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Designing novel isolation methods could enhance the diversification of the available bacterial strains to biotechnology. In this study, the new ex situ and in situ cultivation methods are introduced for the isolation of actinobacteria. In the ex situ experiments, the soil suspension was spread on the isolation media located above some ordinary papers in immediate contact with the slurry of soil substrate and incubated for 16 weeks. The paper was wholly immersed in the cave soil for in situ cultivations, and the containers were buried under layers of soil in Hampoeil cave for 10 weeks. Fruit wrapping kraft coated paper, with 68.8% recovery of isolates, was a better choice in isolation of actinobacteria than other studied filter paper. Based on the molecular identification results, 19% of the isolates obtained from the in situ cultivation method had less than 98.5% similarity to known taxa of actinobacteria and potentially may represent new species. In contrast, in the standard cultivation method, 1.3% of the isolates had less than 98.5% similarity 16Sr RNA gene. This data shows that the introduced cultivation method is a promising technique for isolating less culturable or new actinobacteria., (© 2021. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

High values of agitation and temperature lead to stressful conditions in the fermentations of Lactococcus lactis due to its aero-tolerant and mesophilic nature. Here, the adaptive laboratory evolution (ALE) technique was applied to increase biomass and nisin production yields by enhancing L. lactis subsp. lactis robustness at higher growth temperature and aeration rates. In two separate ALE experiments, after 162 serial transfers, optimum agitation and growth temperature of L. lactis were shifted from 40 rpm and 30 °C to 200 rpm and 37 °C, respectively. Oxidative and acid resistance were enhanced in the evolved strain. Whole-genome sequencing revealed the emergence of five single-nucleotide polymorphisms in the genome of the evolved strain in jag, DnaB, ArgR, cation transporter genes, and one putative protein. The evolved strain of L. lactis in this study has more industrial desirable features and improved nisin production capability and can act more efficiently in nisin production in stressful conditions., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Biosurfactants are structurally a diverse group of surface-active molecules widely used for various purposes in industry. In this study, among 120 fungal isolates, M-06 was selected as a superior biosurfactant producer, based on different standard methods, and was identified as Mucor circinelloides on the basis of its nucleotide sequence of the internal transcribed spacer (ITS) gene. M. circinelloides reduced the surface tension to 26 mN/m and its EI 24 index was determined to be 66.6%. The produced biosurfactant exhibited a high degree of stability at a high temperature (121°C), salinity (40 g/L), and acidic pH (2-8). The fermentation broth's ability to recover oil from contaminated sand was 2 and 1.8 times higher than those of water and Tween 80, respectively. The ability of biosurfactant to emulsify crude oil in the sea and fresh water was 64.9 and 48% respectively. This strain could remove 87.6% of crude oil in the Minimal Salt Medium (MSM) crude oil as the sole carbon source. The results from a primary chemical characterization of crude biosurfactant suggest that it is of a glycolipid nature. The strain and its biosurfactant could be used as a potent candidate in bioremediation of oil-contaminated water, soil, and for oil recovery processes., Competing Interests: The authors have declared no conflict of interest., (© 2018 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background and Objectives: Annual incidence of infection with S. Typhi is estimated to be about 17 million cases worldwide. A systematic search among actinomycete isolates from soil of Iran aimed at finding active actinomycetes against the causative agent of typhoid fever, Salmonella Typhi was carried out during this study., Materials and Methods: Our anti-Salmonella screening program resulted in nine highly active actinomycete isolates. All nine antibiotic producing strains showed broad-spectrum antibacterial activity, as five strains showed antifungal activity as well. Based on microscopic morphology and cell wall analysis, all nine active actinomycete strains were representatives of the genus Streptomyces. Three of the producing strains including the isolates HG87, HG116 and HG443 with inhibition zone of >20 mm, were selected for further identification and investigation of cytotoxic effects., Results and Conclusion: None of the producing strains showed cytotoxicity on HEK and USSC cell lines, while strain HG116 showed excellent antitumor activity on T47D cancer cell lines. Isolates HG87, HG116 and HG443 can be distinguished from the related species by some phenotypic and biochemical characteristics. Our results demonstrate the broad-range biological activity exhibited by bioactive compounds of soil actinomycetes from Iran.

Actinomycetes are interesting as a main producer of secondary metabolites and industrial antibiotics from marine environments. A total of 44 strains of actinomycetes were isolated from Caspian Sea sediments at a depth of 5-10 m. Preliminary screening was done using cross-streak method against 2 gram-positive and 4 gram-negative pathogen bacteria. The most potent strains MN2, MN3, MN38, MN39, MN40, MN41, and MN44 were used to extract the antibacterial substances. The antibacterial activities were performed using Kirby-Bauer disk diffusion method. Potent actinomycetes were screened for hydrolytic exoenzymatic activities (amylase and protease). All of the 24 isolates were active against at least to one of the test organisms. The MN38 strain showed activity against Staphylococcus aureus (20.0±0.5mm), Bacillus subtilis (27.0±0.2 mm), and Escherichia coli (20.0±0.3 mm). The MN39 strain was also active against E. coli (23.0±0.4mm), B. subtilis (23.0±0.2mm), Klebsiella pneumonia (24±0.1mm), whereas, the MN3 strain showed activity against Pseudomonas aeruginosa (20.0±0.2mm). The results of this investigation revealed that the marine actinomycetes of Caspian Sea sediments were potent source of novel antibiotics and bioactive compounds.

Negative control of clavulanic acid by phosphate in Streptomyces clavuligerus DSM 738 suggests that a pho regulon may exist in this bacterium. S. clavuligerus PhoP was expressed with a C-terminal His-tag in Escherichia coli and purified. Binding of PhoP-His 6 to promoter fragments of phoRP/phoU and pstS was demonstrated in gel retardation experiments. These fragments contained direct 11 bp repeats resembling PHO boxes. The tentative consensus sequence, GKTCRHBBNSV, was used to search other potential PhoP target genes in the genomic sequence of S. clavuligerus. In total, the putative PHO binding sequence was found in promoter regions of 31 S. clavuligerus genes. Binding of PhoP- His 6 to the PHO box present in the promoter region of the phosphate transporter gene SSCG_07547 of S. clavuligerus was demonstrated. Furthermore, it was shown by real time PCR that decreased concentrations of phosphate do affect increased expressions of genes to which PhoP binds. These findings confirm that a pho regulon exists in S. clavuligerus.

Actinomycetes isolated from Iran soil habitats were tested for the capacity to produce compounds which can protect neurons from cell death generated by oxidative stress in NT2 neurons. Confirmation of our initial hit was accomplished via the determination of amyloid beta level using the enzyme-linked immunosorbent assay test. The most interesting amyloid beta formation inhibitor discovered in our study was a secondary metabolite which was produced by strain HM45. This bioactive strain was identified as a strain of Streptomyces antibioticus DSM 40234 using polyphasic approach. The strain HM45 was deposited in Deutsche Sammlung von Mikroorganismen und Zellkulturen as S. antibioticus DSM 41955 and University of Tehran Microorganisms Sollection as S. antibioticus UTMC 00105. This work is the first report on efficiency of an actinomycete metabolite in prohibition of neurons death caused by amyloid beta formation.

Clavulanic acid is produced industrially by fermentation of Streptomyces clavuligerus and researches have increased its production by strain improvement, recombinant DNA technology, and media composition and growth condition optimization. The main objective of this study was to increase the level of clavulanic acid production from Streptomyces clavuligerus (DSM 738), using UV irradiation. After incubation, the spores and aerial mycelia were scraped off the agar plate by a sterile loop. After passing through a cotton wool, the serially diluted spore suspension was spread on GYM- agar containing caffeine. The plates were irradiated with UV light, wrapped in aluminum foil and incubated. The colonies were sub-cultured again to express the mutations. An aliquot of the spore suspension prepared from the resulted culture was poured in GYM agar plates and incubated. The plates were overlaid with nutrient-agar containing penicillin G and Klebsiela pneumoniae, and incubated. The inhibition zone diameter was measured and compared with the wild type colony. Repeating this procedure, the overproducer mutants were selected. Concentration of clavulanic acid was determined by HPLC analysis. It was concluded that secondary metabolites, mainly antibiotics containing clavulanic acid, were produced about 6-7 days after the growth, and concentration of clavulanic acid was increased up to two-folds after UV mutagenesis.