Background: The majority of studies on soil Aspergillus concern the isolation and characterization of the antimicrobial compounds produced by this organism. Our previous studies indicated an isolated Aspergillus strain soil to be of interest, and this subject is further investigated here. Method: Soil samples of various locations in Iran were collected. Extract from Aspergillus sp. culture was obtained using ethyl acetate fractionation. Antimicrobial activity testing was performed using broth microdilution assay against Escherichia coli, Candida albicans, and Staphylococcus aureus microorganisms. One metabolite PA3-d10 was isolated from these active extracts and identified using thin layer chromatography, preparative thin-layer chromatography, HPLC, 1HNMR (proton nuclear magnetic resonance), 2D NMR, and LC-MS (liquid chromatography-mass spectrometry). Results: According to morphological and biochemical properties as well as ITS rDNA sequencing, we identified an isolate of Aspergillus flavus. The ethyl acetate fraction of the fermentation medium containing membrane active metabolites showed antimicrobial effects against different bacterial and yeast indicator strains. One metabolite from these active extracts was finally identified. Conclusion: Membrane active fraction produced by Aspergillus strain in this research demonstrated antimicrobial activities against bacteria and yeast strains. Therefore, this metabolite can be considered as a potential antimicrobial membrane active agent.

Induction of hairy roots in Perovskia abrotanoides Karel, a medicinally important member of the Lamiaceae family, was evaluated using different strains of Agrobacterium rhizogenes and various carbon sources. An efficient Agrobacterium mediated transformation system was established for the first time for this plant species using the bacterial strains of ATCC15834, TR105, and R1000. Meanwhile, the effects of acetosyringone and carbon sources on both rooting and root biomass production were investigated and the potential of hairy root cultures for producing tanshinones was explored. Hairy root induction in the nodes of intact seedlings was found to be successful with all the bacterial strains tested but R1000 was found to be more effective than TR105. The highest transformation frequency belonged to the nodes infected with ATCC15834 (47.33%). Moreover, transformation frequency increased by up to 60.99% when 100 mM of acetosyringone was added. The strain ATCC15834 recorded the highest root dry and fresh weights per 50 mL of the nutrient medium (0.149 and 1.996 g, respectively) and root length (34.45 cm) in a medium containing 3% sucrose. However, R1000 induced maximum lateral root density in hairy roots (7.1 branches cm-1) in a medium containing 3% glucose. PCR analysis using rolC specific primer with an amplicon of 612 bp confirmed the transgenic nature of the hairy roots. Finally, HPLC analysis revealed that the cryptotanshinone and tanshinone IIA levels in the hairy roots induced by TR105 (clone T7) were 53.17±0.26 and 14.48±0.30 mg g-1 DW, respectively.

Sericostoma pauciflorumStocks ex Wight (Family Boraginaceae) used against cancer, diabetes and known to be health promoter. Callus cultures have been established from the stem explants on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L concentration of different growth hormone viz. kinetin (Kn), indole 3-acetic acid (IAA) and indole 3-butyric acid (IBA). At 6 weeks of age, these calli were harvested, dried and extracted successively in pet, ether, methanol and water. Extracts were dried, weighed (%) and analyzed for their bioefficacies. Antimicrobial activities were determined using agar well diffusion and antioxidant potentials by DPPH and FRAP methods. Among all the test extracts, the extract of stem callus raised on IBA found to be more effective whereas it’s pet. ether extract showed appreciable activity against both the test bacteria and fungi (S. aureus- IZ 14.00±0.57 mm and T. rubrum- 16.33±0.32 mm), followed by methanol extract (S. aureus- IZ 13.00±0.57 mm, A. niger and P. chrysogenum - IZ 16.66 mm in both). In antioxidant potentials, all aqueous extracts were more active where IBA and Kn extracts demonstrated 0.06 mg/mL IC50 value (% inhibition 93.30 and 92.70 respectively at 0.8 mg/mL concentration) with 366±6.69 and 343±3.34 ascorbic acid equivalent antioxidant potentials at 1 mg/mL concentration. Furthermore, the chemical profile of test extracts was carried out. The bioactive secondary metabolites, b-sitosterol and caffeic acid was isolated from culture tissue of 6 weeks-old callus, and their identification and confirmation was carried out by color reaction, TLC behavior and IR spectrum techniques.

Background & Objective: The emergence of antibiotic-resistant bacteria and limited treatment options for these infections is a challenging problem in the world. There is increasing interest in isolating bioactive molecules from actinobacteria and elucidating their antimicrobial mechanisms at the moment. The current study aimed to investigate the antimicrobial activities of a Nocardia strain (N4) that was previously isolated from the soil samples in Tehran, Iran. Materials & Methods: The cultural, morphological, and physiological characteristics of the strain were determined and the identity of the strain was confirmed using 16SrRNA gene sequencing. The strain was screened for antimicrobial metabolite production. The in vitro anti-microbial activity of the extracted metabolite was assessed by agar well diffusion and minimum inhibitory concentrations methods. Finally, the structure of the extracted metabolite was confirmed by HPLC and mass spectrometry methods. Results: The isolate shared 99% sequence similarity with Nocardia soli. The strain showed broad-spectrum antimicrobial activities against some pathogenic bacteria (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 700603, Salmonella Typhimurium ATCC 14028 and Shigella sonnei RI366) and fungi (Aspergillus niger ATCC 1015 and Aspergillus fumigatus ATCC 1022) in the agar well diffusion method. The ethyl acetate extract of the strain exhibited the highest antimicrobial activity against E. coli (MIC= 4. 67±, 3. 055 μ, g/mL) and K. pneumoniae (MIC=3. 33±, 1. 155 μ, g/mL). The metabolite showed high similarity with C2 1H28 N7 O14. Conclusions: The current study confirmed the presence of a bioactive metabolite in strain N4, suggesting that this may be a promising candidate for discovering bioactive components with broad antibacterial activities against a variety of pathogens.

Objective(s): This study explored Indonesian Actinobacteria which were isolated from Curcuma zedoaria endophytic microbes and mangrove ecosystem for new antimycobacterial compounds. Materials and Methods: Antimycobacterial activity test was carried out against Mycobacterium tuberculosis H37Rv. Chemical profiling of secondary metabolite using Gas Chromatography-Mass Spectroscopy (GC-MS) and High Resolution-Mass Spectroscopy (HR-MS) was done to the ethyl acetate extract of active strain InaCC A758. Molecular taxonomy analysis based on 16S rRNA gene and biosynthetic gene clusters analysis of polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) from InaCC A758 have been carried out. Bioassay guided isolation of ethyl acetate extract was done, then structural elucidation of active compound was performed using UV-Vis, FT-IR, and NMR spectroscopy methods. Results: The chemical profiling using HR-MS revealed that InaCC A758 has the potential to produce new antimycobacterial compounds. The 16S rRNA gene sequencing showed that InaCC A758 has the closest homology to Streptomyces parvus strain NBRC 14599 (99. 64%). In addition, InaCC A758 has NRPS gene and related to S. parvulus (92% of similarity), and also PKS gene related to PKS-type borrelidin of S. rochei and S. parvulus (74% of similarity). Two compounds with potential antimycobacterial were predicted as 1) Compound 1, similar to dimethenamid (C12H18ClNO2S; MW 275. 0723), with MIC value of 100 μ g/ml, and 2) Compound 2, actinomycin D (C62H86N12O16; MW 1254. 6285), with MIC value of 0. 78 μ g/ml. Conclusion: Actinomycin D has been reported to have antimycobacterial activity, however the compound has been predicted to resemble dimethenamid had not been reported to have similar activity.

Echium amoenum Fisch. & Mey. (Boraginaceae) is a very popular medicinal plant that is used as a tonic, tranquillizer, diaphoretic, and a remedy for cough, sore throat and pneumonia in traditional medicine of Iran. Callus culture of medicinal plants is one of the ways for production of secondary metabolites. In this study, we investigated callus culture of E. amoenum and the major secondary metabolite. The callus culture of E. amoenum was initiated and established from seed in MS media with three different ratio of plant growth regulatories: kinetin, 2, 4-D and NAA. Methanolic extracts of freeze-dried calluses was compared by TLC and HPLC. The major secondary metabolite was separated by preparative HPLC and this pure compound was elucidated by UV, IR, one and two dimensional 1H and 13C NMR and Mass spectroscopy. Rosmarinic acid was identified by various spectroscopic methods from callus culture of E. amoenum. Rosmarinic acid is widespread in the plant cell tissue culture of the Lamiaceae and Boraginaceae families, through insignificant quantities. Rosmarinic acid has an antimicrobial, antiviral, and anti-inflammatory effect, which makes it a valuable product for the pharmaceutical and cosmetic industries.