Aqueous Two Phase System (ATPS) or liquid-liquid extraction is used in biotechnology to recover valuable compounds from raw sources. In Aqueous Two-Phase Systems, many factors influence the Partition coefficient, K, (which is the ratio of protein concentration in the top phase to that in the bottom phase) and the Recovery percentage (Rec%). In this research, two systems of ATPS were used: first, polyethylene glycol (PEG) 4000/Sodium citrate (SC), and the second, PEG8000/ Sodium phosphate (SPH), for the extraction of Bovine Serum Albumin (BSA). The behavior of Rec% and K of pure (BSA) in ATPS has been investigated throughout the study by the effects of five parameters: temperature, concentration of polyethylene glycol (PEG4000 and PEG8000), the concentration of Sodium citrate or Sodium phosphate, pH, and the addition of sodium chloride as a supporting agent. The recovery percentage of BSA and its partition coefficient are significantly influenced by these factors to various degrees. The most influential variable in this study is PEG concentration for both systems. In addition to the PEG concentration, the stabilizing impact of NaCl is a crucial factor. The interaction between biomolecules and PEG gets more hydrophobic as the PEG concentration is raised. In the first system (PEG4000/SC), the maximum recovery percentage and partition coefficient were 98.99% and 97.69, respectively, at 31°C, PEG4000 concentration 1.5g/10 ml, Sodium citrate concentration 2.7 g/10 ml, pH 10, and 0.5 M NaCl concentration. While in the second system (PEG8000/SPH), the maximum recovery percentage and partition coefficient was 98.93% and 92.12, respectively, at 31oC, PEG8000 concentration 1.5 g/10 ml, Sodium phosphate concentration 2.4 g/10 ml, pH 10, and concentration of NaCl 0.5 M.
Protein partitioning in an Aqueous Two-Phase System (ATPS) has been found to be a powerful method for extracting and separating mixtures of biomolecules. In Aqueous Two-Phase Systems, many factors influence the Partition coefficient (K) (which is the ratio of protein concentration in the top phase to that in the bottom phase) and the Recovery percent (Rec%). Two systems of ATPS were used: first, polyethylene glycol (PEG4000)/sodium phosphate (SPH), and second, PEG4000/Dextran. The behavior of Rec% and (K) of pure Bovine Serum Albumin (BSA) in (ATPS) has been investigated throughout the study of the effects of five parameters: (temperature (T), the concentration of polyethylene glycol (PEG4000), the concentration of sodium phosphate or Dextran, pH, and the addition of sodium chloride as a supporting agent). In both systems, pH was the more significant parameter on a Recovery percent (Rec%) and partition coefficient (K). In the first system, the maximum (Rec%) and (K) was 98.08% and 50.99, respectively, at a temperature of 31°C, the concentration of PEG4000 of 1.5 g/10 ml, the concentration of sodium phosphate at 2.4 g/10 ml, pH 10, and the concentration of NaCl at 0.5 M. While in the second system, the parameter that has a more significant effect on (Rec%) and (K) was the temperature. The maximum (Rec%) and (K) were 97.54% and 39.7 respectively at a temperature of 31°C, with a concentration of PEG4000 1.5 g/10 ml, the concentration of Dextran 2.4 g/10 ml, pH 5, and concentration of NaCl at 0.1 M.
Petroleum contamination in soil fields is a serious problem, particularly in countries that produce, transport, and refine oil. Phytoremediation is a promising approach for the cleanup of soil contaminated with petroleum hydrocarbons (PHCs) and heavy metals (HMs). For this purpose, this study was conducted to identify gene expression profiles for two related genes in response to petroleum stress in maize leaves at two contaminated sites. Two sites were chosen to collect polluted soil with oily residues: AL-Daura power station and AL-Daura oil refinery in Baghdad province. The soil was subjected to chronic pollution for many years. In addition, one unpolluted site was chosen for comparison as control around 4 Km upwind from ALDaura thermal power station. The gene expression of Opr1 and Pao1 genes in addition to ZmAct as reference gene was analysis in the maize leaves using RT-PCR. All data were statistically analyzed using analysis of variance, and treatment means were differentiated using the Least Significant Difference (LSD) test at P ≤ 0.05. The expression of Opr1 and Pao1 genes in maize leaves was overexpressing with 42.86 and 10.25 folds at refinery soil, respectively, while at power station soil increased with 5.16 and 5.04 folds, respectively but not differ significantly from control soil. The study show that the Real-time PCR analysis confirmed the expression patterns of the Opr1 and Pao1 genes, which play critical roles in mediating plant tolerance to petroleum stress.
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