Streptomyces Badius Research Articles

Bioformulation of Azotobacter spp. and Streptomyces badius on the productivity, economics and energetics of wheat (Triticum aestivum L.)

Wheat production is directly linked to energy inputs of high yielding varieties, chemical fertilizer and mechanization. In spite of their benefits, fertilizers are associated with high energy consumption. Thus there is need to develop energy-efficient agricultural systems with low input energy compared with the output of food. The only alternate to increase and/or sustain the productivity by developing a system in which nutrients are added through combined use of organic sources biofertilizers and PGPR's. Biofertilizers comprised of efficient strains of nitrogen fixing or phosphate solubilizing microorganism can play pivotal role in this regard. Thereupon the present investigation was planned with combination of chemical fertilizer and biofertilizer to find out the best combination and to get maximum profit with energy saving. The field experiment was carried out at two locations with nine different chemical and biofertilizer combination in randomized complete block design. Maximum grain yield was observed in 87.5% N and 87.5% P + Azotobacter spp and Streptomyces badius (seed treatment) at both the locations. Results indicated that Azotobacter spp and Streptomyces badius may be used synergistically as a potential source with reduction of chemical fertilizer to 12.5–25% without compromising yields.

The potency of actinomycetes extracts isolated from Pramuka Island, Jakarta, Indonesia as antimicrobial agents

Abstract. Setiawati S, Nuryastuti T, Sholikhak EN, Lisdiyanti P, Pratiwi SUT, Sulistiyanti TR, Ratnakomala S, Jumina, Mustofa. 2021. The potency of actinomycetes extracts isolated from Pramuka Island, Jakarta, Indonesia as antimicrobial agents. Biodiversitas 22: 1104-1111. Actinomycetes are one of the Gram-positive bacteria which are widely distributed and produce many secondary metabolites including those known as antibiotics, antifungals, anticancer, and antimalarials agents. The secondary metabolites of actinomycetes are abundant, which include many active compounds that have been identified because of the large diversity in the actinomycetes phylum. This study aimed to identify and screen collected by Indonesian Culture Collection (InaCC) from Bojong Gede and Pramuka Island, Jakarta, Indonesia as antibacterial and antifungal agents. Primary screening was done on 16 actinomycetes isolates by well-agar diffusion method. Antimicrobial activity was tested by using micro broth dilution methods to determine minimum inhibitory concentration (MIC). Molecular identification into level genera and species was determined by 16S rRNA gene sequencing. Out of 16 actinomycetes isolates used, 4 isolates have activity against Candida albicans ATCC 10231, Staphylococcus aureus ATCC 6538, Bacillus subtilis BTCC B-612, and Escherichia coli BTCC B-614, specifically InaCC A758, InaCC A759, InaCC A760 and InaCC A765 isolates. InaCC A758 have highest antimicrobial activity against mentioned microbial with MIC value at 50 μg/mL, 6.25 μg/mL, 31.25 μg/mL and 3.125 μg/mL, respectively. Three genera were found from the samples: i.e. Streptomyces (80%), Microbispora (13%) and Nocardia (6%). Based on 16S rRNA gene identification, the active isolates of actinomycetes InaCC A758, InaCC A759, InaCC A760 and InaCC A765 were similar to Streptomyces badius, Streptomyces olivaceus, Streptomyces sanyensis, and Nocardia otitidiscaviarum, respectively. The secondary metabolites of actinomycetes extracts from Pramuka Island can be potentially developed as antifungal and antibacterial agents.

Dinactin from a new producer, Streptomyces badius gz-8, and its antifungal activity against the rubber anthracnose fungus Colletotrichum gloeosporioides

Colletotrichum gloeosporioides is a main cause of rubber anthracnose, which results in very large losses for the natural rubber industry. In this study, an actinomycete strain gz-8 was isolated and had strong antagonistic activity against C. gloeosporioides, with an inhibition rate of 72.5 %. Strain gz-8 was identified as Streptomyces badius. Three active compounds were separated from S. badius gz-8 and identified as feigrisolide B, feigrisolide C and dinactin according to the mass spectrometry and NMR-spectra results. In the three compounds, dinactin exhibited the best antifungal activity against C. gloeosporioides, with an EC50 value of 2.55 μg/mL, and its minimum inhibitory concentration was 44 μg/mL. Dinactin had broad inhibitory activities against nine other pathogenic fungi, and it also had an obvious control effect on rubber anthracnose comparable to that of chlorothalonil. Dinactin could inhibit the conidiogenesis and spore germination of C. gloeosporioides. This report will contribute to understanding the antifungal activity of dinactin against C. gloeosporioides.

Efficiency of application of modern sanitation supplies in beekeeping

The article presents the results of studies of the effect of commercial disinfectants registered in Ukraine and allowed for use in beekeeping, as well as a comparative characteristic of their effectiveness in the disinfection of wooden beehive structures. The research was conducted on the basis of the departments of hygiene, sanitation and general veterinary prevention and technology of production and processing of products of small animals of the Lviv National University of Veterinary Medicine and Biotechnology named after S.Z. Gzhytskyi and SLW “Biolab” Food and Veterinary Diagnostic Laboratory (Ostroda, Poland). For research in the spring and summer period 4 groups of test objects were formed on the principle of analogues. Distilled water was used to control disinfection (test group 1). In the second experimental group for disinfection used “Brovadez-20” (active substance benzalkonium chloride 0.5%), in the third experimental group гру “Vetox-1000” (active substance hypochlorite 0.3%) and in the fourth experimental group – “Sumerian silver”(active ingredient of 10% citrate of Argentum citrate). In a comparative analysis of the effectiveness of the disinfectants of the II, III and IV groups, the growth of gram-positive (Streptomyces badius, Curtobakterium) and gram-negative bacteria (Xanthomonas hyacinthi, Pseudomonas alcaligenes, Acidovorax defluvii) was detected, respectively, among the endophores. In the current literature, information on the threat to the health of the honey bee detected by microorganisms is missing. According to the research, certain differences between the effectiveness of disinfectants have been established. It was found that the total microbial number (BMI) was 0.15% in group II, 0.16% in group III and up to 0.10% in group IV relative to control. It was found that the Sumer silver disinfectant at 10% concentration provides the best bactericidal effect (P < 0.001). In a comparative intergroup analysis of the effectiveness of disinfectants, it was found that in the conditions of the experiment, the level of bactericidal activity was the highest in the experiment with the SMC in the case of the use of “Sumer silver” was 32.3% and 34.4% better compared to “Brovades-20” and “Vetox-1000”, respectively. The use of “Sumerian silver” allows you to get the desired result 10 and 4 times faster than the disinfectants “Brovadez-20” and “Vetox-1000” respectively.

Actinobacterial community in Shuanghe Cave using culture-dependent and -independent approaches.

Karst caves, considering to be the "arks" of biodiversity, often contain high levels of endemism. In the present study, the actinobacterial community in Shuanghe Cave, the longest cave in Asia, was analyzed for the first-time using culture-dependent and -independent (16S rRNA amplicon sequencing) approaches. The amplicon sequencing analysis revealed a broad taxonomic diversity in Shuanghe Cave, including 19 phyla (predominantly Actinobacteria) and 264 different genera. While the culture-dependent method got the unrepresentative but supplemental result, a total of 239 actinomycetes were isolated and were identified to seven genera based on culture features and 16S rRNA tests. Among the three habitats (soil, rock soil, and bat guano), the dominant phyla did not differ significantly, while the dominant genus community varied among different habitats, and the richness in soil and rock soil samples was higher than that in bat guano. Furthermore, 16 isolate strains showed antimicrobial activity, especially, the strain S142 (Streptomyces badius) and S761 (Actinoplanes friuliensis) exhibited the most promising activity against various pathogens. Overall, this work showed the abundant bacterial diversity and the antimicrobial potential of the isolates from the Shuanghe Cave.

Mitigating the impact of climate change on wheat by use of liquid microbial inoculants under different planting dates

The present investigation was undertaken to study the response of agronomic traits of wheat to different planting dates and liquid inoculants of Azotobacter sp. and Streptomyces badius by mitigating the impact of climate change. The field experiment was conducted at Regional Research Station, Bathinda, Punjab during two consecutive years. The main plots treatment consisted of five planting dates (25th October, 10th November, 25th November, 10th December and 25th December) and four liquid inoculant treatments (T1: 100% RDF (control), T2: 100% RDF+seed bacterization with liquid inoculants, T3: 75% RDF+seed bacterization with liquid inoculants and T4: 75% RDF+seed bacterization with liquid inoculants+foliar application of liquid inoculants at critical stages of wheat) in sub-plots. The treatments of 20 combinations were tested in split plot design with three replications. Maximum plant height, effective spike length, grains per spike, 1000-grain weight and highest grain yield were attained by the crop planted on 10th November. Amongst the liquid inoculant treatments, T4 treatment was found superior in all growth and yield attributing parameters at each date of planting followed by T3, T2 and T1. The T4, T3 and T1 treatments resulted in 4.79, 3.47 and 1.61% more grain yield over T1. Thus, it can be concluded that liquid inoculants of Azotobacter sp. and S. badius can enhance growth and yield of wheat by mitigating the impact of climate change. This combination may be used as an efficient PGPR for wheat production.

Isolation, molecular identification and statistical optimization of culture condition for a new extracellular cholesterol oxidase-producing strain using response surface methodology

The current work details the screening of about 100 isolates from various soil samples, from which 1 isolate was finally selected based on the productivity of cholesterol oxidase. Further biochemical identification tests and 16S rRNA gene sequencing identified this isolate as Streptomyces badius. A preliminary culture media optimization was carried out using the initial screening method of Plackett-Burman. Then, a Box-Behnken design was employed to investigate the optimum concentrations of medium components and interactive effects of main variables on cholesterol oxidase production. The regression analysis showed a significant coefficient of determination (R 2) value (91 %), which was in close agreement ensuring a satisfactory adjustment of the proposed model. Maximal enzyme production (2.38 U/mL, i.e., approximately more than 100 % activity in the basal medium) was obtained at: temperature 35 °C; Tween 20 0.1 %; pH 6.5 and yeast extract 0.15 %. This two-stage statistical approach provided rapid identification and integration of key medium parameters for Streptomyces sp., resulting in high cholesterol oxidase production.

Effect of biodelignification of rice straw on humification and humus quality by Phanerochaete chrysosporium and Streptomyces badius

The effect of biodelignification of rice straw by two different ligninolytic organisms, Phanerochaete chrysosporium (white-rot fungus) and Streptomyces badius (actinomycetes), on humus quality was investigated during a 56-day incubation at 30 °C. Lignin degradation, the release of humic extract (HE), humic acid (HA) and fulvic acid (FA), E4/E6 ratio of HA, and humification index (HI, HA/FA) were measured during the incubation. Lignin was degraded by both organisms, but to different extents. Lignin was degraded to 41% and 31% by P. chrysosporium and S. badius, respectively. HE released by P. chrysosporium and S. badius were, respectively, 2.10 and 2.13 times larger than that in the control at the maximum values. A significant correlation between lignin degradation and humus-related parameters involving HA fraction showed that both organisms are converting lignin to humic substances.

Effect of different Streptomyces strains on pine wood as seen by Py–GC/MS : Implications for mechanical pulping

Pyrolysis/gas chromatography/mass spectrometry was used to elucidate the ability of streptomycetes to modify pine wood under solid-state fermentation. The pyrograms of sound and decayed wood showed the typical pyrolysis products derived from carbohydrates and lignin. In general, the relative abundance of the former was lower in treated woods than in sound wood. Several major peaks released from guaiacyl units of lignin were also revealed. Although some differences could be observed in the relative abundance of the released guaiacyl units from lignin between sound and treated woods, a clear decrease in the lignin-derived products by the action of the microorganisms was not observed. Nevertheless, an increase in the relative abundance of the most of guaiacyl units with a higher oxidation degree was mainly detected in woods treated by the strains Streptomyces UAH 7, Streptomyces anulatus, Streptomyces UAH 48, Streptomyces avermitilis and Streptomyces badius. These results suggest an oxidative action of these strains on the lignin of pine wood, which was corroborated by a decrease in the (aldehyde/ketone + acid) ratio. In addition, the breakdown of the alkyl chain linkages by the strain Streptomyces UAH 7 could be also observed. Analytical pyrolysis allow to demonstrate for first time the oxidative attack of selected Streptomyces strains on lignin moiety of pine wood, which is an important requirement for biomechanical pulping purposes.

Effect of pre-sowing seed inoculation with liquid biofertilizers on fodder yield and quality of sorghum (Sorghum bicolor)

An experiment was carried out during the rainy (kharif) seasons of 2016 and 2017 at 2 locations of Punjab (Ludhiana and Bathinda), to study the effect of N-fixer (Azotobacter species) and P-solubilizers (Sphingobacterium species, Burkholderia species and Streptomyces badius) inoculations and their interactions, in the presence of recommended dose of fertilizer (RDF) on SL 44 single-cut forage sorghum [Sorghum bicolour (L.) Moench]. A to- tal of 9 liquid microbial inoculant combinations were used as seed treatment before sowing. All the microbial treat- ments along with chemical fertilization resulted in non-significant numeric increase in yield and growth of sorghum at both the locations in 2 consecutive years. The dual inoculation, i.e. Azotobacter species + Burkholderia species along with RDF resulted in 21.5% and 16.8% percent increase in crude protein (CP) and in-vitro dry-matter digest- ibility (IVDMD) over the control, observed from 2 locations pooled mean respectively. Similar combination of biofertilizer with recommended dose of fertilizer resulted in lower acid detergent fibre (ADF) and neutral detergent fibre (NDF) content of 40.40% and 53.53% respectively. The hydrocyanic acid (HCN) production was also noticed to be lowered down with the use of liquid dual inoculum of Azotobacter species + Burkholderia species. Green fod- der yield (GFY) had positive significant correlation with all yield, growth and quality parameters, whereas negative correlation with HCN (0.09), ADF (0.34) and NDF (0.18). Dry-fodder yield (DFY) exhibited the highest signifi- cant correlation coefficient with IVDMD (0.92) followed by CP (0.71). The results of correlation also indicated that the plant height should be given due importance as it showed significant correlation with dry-fodder yield. There- fore, there was synergetic effect of nitrogen-fixing and phosphate-solubilizing microbial inoculants on fodder qual- ity improvement of sorghum.

Decolorisation of the polymeric dye Poly R by Streptomyces viridosporus T7A

AbstractThe ability of a range of actinomycetes to decolorise the polymeric dye Poly R was investigated. Three well characterised lignocellulose‐degrading actinomycetes, Streptomyces viridosporus, Streptomyces badius and Thermomonospora mesophila were found to be the most efficient at decolorising Poly R, with a maximum decolorisation rate of 0.10 unit per day. Extracellular fractions taken from S. viridosporus grown on a variety lignocellulose‐related substrates were also capable of decolorising Poly R, indicating that dye decolorisation was not merely due to biomass uptake. The activity of extracellular fractions from straw‐grown cultures of S. viridosporus was three to six times greater than those from other substrates examined. Purification of this dye‐decolorising activity from S. viridosporus using anion exchange chromatography revealed the presence of a single active fraction. This was confirmed using gel permeation chromatography which estimated the molecular mass of the decolorising protein to be approximately 30000 Da.

Towards elucidation of the lignin degradation pathway in actinomycetes

Six biodegradative actinomycete strains were grown on a dimeric model lignin compound of the β-aryl ether type. Although only two strains, Thermomonospora mesophila and Streptomyces badius, utilized the compound as a carbon and energy source and produced substantial amounts of monomeric products, all of the strains could demethylate the substrate and oxidize Cα on the phenylpropane side-chain. Streptomyces sp. EC1 produced small amounts of aromatic acids and unidentified lignin-derived products when grown on straw. This organism also produced cell-bound demethylase requiring H2O2 and Mn2+, protocatechuate 3,4-dioxygenase and β-carboxymuconate decarboxylase activity in response to growth on low-molecular-mass aromatic compounds but not lignocellulose or its polysaccharide components. Extracellular peroxidase and catalase activity were detected in all of the strains. These data are used to propose a scheme by which actinomycete attack of the lignin component of plant biomass can be envisaged.

Production of an extracellular polyethylene-degrading enzyme(s) by Streptomyces species.

Extracellular culture concentrates were prepared from Streptomyces viridosporus T7A, Streptomyces badius 252, and Streptomyces setonii 75Vi2 shake flask cultures. Ten-day-heat-treated (70 degrees C) starch-polyethylene degradable plastic films were incubated with shaking with active or inactive enzyme for 3 weeks (37 degrees C). Active enzyme illustrated changes in the films' Fourier transform infrared spectra, mechanical properties, and polyethylene molecular weight distributions.

Immunologic relatedness of extracellular ligninases from the actinomycetesstreptomyces viridosporus t7a andstreptomyces badius 252

Four isoforms of the extracellular lignin peroxidase of the ligninolytic actinomycete Streptomyces viridosporus T7A (ALip-P1, P2, P3, and P4) were individually purified by ultrafiltration and ammonium sulfate precipitation, followed by electro-elution using polyacrylamide gel electrophoresis. Three of the purified peroxidases were compared for their immunologic relatedness by Western blot analysis using a polyclonal antibody preparation produced in rabbits against pure isoform P3. The anti-P3 antibody was also tested for its reactivity towards a lignin peroxidase from the white-rot fungus Phanerochaete chrysosporium and another ligninolytic actinomycete Streptomyces badius 252. Results showed that peroxidases ALip-P1 through ALip-P3 are immunologically related to one another. The peroxidases of S. badius, but not the peroxidase of P. chrysosporium, also reacted with the antibody, thus indicating that the lignin peroxidases of S. viridosporus and S. badius are immunologically related. Based upon its specific affinity, lignin peroxidase isoform ALip-P3 of S. viridosporus was readily purified using an anti-P3 antibody affinity column.

Biotransformation of coal by ligninolytic Streptomyces

The chemistry of coal liquefaction by Streptomyces viridosporus, Streptomyces setonii and Streptomyces badius was studied. These ligninolytic actinomycetes solubilized lignite and bituminous coals when coal particles were placed on the surfaces of mycelial lawns on agar culture media. On agar, liquefaction proceeded with the formation of clear to black water-soluble products. Up to 40% of some coals was solubilized in 3 weeks. Lignite coals were also solubilized in liquid shake flask cultures. Coal solubilization by S. viridosporus was oxidative, as shown by Fourier transform infrared spectroscopy and elemental analysis. Compared to the starting coals, the soluble products were enriched in oxygen, nitrogen, sulfur, and ash, but were lower in hydrogen relative to carbon. Insoluble coal residues were less altered than the liquefied products, but they were higher in ash content than the starting coals. Chemical analysis of culture filtrates showed that some low molecular weight products were released, but most of the soluble products were acid precipitable and probably polymeric. Lignite coal solubilization by the S. viridosporus did not require induction and in liquid media solubilization was always accompanied by a significant increase in medium pH. When medium pH was not allowed to become basic, extracellular coal solubilizing activity was negligible. Extracellular solubilizing activity was heat stable, and ultrafiltration experiments showed that the solubilizing agent was of less than 10 000 molecular weight. Coal solubilizing activity of S. viridosporus culture supernatants could be eliminated by lowering their pH, but the activity could be recovered by raising the pH back up to above neutral. Phosphate and Tris buffers of varying strength and pH showed that nonbiological solubilization of lignite coal occurred under basic conditions. The combined data show that biological lignite coal solubilization by these Streptomyces is mediated by alkaline products that the cells release into the growth medium. These findings confirm and expand an earlier report by others that coal solubilization by Streptomyces setonii may be nonenzymatic.

β-Lactamase genes of Streptomyces badius, Streptomyces cacaoi and Streptomyces fradiae: Cloning and expression in Streptomyces lividans

Genes encoding extracellular β-lactamases (EC 3.5.2.6) of Gram-positive Streptomyces badius, Streptomyces cacaoi and Streptomyces fradiae have been cloned into Streptomyces lividans. The β-lactamase gene of S. badius was initially isolated on a 7 kb BamHI fragment and further located on a 1300 bp DNA segment. An 11 kb BamHI fragment was isolated encompassing the S. cacaoi β-lactamase gene, which was subcloned to a 1250 bp DNA fragment. The β-lactamase gene of S. fradiae was cloned on an 8 kb BamHI fragment and mapped to a 4 kb DNA segment. Each of the three BamHI fragments encompassing the β-lactamase genes hybridized to a BamHI fragment of the corresponding size in chromosomal DNA from the respective strain used for cloning. The activities of the three β-lactamases were predominantly found to be extracellular in the S. lividans recombinants. The S. badius and S. cacaoi β-lactamases exhibited a 10–100-times lower activity in S. lividans, whereas the S. fradiae β-lactamase showed an approximately 10-fold higher activity in the cloned state, compared with the activities found in the original strains.

The importance of fertilizers in raising soil fertility

Wheat production is directly linked to energy inputs of high yielding varieties, chemical fertilizer and mechanization. In spite of their benefits, fertilizers are associated with high energy consumption. Thus there is need to develop energy-efficient agricultural systems with low input energy compared with the output of food. The only alternate to increase and/or sustain the productivity by developing a system in which nutrients are added through combined use of organic sources biofertilizers and PGPR's. Biofertilizers comprised of efficient strains of nitrogen fixing or phosphate solubilizing microorganism can play pivotal role in this regard. Thereupon the present investigation was planned with combination of chemical fertilizer and biofertilizer to find out the best combination and to get maximum profit with energy saving. The field experiment was carried out at two locations with nine different chemical and biofertilizer combination in randomized complete block design. Maximum grain yield was observed in 87.5% N and 87.5% P + Azotobacter spp and Streptomyces badius (seed treatment) at both the locations. Results indicated that Azotobacter spp and Streptomyces badius may be used synergistically as a potential source with reduction of chemical fertilizer to 12.5–25% without compromising yields.