Starch Casein Research Articles

Identification of Endophytic Actinomycetes Isolated From Hopea ferrea and Its Antibacterial Activity Against Cariogenic Bacterium

Dental caries affects around 36% of the world's population and results in the loss of primary teeth in about 530 million youngsters. It is described as the loss of the enamel layer of the tooth by acids generated by the activity of cariogenic bacterium such as Streptococcus mutans on carbohydrates. This study aims to identify bioactive endophytic actinomycetes as an antibacterial agent against S. mutans DSM 20523 and to identify the bioactive endophytic actinomycetes using the 16S rRNA sequencing approach. To determine the percentage zone of inhibition, three endophytes at 10-day-old culture have been streaked onto ISP2. Then, S. mutans was streaked perpendicular to the endophytes. Crude extracts from three endophytes have been prepared by using four different types of media namely Tryptic Soy broth, Kings B, International Streptomyces Project Medium No.1 (ISP1) and Starch Casein broth. These crude extracts proceeded to undergo disk diffusion and MIC assays against S. mutans. All three endophytes from streaked plates with the highest percentage zone of inhibition were identified using the 16S rRNA gene molecular approach for characterisation. Endophytic actinomycetes which were isolated from Hopea ferrea showed potent antibacterial activity against S. mutans. All endophytic actinomycetes isolates from streak plates showed good inhibitory activity against the S. mutans. The highest percentage of inhibition was shown by the PT9-13S2 isolate followed by PT9-8S2 and PT9-13W2. Furthermore, crude extracts of PT9-8S2 and PT9-13W2 (the highest ZOI) from TSB media showed inhibitory activities against S. mutans, from disk diffusion as well as minimum inhibitory assays. From the 16S rRNA molecular approach, endophytic actinomycetes PT9-8S2 was closely related to Streptomyces collinus while PT9-13S2 and PT9-13W2 were closely related to Streptomyces malachitospinus. From this study, endophytic actinomycetes showed a promising source as antibacterial agents against cariogenic bacterium S. mutans which is the main causative agent of dental caries.

Screening, Production and Characterization of Potential Lignocellulolytic Actinomycetes from Agricultural Field

Actinomycetes are a suitable microbial group for the synthesis of lignocellulose-degrading enzymes. Enzymes that may degrade organic material, including cellulose, hemicellulose, and lignin, are released by actinomycetes. The aim of this research was to isolate actinomycetes from Rajkot, Gujarat, India’s soil and evaluate the activity of their cellulase and Xylanase enzymes. Starch Casein Agar (SCA) was used to identify a total of 30 isolates of actinomycetes. A qualitative plate assay (CMC-Na, Congo red) revealed that the highest zone of catalysis for MMD1 was 36 mm. Five strains were discovered to be effective for quantitative quantification of endoglucanase utilising filter paper and CMC as substrates: MMD1, MMD2, MMD3, MMD4, and MMD8. Following MMD 1 (endoglucanase 5.4 IU; FPase 4.4 IU), MMD 2 (endoglucanase 4.5 IU; FPase 3.4 IU) has demonstrated considerable endoglucanase and FPase activity. Beechwood xylan was used to treat sugarcane bagasse in order to test Xylanase, and 45% of the xylan (hemicellulose) fraction was obtained. MMD1 and MMD2 measured the xylanase enzyme activity (4.8IU and 4.2IU) in quantitative and qualitative assays (34 mm and 22 mm for BWX and 32 mm and 14 mm for agro-waste xylan). The strain MMD1 was identified as Streptomyces chartreusis through morphological, biochemical, and finally molecular characterization by 16S rRNA sequencing. It was then submitted to NCBI GenBank with the accession number MT254830.

Comparing the population density, colony diversity, and phosphate solubilization ability between rice root endophytic and rice rhizospheric soil bacteria from organic wetland

Abstract Phosphate-solubilizing bacteria (PSB) play important roles in the agricultural systems. The present study aims to compare the population density, colony diversity, and phosphate solubilization ability between rice root endophytic bacteria (RRoEB) and rice rhizospheric soil bacteria (RRzSB) from organic wetland. By spread plate method, bacteria were isolated directly using Pikovskaya’s agar and indirectly through other media, namely Nutrient Agar (NA), Starch Casein Agar (SCA), Jensen, and Yeast Extract Mannitol Agar (YEMA). Each distinct colony based on the morphology on the respective media was screened for phosphate solubilization ability by subculturing on Pikovskaya’s agar. The phosphate solubilizing index (PSI) was measured from the halo zone formation. The results showed that by direct isolation, 6 RRoEB and 5 RRzSB were able to phosphate solubilization. By indirect isolation from NA, SCA, Jensen, and YEMA, 2, 3, 1, and 3 isolates of RRoEB and 4, 3, 1, and 3 isolates of RRzSB showed the capability as phosphate solubilizer. Although the highest colony diversity of PSB was obtained from direct isolation, however the highest PSI was yielded from indirect isolation, namely isolates J1R1 (3.53) and Y1S1 (2.48). The highest PSI from direct isolation was obtained by P1R5 (1.90) and P1S1 (1.51).

Cellulolytic potentials of aspergillus oryzae and streptomyces griseus isolated from waste dump soil in Nile University Of Nigeria, Abuja

The potential of using microorganisms as biological sources of industrially economic enzymes has stimulated interest in the exploitation of extracellular enzymatic activity in several microorganisms. The aim of this research is to assess the cellulose degrading potentials of two microorganisms, Aspergillus oryzae and Streptomyces griseus using cellulose Congo red agar media. Soil sample collected from waste dump was serially diluted and inoculated in starch casein agar and SDA to isolate S. griseus and A. oryzae respectively. To assess their potentials to utilize cellulose, each of the two microorganisms was inoculated on cellulose Congo-red media and incubated at 30 ºC for 7days. A zone of clearing around the colonies after incubation confirms the secretion of extracellular cellulase, and was used as an indication for cellulose utilization. The zone of clearing was measured with a meter rule. In the results obtained, both microorganisms demonstrated cellulose utilization ability with Aspergillus oryzae showing a zone of clearing of 30.50 ± 0.50 mm while Streptomyces griseus showed a wider zone of clearing of 60.00 ± 1.00 mm. The results indicate that both microorganisms can be potent producers of the enzyme cellulase, with Streptomyces griseus having a higher cellulase-producing ability.

Production, optimization and characterization of partially purified anti-mycotic compound from marine soil derived streptomycetes originating at unexplored region of Bay of Bengal, India

Sixty-eight morphologically distinct isolates of marine actinomycetes were derived from seashore, mangrove, and saltpan ecosystems located between the Palk Strait and Gulf of Mannar region, Bay of Bengal, Tamilnadu. Twenty-five (36.8%) isolates exhibited anti-mycotic activity against Candida albicans and Cryptococcus neoformans in preliminary screening, and 4 isolates with prominent activity were identified and designated at the genus level as Streptomyces sp. VPTS3-I, Streptomyces sp. VPTS3-2, Streptomyces sp. VPTSA1-4 and Streptomyces sp. VPTSA1-8. All the potential antagonistic isolates were further characterized with phenotypic and genotypic properties including 16S rRNA gene sequencing and identified species level as Streptomyces afghaniensis VPTS3-1, S. matensis VPTS3-2, S. tuirus VPTSA1-4 and S. griseus VPTSA1-8. In addition, the active fractions from the potential antagonistic streptomycetes were extracted with organic solvents by shake flask culture method and the anti-mycotic efficacies were evaluated. The optimization parameters for the production of the anti-mycotic compound were found to be pH between 7 and 8, the temperature at 30ᵒC, the salinity of 2%, incubation of 9 days, and starch and KNO3 as the suitable carbon and nitrogen sources respectively in starch casein medium.

Predominance and bioactive potential studies of endophytic Bacillus Sps. from marine algae of Palkbay costal region, India

The genus Bacillus is a widely distributed and prominent species that has been extensively studied for its numerous benefits in various industries, medical applications, and agriculture. In this study, endophytic Bacillus species were isolated from seven different algal species using various growth media such as Nutrient agar, starch casein agar, ISP 2, and ISP 4. Throughout the study period, a total of 36 bacterial strains with similar morphological characteristics were identified. Among the isolates, five bacterial strains were found to be predominant when grown on ISP 2 media. These five strains were further subjected to 16s RNA Sequencing to determine their taxonomic classification. The results revealed that all of these strains belonged to the genus Bacillus. However, the other endophytic strains could not be identified due to limited sequence homology with existing entries in the NCBI database. To gain further insights into the isolated Bacillus cultures, analytical studies were conducted. Firstly, the functional groups present in the cultures were identified using Fourier-transform infrared spectroscopy (FTIR). Additionally, the compounds present in the crude extracts of the cultures were analyzed using gas chromatography-mass spectrometry (GC-MS). The findings of this current research demonstrate the presence of Bacillus species among green and brown algae in the Palk Bay coastal range. Moreover, these Bacillus strains exhibit potential for various biotechnological applications. The study aims to establish the predominance of Bacillus species in the algae of the Palk Bay region in South India, thereby contributing to our understanding of the microbial ecology in this specific geographical area

Green extraction of puromycin-based antibiotics from Streptomyces albofaciens (MS38) for sustainable biopharmaceutical applications.

Background: Microbial secondary metabolites have shown promise as a source of novel antimicrobial agents. In this study, we aimed to isolate, characterize, and evaluate the antimicrobial activity of compound from a novel Streptomyces albofaciens strain MS38. The objective was to identify a potential bioactive compound with broad-spectrum antimicrobial properties. Methods: The isolated strain MS38 on starch casein agar was characterized using morphological, physiological, and molecular identification techniques. The compound was obtained from the fermented broth through extraction with n-butanol and further purification using silica gel column chromatography and high-performance liquid chromatography (HPLC). Structural elucidation was conducted using Ultraviolet (UV), Infrared (IR), nuclear magnetic resonance (NMR), and mass spectrometry (MS) techniques. The antimicrobial activity was evaluated using the agar well diffusion method and the microplate Alamar blue assay (MABA). Results: The isolated strain MS38 was identified as novel S. albofaciens based on morphological characteristics and confirmed by 16S sequences analysis and MALDI-TOF MS. The compound obtained from the fermented broth exhibited substantial antimicrobial activity against a variety of pathogenic bacteria and fungi. Structural analysis revealed a complex chemical structure with characteristic functional groups indicative of potential antimicrobial properties. The compound demonstrated strong activity against both Gram-positive (Staphylococcus Spp.) and Gram-negative (Klebsiella pneumoniae and Escherichia coli) bacteria, as well as fungi, including Candida albicans and Trichophyton rubrum. Conclusion: This study successfully isolated and characterized a bioactive compound from a novel S. albofaciens MS38. The compound exhibited significant antimicrobial activity against a range of pathogenic microorganisms. These findings underscore the importance of exploring microbial biodiversity for the discovery of novel antimicrobial agents. This study contributes to the growing knowledge of microbial secondary metabolites with potential therapeutic value.

Biochemical characterization of actinomycete from Namibia rocky crest mountainous soil and analyzing their bioactive metabolites for antagonistic effect against human respiratory pathogens.

the present study aimed at isolating and characterizing actinomycete from unexplored Windhoek rocky crest mountainous soil and extracting bioactive metabolites as possible therapeutics against common life-threatening Streptococcus pneumonia (S. pneumonia) and Stachybotrys chartarum (S. chartarum). chemotaxonomy and biochemical methods were used to identify the isolates. The solvent extraction method was used to extract bioactive compounds. Agar overlay and disc diffusion methods were used to determine the antimicrobial activity of isolates and extracted bioactive metabolites against S. pneumonia and S. chartarum. The antioxidant activity of the extracted bioactive metabolites was determined using 2.2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method with ascorbic acid as a positive control. Comparison between groups was done using a Two-way ANOVA, followed by Bonferroni post-test. three distinct isolates from 3 soil samples were identified on starch casein agar and distinguished using biochemical tests. All three isolates showed strong inhibitory activity against S. pneumonia with average growth inhibition zones between 18.0±1.00 and 27±0.00 mm p< 0.005. All isolates showed potent inhibitory activity against S. chartarum with the average inhibition zones ranging between 42.0±1.00 and 48±0.00 mm, p< 0.005. The chloroform extracts showed potent DPPH activity of up to 73± 1.41%. growth conditions and extraction solvents can influence the antimicrobial and antioxidant properties of bioactive metabolites.

Screening and Molecular Characterization of Antibacterial Secondary Metabolite Producing Actinomycetes from Soils of Eastern Mountain Regions of Nepal

Antibacterial secondary metabolite is a bioactive compound like antibiotic that can be considered a substance, produced by one microorganism, which in low concentration inhibits the growth of other microorganisms. Actinomycetes, slow-growing gram-positive bacteria, are the major sources of bioactive compounds. This study aimed to screen and identify antibacterial secondary metabolite-producing actinomycetes by sequencing the 16S rRNA method (molecular identification) from the soils of the mountain region of eastern Nepal. Starch casein agar (SCA) medium was used for the isolation of actinomycetes which were confirmed by primary screening and secondary screening. Identification of presumptive genera was done based on macroscopic, microscopic, and biochemical characteristics and confirmed by sequencing their 16S rRNA genes. The antibacterial compound was produced by culturing the potential isolate in starch casein broth. Using organic solvents such as ethyl acetate, n-butanol, chloroform, dichloromethane, and methanol, the chemical was recovered from the fermented broth. TLC performed the antibacterial substance characterization. Only 9 (13.6%) of the 66 actinomycetes isolates showed antibacterial activity against test microorganisms. Only one of the nine isolates, M3, had antibacterial activity in primary screening against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli, Salmonella Typhi, Shigella spp., and Pseudomonas aeruginosa) test bacteria. M3 was chosen for secondary screening due to its strong antibacterial activity. The minimum inhibitory concentration (MIC) of crude antibacterial substances was found to be 2.5 mg/mL against test organisms. According to the TLC chromatogram, the isolate produced only one compound with an Rf value of 0.81, completely distinct from the spot formed by gentamicin (standard), which had an Rf value of 0.89. The isolates were considered Streptomyces spp., a distinct taxonomic group based on characterization by macroscopic, microscopic, biochemical, physiological, and molecular techniques. This study concluded that Mountain regions are the reservoir of antibiotic- producing actinomycetes. Streptomyces is the most common genus.

Neurotoxic Acrylamide Mitigation By L-Asparaginase Enzyme Extracted From Actinomycetes Species.

Acrylamide is a chemical that is generated in starchy foods, coffee, and bread, etc during high-temperature cooking processes, such as frying, roasting, and baking and is a major food safety concern. Acrylamide toxicity targets the nervous and reproductive systems, causing symptoms like muscle weakness and numbness therefore there is a need to solve this problem by biological method. Actinomycetes are an excellent source of L-asparaginase, an enzyme which specifically catalyses the breakdown of amino acid L-asparagine to aspartic acid and ammonia. If starchy food products are not treated with L- asparginase enzyme then at high temperature L-asparagine present in food converts to acrylamide. Hence, water and soil samples were obtained from Uran, Sai, and Panvel regions and Actinomycetes were isolated onto Starch Casein Agar plates. Biochemical analyses revealed negative results for TSI, nitrate reduction, and citrate but positive for catalase and starch hydrolysis. Noteworthy enzymatic activity at 2.66µg/ml/min was observed. Optimization was performed with various parameters. Production and purification of L-asparaginase by submerged fermentation, followed by ammonium precipitation, dialysis, and subsequent application to pretreat potatoes. After incubation and frying, high-performance liquid chromatography analysis exhibited a reduction in acrylamide levels. The study's meticulous approach, from isolation to application, addresses critical food safety issues, marking a significant contribution.

Biodiversity and biological applications of marine actinomycetes—Abu-Qir Bay, Mediterranean Sea, Egypt

BackgroundThe ability of actinomycetes to produce bioactive secondary metabolites makes them one of the most important prokaryotes. Marine actinomycetes are one of the most important secondary metabolites producers used for pharmaceuticals and other different industries. ResultsIn this study, the promising actinomycetes were isolated from Abu-Qir Bay. Four different media named as starch nitrate, starch casein, glycerol asparagine, and glycerol glycine were used as a preliminary experimental media to study the role of the medium components on the counts of actinomycetes in sediment samples. The results indicated that starch casein medium reported the highest counts (30–63 CFU/g) in all the tested sites. Lower counts were detected on starch nitrate and glycerol asparagine. On the other hand, glycerol glycine medium gave the lowest counts (15–48 CFU/g). Abu-Qir8 harbored the highest average count of actinomycetes (63 CFU/g), followed by Abu-Qir1 (48 CFU/g). The lower counts were detected in Abu-Qir5 and Abu-Qir7 (26 and 29 CFU/g, respectively). A total of 12 pure obtained actinomycetes isolates were subjected to morphological, physiological, and biochemical characterization. The selected actinobacterial isolates were subjected to numerical analysis, and the majority of isolates were grouped into four main clusters (A, B, C, & D), and each of them harbored two isolates; additionally, four isolates did not cluster at this similarity level. Isolate W4 was carefully chosen as the most promising pigment and antimicrobial agent’s producer; the produced pigment was extracted and optimized by statistical experiments (PBD & BBD) and was tested for its anti-inflammatory activity. The results showed anti-inflammatory effect and prevented the denaturation of BSA protein at a concentration much higher than the safe dose and increased with increasing the pigment concentration. ConclusionMarine actinomycetes play a vital role in the production of novel and important economic metabolites that have many industrial and pharmaceuticals applications. Streptomyces genera are the most important actinomycetes that produce important metabolites as previously reported.

Screening of bacteria isolates from earthworm cast for antibacterial activities

The healthy operation of the soil ecosystem is significantly influenced by earthworms. Various medical treatments have included the use of earthworms. This study isolated and screened bacterial species from earthworm cast for antibacterial activities. Two (2) bacteria species were isolated from the earthworm cast by culturing on starch casein agar using pour plate techniques. The isolates were identified as Streptomyces and Actinomadura species while the test isolates include Klebsiella specie, Salmonella shigella and Staphylococcus aureus strains. This study revealed that Streptococcus and Actinomadura metabolite exhibited variable degrees of antimicrobial activities against the test isolates. The highest in-vitro antimicrobial activity is (12.0mm) was exhibited by the Streptococcus metabolite at the highest concentration of 100mg/ml against Staphylococcus aureus, while the least antimicrobial activity (8.0mm) was exhibited at the concentration of 80mg/ml and 40mg/ml against Salmonella species and Staphylococcus aureus. Actinomadura metabolite was effective against Staphylococcus aureus at the highest concentration of 100mg/ml. However there was significant difference observed in antibacterial activity of the metabolites when compared to the standard antibiotic (Gentamicin) (p&lt;0.05). From the minimum inhibitory concentration (MIC) test result, Streptococcus metabolite demonstrated greater activity on Salmonella specie at the range of 3.12mg/ml and a lower activity on Staphylococcus aureus and Klebsiella specie at the range of 12.5mg/ml each. The Actinomadura metabolite demonstrated greater activity on Salmonella species at the range of 1.56mg/ml. Based on the findings of this research, earthworm (Pheretima posthuma) is a good source of antibacterial agents that can be identified and extracted as source of cheap medicines to control serious infections.

The Influence of Soil’s Physicochemical Properties and Land Use Systems on the Abundance of Actinomycetes Populations

Aims: In this study, we assessed the effect of physicochemical properties of soils from different land use systems on the population of Actinomycetes.&#x0D; Study Design: A cross sectional survey design in line transect sampling was used in collection of samples from land use systems. A 4×6×3 factorial experiments laid in completely randomized design was used in determining Actinomycetes population in land use system and media type culture.&#x0D; Place and Duration of Study: The study was conducted in Meru south sub-county, Kenya, between January 2019 and July 2019.&#x0D; Methodology: Cultural growth of Actinomycetes was determined using four different media type culture. The physicochemical properties of soil were evaluated using standard laboratory methods. The data obtained were analyzed using analysis of variance and significantly means were separated using Least Significance Difference at α = 0.05 in SAS version 9.4. Correlation analysis was used to determine the relationship between soil physicochemical properties and Actinomycetes population. Actinomycetes population varied significantly (p &lt; 0.05) among the different land use systems.&#x0D; Results: The study's findings revealed significant variations (p &lt; 0.05) in Actinomycetes populations across the different land use systems. The type of media used significantly influenced the growth and proliferation of Actinomycetes colonies, with the Starch casein medium exhibiting the highest number of colonies (3.4 × 105 cfu/ml of soil sample) and the modified Luria Bertani (M1) medium showing the lowest (1.7 × 105 cfu/ml). The Actinomycetes populations demonstrated significant variations (p &lt; 0.05) among the different land use systems. Correlation analysis revealed a moderate positive correlation (r = 0.63407; p &lt; 0.0001) between Actinomycetes population and soil pH, as well as weaker positive correlations (r = 0.3375; p = 0.012) between soil moisture content and Actinomycetes population, and available potassium and Actinomycetes load (r = 0.31483; p &lt; 0.0001).&#x0D; Conclusion: The study concludes that the population of Actinomycetes was affected by soil physicochemical properties and consequently land uses.

Structural elucidation of a compound extracted from Streptomyces Sp.NLKB45.

A Streptomyces Sp. NLKB45 was isolated from the Nellore coastal region. After checking antimicrobial and antifungal activity the strain was subjected to fermentation in starch casein broth medium. HPLC was used to purify the culture filtrate after it had been extracted with ethyl acetate. The extract showed antibacterial and anticancer properties. The extract was processed and the compound isolated was further analyzed using FTIR, mass spectroscopic method and one- and two-dimensional NMR techniques. Based on the data gathered the compound was identified as 7-((5-methylpyridin-2-ylamino) (phenyl) methyl) quinolin-8-ol with a weight-average molecular weight of about 341 Da.

GC-MS Based Metabolomic Profiling of Streptomyces clavuligerus Isolated from Ocimum gratissimum Rhizosphere

Streptomyces clavuligerus is a member of the Actinobacteria family primarily known for its production of clavulanic acid antibiotic. The need for identification of new antimicrobials led to the identification of volatile components of S. clavuligerus metabolites using GC-MS. The isolate was obtained from Ocimum gratissimum rhizosphere using starch casein agar, and identified using molecular typing. The preliminary antibacterial screening of the isolate was carried out using some indicator bacteria from wound sites and urinary tract infection. Its bioactive metabolites were obtained using sub-merged fermentation over a four-day period, and the volatile compounds identified using GC-MS. The organism showed significant (p&lt;0.05) inhibition on Pseudomonas aeruginosa and Escherichia coli. Metabolomics study revealed the presence of compounds of alkanone and alkene functional groups. Eicosene was the major antimicrobial compound identified. Likewise, a non-antimicrobial, steroidal metabolite – pregnelonone, was also dominant in the metabolite mixture produced by the organism. Identifying volatile constituents of microbial metabolites may be a route for obtaining new antimicrobials and the GC-MS is a useful tool for achieving such aim.

Ceropegia rupicola, Acokanthera schimperi, Jatropha variegate Antimicrobial, Cytotoxicity and Phytochemical

In Asian countries, a variety of medicinal plants are utilized in folk medicine for hemostatic and antiseptic. The methanolic extracts of Jatropha variegata, Acokanthera schimperi, and Ceropegia rupicolawere tested for the following purposes: first, isolation of endophytic fungi, second, antimicrobial and cytotoxicity activity, and third, phytoconstituents of the methanolic extract using the GC-Mass technique. The plants came from a variety of Yemeni locations. Endophytic fungus strains were isolated and identified using starch casein and potato dextrose agar medium. Methanol was used to extract the stem and leaves of the plants. The agar well diffusion method was used to test the antibacterial properties of the produced methanolic extracts against four Gram-negative bacteria, three Gram-positive bacteria, and one species of yeast. The IC50 values for the most promising extracts are calculated. The methanolic extracts were also subjected to a phytochemical screening by GC-Mass. A total of forty fungal endophyte isolates were isolated. Fusarium spp. (15), Phomaspp. (6), Alternaria alternata (5), White Sterile mycelia (6), Dark Sterile mycelia (6), Cochlioboluslunatus (1), and Mucor spp. (1). were found throughout the identification process. The highest yield extract was obtained from Jatropha variegate (45.4%), followed by Acokanthera schimperi (32.3%), and finally Ceropegia rupicola (20.5%). Only Gram-negative bacteria were found to have antibacterial activity, and the most effective extract was Acokanthera schimperi against the majority of microorganisms tested (8–18mm). The methanolic extracts of Jatropha variegate had the best antibacterial activity against Klebsiella pneumoniae ATCC 13883015, with a zone of inhibition (45 mm). Furthermore, Ceropegia rupicola extract had the highest inhibitory zone against Candida albicans ATCC 700012 (20mm). In addition, the IC50 results showed that the Jatropha variegate extract on A549 had an IC50 of 84.1ug/ml and a PC3 of 39.4ug/ml. The IC50 for Ceropegia rupicola extract was 49.4ug/ml in A549 cells and 34ug/ml in PC3 cells. Also, the screening for phytochemicals of the three plant extracts revealed the presence of many chemicals such as ethanol, formic acid, and others that may be responsible for the observed actions.

Production of Fibrinolytic Enzyme by Soil Actinobacteria

Thrombotic diseases are increasingly becoming among the prominent causes of death across the globe. Researchers are now turning attention towards fibrinolytic enzymes as potential alternative treatment for thrombolytic diseases. This present study focused on the production of extracellular fibrinolytic enzyme from soil Actinobacteria and evaluation of its hydrolytic activity on blood clot. The Actinobacteria was isolated from compost soil of semi-arid climate in Nigeria. Bacterial isolation was achieved using starch casein agar. Potent fibrinolytic enzyme producing Actinobacteria were identified and subjected to enzyme production using submerged fermentation method. The interactive effects of incubation time, temperature, pH and media components on enzyme production were analysed. Extracellular fibrinolytic enzyme produced by the selected Actinobacteria was partially purified by ammonium sulphate precipitation and subsequently assayed for blood clot lysis activity. Results of these studies indicated that fibrinolytic enzyme was produced optimally at pH 8 and temperature of 40 oC after 72 hour of fermentation. Partially purified fibrinolytic enzyme was able to degrade blood clot comparable to the positive control. These results shows that soil Actinobacteria of unexplored semi-arid climate of Nigeria present a prospect in search of novel microorganisms with potentials in the production of fibrinolytic enzyme that can serve as an alternative blood clot buster in treating thrombolytic diseases.

Antiproliferative activity of antimicrobial peptides and bioactive compounds from the mangrove Glutamicibacter mysorens.

The Glutamicibacter group of microbes is known for antibiotic and enzyme production. Antibiotics and enzymes produced by them are important in the control, protection, and treatment of chronic human diseases. In this study, the Glutamicibacter mysorens (G. mysorens) strain MW647910.1 was isolated from mangrove soil in the Mangalore region of India. After optimization of growth conditions for G. mysorens on starch casein agar media, the micromorphology of G. mysorens was found to be spirally coiled spore chain, each spore visualized as an elongated cylindrical hairy appearance with curved edges visualized through Field Emission Scanning Electron Microscopy (FESEM) analysis. The culture phenotype with filamentous mycelia, brown pigmentation, and ash-colored spore production was observed. The intracellular extract of G. mysorens characterized through GCMS analysis detected bioactive compounds reported for pharmacological applications. The majority of bioactive compounds identified in intracellular extract when compared to the NIST library revealed molecular weight ranging below 1kgmole-1. The Sephadex G-10 could result in 10.66 fold purification and eluted peak protein fraction showed significant anticancer activity on the prostate cancer cell line. Liquid Chromatography-Mass Spectrometry (LC-MS) analysis revealed Kinetin-9-ribose and Embinin with a molecular weight below 1 kDa. This study showed small molecular weight bioactive compounds produced from microbial origin possess dual roles, acting as antimicrobial peptides (AMPs) and anticancer peptides (ACPs). Hence, the bioactive compounds produced from microbial origin are a promising source of future therapeutics.

Aktivitas Daya Hambat Isolat Actinomycetes dengan Lama Fermentasi yang Berbeda terhadap Pertumbuhan Klebsiella pneumoniae

Actinomycetes are filamentous, Gram-positive bacteria widely distributed in soil and can produce secondary metabolites useful as antibiotics. The synthesis of secondary metabolites of Actinomycetes is influenced by the growth conditions of each isolate, including the fermentation period. Klebsiella pneumoniae is an opportunistic bacterial pathogen causing lower respiratory tract infection. Objectives: To determined the inhibition activity of Actinomycetes with different fermentation periods against the growth of K.pneumoniae. Methods: Actinomycetes were isolated from soil in Kebun Raya Bogor and grown on Starch Casein Agar (SCA) medium before being fermented in liquid media consisting of 2% of mannitol, 2% of peptone, and 1% of glucose with an incubation time of 6,7, and 8 days. An antibacterial activity test was conducted on Mueller Hinton Agar (MHA) medium using the well diffusion method. Results: The activity of Actinomycetes isolates with fermentation duration of 6,7 and 8 were able to inhibit the growth of K. pneumoniae with an average inhibition zone of 4,46 mm; 4,94 mm; and 5,04 mm, respectively. Actinomycetes' inhibitory mechanism ranges from inhibition of cell wall synthesis to nucleic acid and protein synthesis. One Way ANOVA test showed significant differences between the inhibition zone produced by each fermentation group. Conclusion: Actinomycetes isolates have antibacterial activity against the growth of K. pneumoniae with the highest activity achieved by Actinomycetes with a fermentation time of 8 days.Keywords: Actinomycetes, antibacterial, fermentation, Klebsiella pneumoniae

Presence of Actinomycetes in agarwood tissues of Aquilaria crassna: A preliminary study

Agarwood is a valuable resin produced inside certain tree species of the family Thymalaeaceae distributed in the Asian region. Agarwood production occurs as a defense mechanism when the trees are under physical or biological stresses. However, the formation of agarwood resins in significant extractable quantities due to natural stress conditions is rare; therefore, the farmers use various methods to induce its formation artificially. Certain fungal species such as Fusarium and Aspergillus become more popular among them to produce high-quality agarwood. However, studies are rare on using other microbial organisms such as Actinomycetes, which exhibit properties of both bacteria and fungi. Among the agarwood-producing species, Aquilaria crassna is one of Asia's most commonly planted species for agarwood production. This species was introduced to Sri Lanka in 2012 for mid and lower elevations of the wet zone. Due to the lack of studies on agarwood resin formation by non-fungal microbial methods, the present study attempted to identify the presence of Actinomycetes species in agarwood resinous tissues of A. crassna. Agarwood resinous tissue samples were collected from four plantations in the wet zone of the country. Surface sterilized, small sized tissues were placed on starch casein agar medium and incubated at room temperature for ten days. Using the morphological and microscopic characteristics, it was possible to identify Nocardia, Psuedonocardia, and three Streptomyces species with varying abundance. The species level should be confirmed using molecular analysis, and their potential for agarwood resin formation inducement should be tested by re-inoculating to the healthy A. crassna trees.