streptomyces and it was closely related to Streptomyces violascens (98.57 %, 16S rDNA). Of the various media tested, the high yield of active metabolite for isolate was observed in glucose soyabean meal medium at pH 10, with salt concentration of 3 % (w/v) and at a temperature of 37 °C. The broad spectrum antifungal activity of metabolite was observed against yeasts (Candida albicans, C. parapsilosis, and C. tropicalis) as well as molds (Microsporum gypseum, M. canis, Trichophtyton rubrum, Aspergillus flavus, A. fumigatus, and A. niger). The active metabolite produced by the strain was partially characterized and was found to belong to the heptaene group of polyene class of antifungal agents. In view of the decline in the discovery of new lead compounds in recent years, further investigations would require to lead to some useful products.

Synergistic effects of a cellulase-producing Micromonospora carbonacea and an antibiotic-producing Streptomyces violascens on the suppression of Phytophthora cinnamomi root rot of Banksia grandis

Three polyvalent Streptomyces phages were used to isolate four Micromonospora species (M. carbonacea, M. chalcea, M. purpureochromogenes, and M. inositola) from mine-site rhizosphere soils in Western Australia. Streptomyces violascens was isolated using selective isolation techniques from the same soils. The Micromonspora spp. were examined for their ability to produce cellulases. Micromonospora carbonacea, M. chalcea, and M. purpureochromogenes, which were found to produce the enzyme, caused lysis of Phytophthora cinnamomi hyphae. Glasshouse trials showed that the use of the cellulase-producing M. carbonacea isolate, in conjunction with the antibiotic-producing S. violascens isolate, had a synergistic effect on the suppression of the Phytophthora root rot and in promoting growth of Banksia grandis. The importance of using a number of antagonists with different antagonistic abilities to control plant pathogenic fungi is discussed. Keywords: biological control, Micromonospora carbonacea, Streptomyces violascens, cellulases, Phytophthora cinnamomi.

Some Enzymatic Properties of 3β-Hydroxysteroid Oxidase Produced by Streptomyces violascens

The 3beta-hydroxysteroid oxidase produced by Streptomyces violascens was purified from the culture broth by procedures including batch-wise treatment on DEAE-cellulose, ammonium sulfate fraction, gel filtration on Sephadex G-75, and column chromatography on DEAE-cellulose. The highly purified enzyme preparation exhibited no significant absorption maxima in the visible region other than a maximum at 280 nm. Optimum pH and temperature for the enzyme activity were approximately pH 7.5 and 50 degree, respectively. The Michaelis constant (Km) for cholesterol determined under two different experimental conditions were 4.5 and 6.7 X 10(-4) M. The enzymatic activity was remarkably inhibited by various metal salts such as FeC1(3), FeSO4, AgNO3, etc. On the other hand, neither EDTA nor Fe-chelating agents had any inhibitory effect on the enzymatic activity, while other metal-binding agents, KCN and NaN3, caused significant inhibition. The enzyme activity was inhibited almost completely by N-bromosuccinimide and iodine but not p-chloromercuribenzoate. The highly purified enzyme did not require any external electron acceptors other than oxygen. In addition, the activity was not influenced by the addition of external electron donors. The enzyme showed a high substrate specificity for 3beta-hydroxysteroids and the relative oxidation rates were 100 for cholesterol, 91 for 5alpha-cholestan-3beta-ol, 83 for pregn-5-en3beta-ol-one, 80 for androst-5-en-3beta-ol-17-one, 64 for 5alpha-androstan-3beta-ol-17-one, ect. The oxidation of cholesterol by the enzyme was remarkably inhibited by the addition of 5alpha-cholestan-3beta-ol, 5alpha-cholestan-3-one, 5beta-cholestan-3beta-ol, 5alpha-cholestane3beta, 5alpha-doil or 5alpha-lanosta-8, 24-den 3beta-ol. These findings indicate the present enzyme belongs to the class of 3beta-hydroxysteroid oxidase but some of its physical and enzymatic properties obviously differ from those of 3beta-hyroxysteroid oxidase of Brevibacterium.

A method to screen anticholesterol substances produced by microbes and a new cholesterol oxidase produced by Streptomyces violascens.

A new method to screen anticholesterol substances produced by microbes was devised and the method was applied for cultured broth of Streptomyces. Only one strain, resembled to be Streptomyces violascens, out of more than 200 Streptomyces was determined to produce an anticholesterol substance by this screening method. Furthermore, a new method to measure the anticholesterol activity was established for fermentation and purification of the active component. The partially purified active component was proved to be a new enzyme which oxidized cholesterol to cholest-4-en-3-one. Nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate was not necessary for this enzymatic oxidation. It was found that the enzyme was active against the 3β-hydroxyl group on the steroid skeleton but not against the 3α-hydroxyl group by investigating the substrate specificity of the enzyme.

Protease inhibitors from Streptomyces violascens. 3. Purification and some properties of the inhibitors.

Protease inhibitors from Streptomyces violascens. 3. Purification and some properties of the inhibitors.

Protease inhibitors from Streptomyces violascens. II. Production of the inhibitors.

Production of protease inhibitors from Streptomyces violascens U 10 600 was studied in quiescent flasks, shake flasks, and fermentors. A starting pH in the medium between pH 6 and 7 was the most favourable for the process. Both trypsin and chymotrypsin inhibitors were produced. Maximum inhibitor activity was attained after 3 days of cultivation in a protein powder medium at 18°C with pH control to pH 5 during the latter part of the process.

Streptomyces wistariopsis sp. nov. A new violet-spored species

In a taxonomic study of 35 “violetspored” streptomycetes certain essential differences were encountered among the different members. When the organisms were classified according to their sporophore morphology and spore surface configuration, they formed two distinct groupings. One of the groups was already represented by the species Streptomyces violascens Gauze; however, the other group contained organisms heretofore undescribed in the taxonomic literature. A new species Streptomyces wistariopsis is, therefore, proposed and described to represent this group.