A study on Tornabea scutellifera (Lichenized Ascomycete, Lecanorales) in northeastern Iran

The present study shows the morphology, anatomy and chemistry of Tornabea scutellifera occurring in north eastern Iran . Most thalli are corticolouse, even though its thalus also shows some degree of vegetation on rocky substrates

Purification and characterization of an extracellular alkaline cold-adapted serine metalo-protease from the cold tolerant bacterium, Stenotrophomonas sp. BTR88

444-450Proteases are the most economical enzymes in biotechnology and industry. Nowadays, a lot of attention is being paid to extremophiles microorganisms owing to the diversity of their enzymes. One hundred and Six proteolytic bacteria were isolated from Binaloud Mountain; one of them (strain BTR88) was selected as the best producer of extracellular protease and was used for further studies. This bacterium belongs to Stenotrophomonas sp., which were identified by the 16S rDNA sequence. Maximal protease production was detected at the beginning of exponential growth phase in the presence of starch and skim milk at 20°C and pH 9. This protease was purified to electrophoretic homogeneity with a fold: 27.5, yield: 33% and specific enzyme: 12.6 U/mg. SDS-PAGE and zymography analyses revealed a protein band of 22 kDa. The maximum activity was at pH 9 and in the range of 20-30°C; while, the enzyme exhibited a broad range of activity from 20-80°C as well as the pH of 5-10. Enzyme inhibition in the presence of phenylmethane sulfonyl fluoride (PMSF) and Ethylenediaminetetraacetic acid (EDTA) showed that the purified enzyme belongs to serine metallo-enzymes. Kinetic parameters of km, Vmax and kcat for the cold tolerant enzyme were determined to be 7.2 mg/mL, 1.45 mM/min and 33.2 sec−1, respectively. The characteristics of activity in cold and alkaline conditions and the broad range of pH and temperature suggest that serine metalloprotease has potential use in the detergent industry

Purification and characterization of an extracellular alkaline cold-adapted serine metalo-protease from the cold tolerant bacterium, Stenotrophomonas sp. BTR88

Proteases are the most economical enzymes in biotechnology and industry. Nowadays, a lot of attention is being paid to extremophiles microorganisms owing to the diversity of their enzymes. One hundred and Six proteolytic bacteria were isolated from Binaloud Mountain; one of them (strain BTR88) was selected as the best producer of extracellular protease and was used for further studies. This bacterium belongs to Stenotrophomonas sp., which were identified by the 16S rDNA sequence. Maximal protease production was detected at the beginning of exponential growth phase in the presence of starch and skim milk at 20°C and pH 9. This protease was purified to electrophoretic homogeneity with a fold: 27.5, yield: 33% and specific enzyme: 12.6 U/mg. SDS-PAGE and zymography analyses revealed a protein band of 22 kDa. The maximum activity was at pH 9 and in the range of 20-30°C; while, the enzyme exhibited a broad range of activity from 20-80°C as well as the pH of 5-10. Enzyme inhibition in the presence of phenylmethane sulfonyl fluoride (PMSF) and Ethylenediaminetetraacetic acid (EDTA) showed that the purified enzyme belongs to serine metallo-enzymes. Kinetic parameters of km, Vmax and kcat for the cold tolerant enzyme were determined to be 7.2 mg/mL, 1.45 mM/min and 33.2 sec−1, respectively. The characteristics of activity in cold and alkaline conditions and the broad range of pH and temperature suggest that serine metalloprotease has potential use in the detergent industry

A Potent Antifungal Activity by the Marine Streptomyces albidoflavus sp. ADR10 from the Caspian Sea Sediment: Optimization and Primary Purification

Fungal infections are an evolving public health challenge due to their antimicrobial resistance and the growth of immunocompromised populations. Aquatic environments, the largest ecosystem on earth, are recently considered as a source for the production of bioactive compounds. Marine actinomycetes are considered for their potential to produce novel bioactive metabolites like antifungal compounds. In this study, strain ADR10 was obtained from the sediment sample of the Caspian Sea and its 16S rDNA gene sequence analysis suggested that the isolate belongs to Streptomyces albidoflavus. The preliminary cross-streak and double-layer agar screening revealed that the isolate has potent activity against pathogenic fungi, i.e. Aspergillus niger, Candida albicans, Fusarium oxysporum, and Penicillium crustosum. One-factor-at-a-time and Response surface methodology (RSM) was employed to evaluate the effects of six parameters (carbon source, initial pH, inoculation volume, NaCl concentration, nitrogen source, and temperature) on the production of antibiotics in the basal starch casein broth medium. The maximum antibiotic activity was achieved at the initial pH 7.05, sucrose 1.17 g l-1, malt 0.2 g l-1, temperature 30 ºC, inoculation size 5.0% v/v, and NaCl 1% w/v after 121.1 hours. Through the optimization experiments, antifungal activity was enhanced 2.7-fold.  Ethyl acetate showed the highest antibiotic extraction capacity from the fermentation media compared with dichloromethane, hexane, and chloroform. The preliminary purified antibiotic by thin layer chromatography (ethyl acetate/ mobile petroleum phase) showed a more significant growth inhibition zone than nystatin (100 μg mL-1) against Candida albicans. This study underlines the potential of the marine actinomycete for the identification of novel antifungal agents

In silico designing and expression of novel recombinant construct containing the variable part of CD44 extracellular domain for prediagnostic breast cancer

Abstract Background CD44, as a tumor‐associated marker, can be used to detect stem cells in breast cancer. While CD44 is expressed in normal epithelial cells, carcinoma cells overexpress CD44. Aims In the current study, we designed a recombinant protein that included the variable component of the CD44 (CD44v) extracellular domain to apply in clinical diagnosis of breast cancer. Methods A total of 100 CD44v amino‐acid residues were determined, and the structure was examined using bioinformatics tools. The construct was inserted into the PET28a vector and transformed in E. coli BL21(DE3). A nearly 12 kDa fusion protein was obtained by Ni‐NTA affinity metal chromatography. Recombinant CD44v was examined by Western blotting, ELISA, and immunohistochemistry (IHC) assays. Results The findings revealed that the structure of rCD44v was stable, and its antigenic domain was exposed. The recombinant CD44v was confirmed by western blotting, and the presence of antibodies against recombinant CD44v protein in the patient's serum was detected by the ELISA. Our data demonstrated a link between CD44v serum levels and the prevalence of breast cancer. Conclusion Assessments of antiCD44v antibodies with rCD44v could be a useful tool for identifying breast cancer in its early stages, which can lead to better outcomes

Cytotoxic and antioxidant capacity of camel milk peptides: Effects of isolated peptide on superoxide dismutase and catalase gene expression

Peptides from natural sources such as milk are shown to have a wide spectrum of biological activities. In this study, three peptides with antioxidant capacity were identified from camel milk protein hydrolysate. Pepsin and pancreatin were used for hydrolysis of milk proteins. Ultrafiltration and reverse-phase high-performance liquid chromatography were used for the concentration and purification of the hydrolysate, respectively. Sequences of the three peptides, which were determined by matrix-assisted laser desorption/ionization time-of-flight spectrophotometry, were LEEQQQTEDEQQDQL [molecular weight (MW): 1860.85 Da, LL-15], YLEELHRLNAGY (MW: 1477.63 Da, YY-11), and RGLHPVPQ (MW: 903.04 Da, RQ-8). The 3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to evaluate the cytotoxicity of these chemically synthesized peptides against HepG2 cells. In vitro analysis showed antioxidant properties and radical scavenging activities of these peptides on 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)+, O2–, and OH– free radicals. HepG2 cells were treated with YY-11 peptide for 48 hours, and the expression of superoxide dismutase and catalase genes was examined using real-time polymerase chain reaction. The results revealed a significant increase in the expression of superoxide dismutase and catalase genes in treated HepG2 cells

Anti-leishmanial activity of Brevinin 2R and its Lauric acid conjugate type against L. major: In vitro mechanism of actions and in vivo treatment potentials.

Leishmaniasis, as a major health problem in tropical and sub-tropical areas in the world, needs novel, safe, nontoxic and plausible therapeutic solutions for its control. As a part of innate immune system, natural antimicrobial peptides have a potential to be used as new generation of antibiotics especially after persistent resistance of conventional antimicrobial agents. Brevinin 2R, a member of Defensin families of host defense peptides, showed promising effects against bacterial and fungal infections as well as cancerous cell lines. In the current research, the anti-leishmanial effect of Brevinin 2R and its lauric acid conjugate was investigated against Leishmania major (L. major) parasite. The data revealed that, conjugation of fatty acid to Brevinin 2R, strengthen its effect on L. major promastigotes as well as toxicity and hemolytic effect. These peptides showed anitleishmanial activity through cell membrane disruption and changes in the electrical and mitochondrial membrane potential. No signs of apoptosis induction or caspase activation were detected. Despite its hemolytic and cytotoxic effect in in vitro conditions, lauric acid- Brevinin 2R (L- Brevinin 2R) did not show site specific adverse reactions in animal model. Treatment course with L- Brevinin 2R in the L. major infected mice exhibited decreased parasite load in the lymph nodes adjacent to the infected site despite cytokine production profile and footpad swelling data