Telomere and Telomerase in Cancer

The linear ends of chromosome are protected by specialized ribonucleoprotein (RNP) termed as telomere. These specialized terminal elements with tandem repeated sequence are the protective cap that alleviate end replication problem and cell senescence. The telomere length maintenance is essential to avoid cell death and apoptosis. Telomere shortening has been related to chronic stress due to several factors, which include not only psychological stress but also diseases such as cardiovascular diseases and cancer. Telomerase enzyme which maintains telomere length is the major factor responsible for evading cell death. Telomere length maintenance and telomerase expression put together are the prerequisite for immortality, an essential character for cancer cells. Understanding the mechanism of telomere and telomerase functions paves way for eradicating the diseases such as cancer

Mycoremediation of Benzo[a]pyrene by Pleurotus ostreatus in the presence of heavy metals and mediators

Benzo[a]pyrene is considered as a priority pollutant because of its carcinogenic, teratogenic and mutagenic effects. The highly recalcitrant nature of Benzo[a]pyrene poses a major problem for its degradation. White-rot fungi such as Pleurotus ostreatus can degrade Benzo[a]pyrene by enzymes like laccase and manganese peroxidase. The present investigation was carried out to determine the extent of Benzo[a]pyrene degradation by the PO-3, a native isolate of P. ostreatus, in the presence of heavy metals and ligninolytic enzyme mediators. Modified mineral salt medium was supplemented with 5 mM concentration of different heavy metal salts and ethylenediaminetetraacetic acid. Vanillin and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (1 and 5 mM) were used to study the effect of mediators. Results indicated that P. ostreatus PO-3 degraded 71.2 % of Benzo[a]pyrene in the presence of copper ions. Moderate degradation was observed in the presence of zinc and manganese. Both biomass formation and degradation were severely affected in the presence of all other heavy metal salts used in the study. Copper at 15 mM concentration supported the best degradation (74.2 %), beyond which the degradation progressively reduced. Among the mediators, 1 mM 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) supported 78.7 % degradation and 83.6 % degradation was observed under the influence of 5 mM vanillin. Thus, metal ion like copper is essential for better biodegradation of Benzo[a]pyrene. Compared to synthetic laccase mediator like 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate), natural mediator such as vanillin may play a significant role in the degradation of aromatic compounds by white-rot fungi

PURIFICATION AND CYTOTOXICITY STUDY OF LOVASTATIN FROM SOIL FUNGI

  Objective: The objective of the present study is to evaluate the anticancer potential of lovastatin obtained from fungal source.Methods: About 15 fungal cultures were isolated from soil samples collected from Bharathiar University, India, and all are identified and characterized through microscopic characterization. Lovastatin producing capability was confirmed through bioassay against Saccharomyces cerevisiae, and the ability of selected fungus to produce lovastatin was further confirmed by high-performance liquid chromatography. Maximum lovastatin producing fungi were further selected for purification (overloaded elution chromatography) and characterization done using inhibition rate (IR). 5-diphenyl tetrazolium bromide (MTT) assay using A549 cell line was performed for antitumor activity evaluation.Results: Among the 15 isolates, Aspergillus flavus exhibited the maximum zone of inhibition (1.5 cm) against the test organism through solid-state fermentation. The resemblance in retention time (RT) of peak shown in chromatograms of standard lovastatin (RT=25.1 minutes) and sample (RT=25.1 minutes) were similar. This confirmed the presence of lovastatin in the selected fungal isolate (A. flavus). The presence of two functional groups in lovastatin C=O and O-H was confirmed by IR spectrum 50% of cell lysis was observed in MTT assay.Conclusion: Lovastatin obtained from soil fungi is capable of producing lovastatin in good proportions. Obtained fungal lovastatin exhibited significant antitumor activity against A549 cell line. Like other biological derivatives, lovastatin from soil fungi had greater potential in anticancer activity, and further biosynthetic pathway insights in their production can improve the yield which aid in large scale production

SCREENING AND PRODUCTION OF ANTICARCINOGENIC ENZYME FROM ESCHERICHIA COLI CTLS20: L - ASPARAGINASE

Objective: The objective of this study was attempted to screen the production of L-asparaginase from bacteria isolated from soil samples and its enzymatic activity.Methods: Screening of L-asparaginase was performed using phenol red indicator growth medium from which the positive strains were chosen based on the colour change. The enzyme production of L-asparaginase was established by submerged fermentation followed by the molecular detection of the efficient bacterial strains.Results: The enzyme production was undertaken by submerged fermentation with the evaluation of enzymatic activity and protein content. This revealed that the strain Escherichia coli CTLS20 produced a higher yield of L-asparaginase (30.22 IU/mg), 16.91 µg/ml of protein with the specific activity of 1.787 IU/mg when compared with other bacterial strains. The efficient bacterial strains were also confirmed by 16S rRNA sequence as Escherichia coli, Acinetobacter baumnnii, Klebsiella pneumoniae and the phylogenetic tree construction revealed the evolutionary relationship of the bacterial strains.Conclusion: This study indicated that the bacterial strain E. coli CTLS20 had the ability for the higher production of L-asparaginase. This novel higher yielding bacterial asparaginase is highly desirable as better alternatives in cancer therapy.Keywords: Soil, L-asparaginase, Submerged fermentation, E. coli, Phylogenetic tre

PRODUCTION AND PURIFICATION OF ANGIOTENSIN-CONVERTING ENZYME INHIBITOR BY SELECTED BACTERIAL STRAIN FOR CANCER THERAPY

Objective: The present study was planned to explore safer, innovative and economic Angiotensin-converting enzyme inhibitors (ACEi) from beef extract by the action of a proteolytic Micrococcus luteus. Cytotoxicity of the stable peptide was predicted using MCF-7 cell line in vitro.Methods: ACEi was purified by sequential steps of ethanol precipitation, ion exchange column chromatography (MonoQ) and gel filtration column chromatography (Sephadex G25). The apparent molecular mass was determined by SDS-PAGE. The anticancer property was analyzed by studying the cytotoxicity effects of angiotensin converting enzyme inhibitor using Breast cancer MCF-7 cell linesResults: The peptide was purified and molecular mass was determined as 4.5 kDa. The IC50 value of peptide was found to be 59.5 µg/ml. The DNA fragmentation was not observed in the treated cells. The purified peptide has demonstrated to induce apoptosis of cancer cell. The results proved that the peptide has the ability to be used for cancer therapy.Conclusion: The presence of ACE inhibition activities in the fermentation of beef extract using Micrococcus luteus has been investigated. The Peptide has been determined as an active compound that inhibited the activity of ACE. These properties indicate the possibilities of the use of purified protein as a potent anticancer agent.Keywords: Angiotensin-converting enzyme inhibitors, Micrococcus luteus, Anti-proliferative, Anti-metastatic, MCF-7 cell line, Anticancer activity

ISOLATION OF ANGIOTENSIN-CONVERTING ENZYME INHIBITOR PRODUCING BACTERIA FROM COW MILK

Objective: To evaluate the potential of protease producing organism for the production of Angiotensin I–converting enzyme (ACE) inhibitor by fermentation of various protein substrates.Methods: Bacterial strains were isolated from cow milk collected in Coimbatore, Tamil Nadu, India by using serial dilution technique, plated on nutrient agar medium. The identity of the strain was ascertained by 16s rRNA gene sequencing method and was submitted to the NCBI GenBank nucleotide database. Various substrates were screened for ACE inhibitor production by the fermentation with the isolated strain.Results: The isolated coded as BUCTL09, which showed a significant zone of clearance was selected and identified as Micrococcus luteus (KF303592.1). Among the seven substrates, only beef extract fermented broth showed an inhibition of 79% and was reported as the best substrate.Conclusion: In the search for non-toxic, and economic ACE inhibitors as an alternative to the synthetic drugs, many natural ACE inhibitors have been isolated from a microbial source. In the present study, isolate BUCTL09 was selected for the production of ACE inhibitor from the beef extract. Findings from this study lead us to investigate this potent ACE inhibitor further for its biological properties and to explore the impending efficacy of the ACE inhibitor which may conceivably be developed into a prospective drug

Isolation and Growth Characterization of the Fresh Water Algae Chlorosarcinopsis Eremi on Different Growth Media

In the present study the freshwater algae Chlorosarcinopsis eremi was isolated from the water sample of Kovai Kutralum. The pure culture of the algae was obtained using serial dilution technique followed by streak plate method. The species was observed under light microscopy and identified with the help of algal manuals and also by sequencing 18S rRNA genes. The growth of the algae, C. eremi was characterized in different growth media. The different growth media’s include Bold’s Basal medium, Modified Bourrelly medium, BG 11 medium, Half strength Chu10 medium and Modified Hoagland’s medium. The growth was measured by observed optical density of the biomass and chlorophyll “a” content. The experiment was carried out three times and mean values and standard deviation were calculated. From the results it is concluded that growth of C. eremi was maximum in Bold’s basal medium when compared with other media tested

Production of Xylanase by Litter Degrading Fungal Species Using Agro-industrial Wastes as Substrates by Solid State Fermentation

ABSTRACT Xylan is a heterogeneous polysaccharide which are mainly constituted by β-1,4-linked-D-xylopyranose. Xylan forms a main constituent of hemicellulose, a plant polysaccharide. The most important enzyme in the xylan biodegradation is the endo-1, 4-β-xylanase (EC 3.2.1.8) that releases xylopyranose units. Agricultural wastes are found in staggering amounts in our country. These residues represent one of the most energy-rich resources available and when not properly discharged or used, add to environmental pollution. In the present study, thirty fungal species of various genera were isolated from litter soil. The objective of the study is to find the fungus that produces high titres of endo-β-1, 4-xylanase in solid state fermentation using various agro-industrial wastes as substrate. Three fungal species were selected and solid state fermentation was carried out. Aspergillus nidulans and Trichoderma viride produced higher enzyme production of 561.75 U/g and 446.25 U/g respectively when grown on rice bran. Penicillium frequentans produced the highest among the three producers when grown on paddy straw to an extent of 735 U/g. These results indicate that agro-industrial wastes can be used to produce xylanase and thus reduce the risk of environmental pollution

Alcalase Potato Protein Hydrolysate-PPH902 Enhances Myogenic Differentiation and Enhances Skeletal Muscle Protein Synthesis under High Glucose Condition in C2C12 Cells

Sarcopenia is an aging associated disorder involving skeletal muscle atrophy and a reduction in muscle strength, and there are no pharmaceutical interventions available thus far. Moreover, conditions such as hyperglycaemia are known to further intensify muscle degradation. Therefore, novel strategies to attenuate skeletal muscle loss are essential to enhance muscle function and thereby improve the quality of life in diabetic individuals. In this study, we have investigated the efficiency of a potato peptide hydrolysate PPH902 for its cytoprotective effects in skeletal muscle cells. PPH902 treatment in C2C12 cells showed the dose-dependent activation of the Akt/mTOR signalling pathway that is involved in skeletal myogenesis. According to Western blotting analysis, PPH902 induced the phosphorylation of Akt, mTOR proteins and induced the myogenic differentiation of C2C12 myoblasts in a differentiation medium. The phosphorylation myogenic transcription factor Foxo3A was also found to be increased in the cells treated with PPH902. In addition, treatment with PPH902 ameliorated the high glucose induced reduction in cell viability in a dose-dependent manner. Moreover, the number of myotubes in a differentiation medium reduced upon high glucose challenge, but treatment with PPH902 increased the number of differentiated myotubes. Further, the phosphorylations of AMPK and mitochondrial-related transcription factors such as PGC-1α were suppressed upon high glucose challenge but PPH902 treatment restored the protein levels. We demonstrate, for the first time, that a specific potato peptide has a therapeutic effect against sarcopenia. In addition, PPH902 improved the myogenic differentiation and their mitochondrial biogenesis and further improved myogenic protein and inhibited muscle protein degradation in C2C12 cells challenged under a high glucose condition