Background: Petroleum polycyclic aromatic hydrocarbons (PAHs) are known to be toxic and carcinogenic for humans and their contamination of soils and water is of great environmental concern. Identification of the key microorganisms that play a role in pollutant degradation processes is relevant to the development of optimal in situ bioremediation strategies. Objective: Detection of the ability of Pseudomonas fluorescens AH-40 to consume phenanthrene as a sole carbon source and determining the variation in the concentration of both nahAC and C23O catabolic genes during 15 days of the incubation period. Methods: In the current study, a bacterial strain AH-40 was isolated from crude oil polluted soil by enrichment technique in mineral basal salts (MBS) medium supplemented with phenanthrene (PAH) as a sole carbon and energy source. The isolated strain was genetically identified based on 16S rDNA sequence analysis. The degradation of PAHs by this strain was confirmed by HPLC analysis. The detection and quantification of naphthalene dioxygenase (nahAc) and catechol 2,3-dioxygenase (C23O) genes, which play a critical role during the mineralization of PAHs in the liquid bacterial culture were achieved by quantitative PCR. Results: Strain AH-40 was identified as pseudomonas fluorescens. It degraded 97% of 150 mg phenanthrene L-1 within 15 days, which is faster than previously reported pure cultures. The copy numbers of chromosomal encoding catabolic genes nahAc and C23O increased during the process of phenanthrene degradation. Conclusion: nahAc and C23O genes are the main marker genes for phenanthrene degradation by strain AH-40. P. fluorescence AH-40 could be recommended for bioremediation of phenanthrene contaminated site.

Several studies have reported the association of the SNP rs4077582 in the CYP11 gene with hyperandrogenism, which is one of the clinical manifestations of polycystic ovary syndrome (PCOS). These studies suggest that SNP rs4077582 may be involved in the etiopathogenisis of PCOS. To investigate whether the CYP11 gene SNP rs4077582 polymorphism is associated with the susceptibility to PCOS, we designed a case-controlled association study including 104 individuals. A case-controlled association study including 106 individuals (53 PCOS patients and 53 controls) was performed to assess the association of SNP rs4077582 with PCOS. Genotyping of SNP rs4077582 was conducted by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method that was performed on genomic DNA isolated from blood leucocytes. Results were analyzed in respect to clinical test results. The genotypic distributions of rs4077582 (CC, CT, TT) in the CYP11 gene in women with PCOS (CC, 29 (55.8%); CT, 20 (33.5%); and TT, 3 (5.8%). respectively) were significantly different from that in controls (CC, 44 (84.6%); CT, 6 (11.6%); and TT, 2 (3.8%) respectively) (P = 0.001). The allele frequencies in the PCOs women were: C, 78 (75%); and T, 26 (25%). This also was significantly different from the distribution in non-PCOs women: C, 94 (90.4%); and T, 10 (9.6%) (Table 2). In addition, rs4077582 C > T showed a association with PCOs by logistic regression analysis controlling for confounding factors. Our data suggest that SNP rs4077582 in the CYP11 gene is associated with susceptibility to PCOS.

Purpose: Strictosidine-β-D-glucosidase (SGD) is considered as a key enzyme in the productionof bisindole alkaloids in Catharanthus roseus. The present study illustrated the production of ashort sequence of this enzyme in Escherichia coli without codon optimization.Methods: Strictosidine-β-D-glucosidase (sgd) gene short sequence (1434 bp), which lacksthe conserved sequence KGFFVWS and the localization peptide sequence at the C-terminal,was amplified from cDNA of C. roseus leaves, cloned and expressed in Escherichia coli. Theactivity of the produced protein in cell free lysate was tested using total alkaloid extract of C.roseus leaves.Results: HPLC and LC-MS analysis of the assay mixture revealed the disappearance of thestrictosidine peak.Conclusion: SGD short sequence can be produced in Escherichia coli in active form withoutcodon optimization.