Enhanced production of prodigiosin by Serratia marcescens MO-1 using ram horn peptone

This work addresses the production of prodigiosin from ram horn peptone (RHP) using MO-1, a local isolate in submerged culture. First, a novel gram-negative and rod-shaped bacterial strain, MO-1, was isolated from the body of the grasshopper (Poecilemon tauricola Ramme 1951), which was collected from pesticide-contaminated fields. Sequence analysis of 16S rDNA classified the microbe as Serratia marcescens. The substrate utilization potential (BIOLOG) and fatty acid methyl ester profile (FAME) of S. marcescens were also determined. The effect of RHP on the production of prodigiosin by S. marcescens MO-1 was investigated, and the results showed that RHP supplementation promoted the growth of MO-1 and increased the production of prodigiosin. A concentration of 0.4% (w/v) RHP resulted in the greatest yield of prodigiosin (277.74 mg/L) after 48 h when mannitol was used as the sole source of carbon. The pigment yield was also influenced by the types of carbon sources and peptones. As a result, RHP was demonstrated to be a suitable substrate for prodigiosin production. These results revealed that prodigiosin could be produced efficiently by S. marcescens using RHP.

Biotransformation of Tributyltin chloride by Pseudomonas stutzeri strain DN2

A bacterial isolate capable of utilizing tributyltin chloride (TBTCl) as sole carbon source was isolated from estuarine sediments of west coast of India and identified as Pseudomonas stutzeri based on biochemical tests and Fatty acid methyl ester (FAME) analysis. This isolate was designated as strain DN2. Although this bacterial isolate could resist up to 3 mM TBTCl level, it showed maximum growth at 2 mM TBTCl in mineral salt medium (MSM). Pseudomonas stutzeri DN2 exposed to 2 mM TBTCl revealed significant alteration in cell morphology as elongation and shrinkage in cell size along with roughness of cell surface. FTIR and NMR analysis of TBTCl degradation product extracted using chloroform and purified using column chromatography clearly revealed biotransformation of TBTCl into Dibutyltin dichloride (DBTCl2) through debutylation process. Therefore, Pseudomonas stutzeri strain DN2 may be used as a potential bacterial strain for bioremediation of TBTCl contaminated aquatic environmental sites.

Identification and characterization of rhizospheric microbial diversity by 16S ribosomal RNA gene sequencing

In the present study, samples of rhizosphere and root nodules were collected from different areas of Pakistan to isolate plant growth promoting rhizobacteria. Identification of bacterial isolates was made by 16S rRNA gene sequence analysis and taxonomical confirmation on EzTaxon Server. The identified bacterial strains were belonged to 5 genera i.e. Ensifer, Bacillus, Pseudomona, Leclercia and Rhizobium. Phylogenetic analysis inferred from 16S rRNA gene sequences showed the evolutionary relationship of bacterial strains with the respective genera. Based on phylogenetic analysis, some candidate novel species were also identified. The bacterial strains were also characterized for morphological, physiological, biochemical tests and glucose dehydrogenase (gdh) gene that involved in the phosphate solublization using cofactor pyrroloquinolone quinone (PQQ). Seven rhizoshperic and 3 root nodulating stains are positive for gdh gene. Furthermore, this study confirms a novel association between microbes and their hosts like field grown crops, leguminous and non-leguminous plants. It was concluded that a diverse group of bacterial population exist in the rhizosphere and root nodules that might be useful in evaluating the mechanisms behind plant microbial interactions and strains QAU-63 and QAU-68 have sequence similarity of 97 and 95% which might be declared as novel after further taxonomic characterization.

Bioinformatics based structural characterization of glucose dehydrogenase (gdh) gene and growth promoting activity of Leclercia sp. QAU-66

Glucose dehydrogenase (GDH; EC 1.1. 5.2) is the member of quinoproteins group that use the redox cofactor pyrroloquinoline quinoine, calcium ions and glucose as substrate for its activity. In present study, Leclercia sp. QAU-66, isolated from rhizosphere of Vigna mungo, was characterized for phosphate solubilization and the role of GDH in plant growth promotion of Phaseolus vulgaris. The strain QAU-66 had ability to solubilize phosphorus and significantly (p < 0.05) promoted the shoot and root lengths of Phaseolus vulgaris. The structural determination of GDH protein was carried out using bioinformatics tools like Pfam, InterProScan, I-TASSER and COFACTOR. These tools predicted the structural based functional homology of pyrroloquinoline quinone domains in GDH. GDH of Leclercia sp. QAU-66 is one of the main factor that involved in plant growth promotion and provides a solid background for further research in plant growth promoting activities.

Activation of the intrinsic mitochondrial apoptotic pathway in swine influenza virus-mediated cell death

The mitochondrial pathway of swine influenza virus (SIV)-induced apoptosis was investigated using porcine kidney (PK-15) cells, swine testicle (ST) cells, and HeLa cervical carcinoma cells which are known not to support viral replication. As judged by cell morphology, annexin V staining, and DNA fragmentation, PK-15 and ST cells infected with three different subtypes of SIV (H1N1, H3N2, and H1N2) were obviously killed by apoptosis, not necrosis. SIV infection in PK-15 and HeLa cells was shown to decrease the cellular levels of Bcl-2 protein compared to that of mock-infected control cells at 24 h post-infection, whereas expression levels of Bax protein increased in the PK-15 cells, but did not increase in HeLa cells by SIV infection. Cytochrome c upregulation was also observed in cytosolic fractions of the PK-15 and HeLa cells infected with SIV. Apoptosome (a multi-protein complex consisting of cytochrome c, Apaf-1, caspase-9, and ATP) formation was confirmed by immunoprecipitation using cytochrome c antibody. Furthermore, SIV infection increased the cellular levels of TAJ, an activator of the JNK-stressing pathway, and the c-Jun protein in the PK-15 and HeLa cells. Taken together, these results suggest that the mitochondrial pathway should be implicated in the apoptosis of PK-15 cells induced by SIV infection.

Purification and characterization of glutathione peroxidase from camel liver cytosol

Glutathione peroxidase [GSH: H2O2 oxidoreductase, EC 1.11.1.9] was purified to homogeneity from camel liver cytosol by ammonium sulfate fractionation and chromatography on CM-Sephadex C-50, DEAE-Sephadex A-50, followed by Sephadex G-200 and Sephadex G-100. The purified enzyme was homogeneous as judged by SDS-PAGE. The molecular weight of the enzyme as determined by SDS-PAGE was 66 +/- 2 kDa and that by gel filtration was comparable, indicated that the enzyme protein was a single polypeptide. The enzyme selenium content was calculated to be 4.4 g atoms/mole enzyme protein. The basic and acidic amino acids represent 40% of the total protein content. Optimum pH was 8.4 using tris-HCl buffer at 37C. Optimum temperature was 55C and the Km values for both GSH and cumene-OOH were 0.8 and 1.6 mM, respectively. Cadmium ions [II] was found to be a potent inhibitor. The purified enzyme was stable for more than 2 months under frozen conditions in the presence of dithiothreitol [DTT]

Cytosol cathepsin D in Thai women with breast cancer.

The necessity of screening high-risk patients with breast cancer who may require more intensive systemic therapy especially in the node negative subgroup was generally accepted. Cathepsin D, an estrogen induced protease, has been shown to be implicated in the proliferation and invasion of breast cancers. Retrospective assessment of cytosol cathepsin D in 151 primary breast cancers was done together with ER, PR and other clinico-pathological parameters. No significant relationship was shown between cathepsin D concentrations or cathepsin D status using median value of 56 pmol/mg protein as cutoff level with most studied parameters. High cathepsin D status was found in 47 per cent of patients with fibrocystic disease of breast and 30 per cent in node-negative, ER-PR negative tumors. Survival analysis after 5 or 10 year follow-up and evaluation in a larger scale are necessary before including cytosol cathepsin D measurement as a routine clinical investigation for breast cancers.

Optimización de la cuantificación de los receptores nucleares para estradiol y progesterona en útero de rata / Optimization in the quantification of nuclear receptors for estradiol and progesterona in uteri rat

La cuantificación de los receptores hormonales empleando hormona radiactiva en neoplasias hormonodependientes regularmente se realiza únicamente en el citosol celular. Esta determinacon es incompleta, ya que la presencia de los receptores localizados en el núcleo no se efectúa y, por lo tanto, no se determina la totalidad de los receptores en un tumor. En el presente estudio se otimizó la cuantificación de los receptores hormonales para estradiol localizados en el núcleo celular, empleando diferentes temperaturas de incubación con la hormona radiactiva (4,22 y 37 Grados C). Los receptores nucleares se extrajeron de la cromatina con soluciones amortiguadoras de KC1 0.4 M. se emplearon como modelo experimental úteros de ratas Wistar, adrenalectomizadas y ooforectomizadas, las cuales fueron posteriormente estimuladas con estradiol, progesterona y vehículo de administración. Se pudo observar que la cuantificación de los receptores nucleares para estradiol a 22 Grados C detecta un 13 por ciento más de estas proteínas, sin que se vea afectada por la temperatura la afinidad del receptor por su hormona. El receptor de progesterona no se pudo detectar en el núcleo, ya que al parecer es altamente sensible al KC1. En conclusión, es factible determinar los receptores nucleares para estradiol, empleando incubaciones a 22 Grados C como temperatura óptima en el núcleo.