Cloning, expression and purification of escherichia coli modified phytase

Introduction: Phytases are the class of phosphatases, which are capable of hydrolyzing phytic acid. Phytases with the phytate degradation are able to reduce or eliminate the harmful effects of phytate. Acidic and thermal stable phytase with high yield and purity by a relatively inexpensive system had extensive application. So, in this study, by modification in enzyme sequence, recombinant phytase production with the shorter length and high expression level was assessed. Materials and Methods: The phytase gene sequence was obtained from the NCBI database. After bioinformatics studies and doing the noted modification for increasing protein expression, gene proliferation was done by using PCR. E. coli BL21 (DE3) was used to express the protein. Protein purification was performed by Ni-NTA kit and finally, enzyme activity was assessed. Results: Phytase was successfully expressed and purified. Enzyme activity assay showed a significant activity. Conclusion: Produced recombinant phytase had high activity in spite of eliminating parts of the enzyme. © 2017, Semnan University of Medical Sciences. All rights reserved

Common bile duct stone associated with hemolytic uremic syndrome

Cholelithiasis is an unusual complication of hemolytic uremic syndrome. A 12-year-old boy with hemolytic uremic syndrome, established by renal biopsy, who developed cholestatic jaundice is presented here. Laboratory results for secondary causes of hemolytic uremic syndrome were normal. Abdominal ultrasonography and magnetic resonance cholangiopancreatography revealed extrahepatic obstruction. A common bile duct stone, discovered by retrograde cholanglopancreatography was extracted by sphincterotomy. In conclusion, cholelithiasis should be considered as a cause of abdominal pain and cholestasis in patients who are diagnosed as having hemolytic uremic syndrome

Effect of number of planting rows in strip intercropping of maize, pintobean and soybean and their sole cropping on seed yield

To evaluate the effects of rows of maize-pintobean, maize-soybean planted in strip intercropping and their sole croppings, an experiment using randomized complete block design with three replication, was conducted for two consecutive years at the Agricultural Research Center of Islamic Azad University, Tabriz branch. Treatments were: A (intercropping of maize-pintobean, one row of each, B (intercropping of maize-pintobean, two rows of each), C (intercropping of maize-pintobean, three rows of each), D (intercropping of maize-soybean, two rows of each), E (intercropping of maize-soybean, two rows of each), F (intercropping of maize-soybean, three rows of each), G(sole cropping of maize), H (sole cropping of pintobean) and I (sole cropping of soybean). Experimental results showed that maize produced, in both years, highest (17818 kg/ha) using treatment D and lowest (15509 kg/ha) yields in treatment G. This indicates that maize produced higher yield in intercropping than sole cropping. Maize, also, produced highest yield in treatment D and lowest in treatment F. Pintobean had highest yield (2498 kg/ha) in treatment H and lowest (2073 kg/ha) in treatment A. Soybean, on the other hand, produced highest yield (4794 kg/ha) in treatment I and also lowest (3257 kg/ha) in treatment D. A a whole, it may be concluded that pintobean and soybean produce higher yield in sole cropping as compared to intercroppings

Cloning and High-Level Expression of the Enzymatic Region of Phytase in E. coli

Phytase is an important enzyme poses great nutritional significance in humans and monogastric animals diets. The phytase production yield using wild sources, including micro-organisms, plants, and animals is sorely low. Thus, recombinant expression of phytase has received increasing interest for achieving production rate. Escherichia coli is the most preferred host for expression of heterologous proteins but overexpression of recombinant phytase in E. coli, met with limited success due to the sequestration of the enzyme into inclusion bodies. In the present study, artificial phytases gene with excellent thermostability and activity were designed by detecting the enzymatic region of the E. coli phytase gene by employing bioinformatics tools. Then, the PCR amplified recombinant gene was expressed in E. coli and the active enzyme was recovered from inclusion bodies. Employing cysteine amino acid in the dialysis buffer succeed to the superior activity of the enzyme with a specific activity of 73.8 U/mg. The optimum temperature and pH for enzyme activity were determined at 60 °C and 4, respectively. The novel recombinant enzyme illustrated perfect thermostability up to 70 °C with maintenance 75 of its activity. The enzyme was stable at pH range of 2�10. Moreover, the effects of ions and chemical compounds on enzyme stability and activity were assessed. © 2019, Springer Nature B.V

The Comparison Of Anthropometric and Physiologic Characteristics Between Isfahan juniors Football Players

Abstract This research has been performed on 34 volunteer male basketball players aged 12-14 age groups from junior male basketball team of Ankara University and Ulaútırma Sports Club. The subjects have been randomly divided into the experimental and control groups . Before training program resting heart rate , systolic-diastolic blood pressure, aerobic-anaerobic capacity have been evaluated for both groups, besides some measurements have been also performed on some parameters by using Eurofit test battery. The same measurements have been performed again after the 8 weeks combined training program for the experimental group and the effect of combined trainings on these parameters has been tried to determine. In this 8 weeks period, the subjects in the control group have not been put to an extra training program, they have carried on their daily sports life. SPSS 8.0 statistical package program has been used for the statistical analysis and arithmetic average, standart deviation, standart error and range of the values have been determined. The differences between groups have been examined by t-test in the significance level 0.01 and 0.05. In consequence of this research, it has been observed that while the values of body height (% 0,61 ; p <.001), leg power (% 17,21 ; p <.001), pull-up motion (% 32,06 ; p < .001), standing long jump (% 6,85 ; p < .001) and aerobic capacity (% 8,65 ; p < .001) have showed an increase, the values of resting heart rates (% 7,71; p < .005), 10 x 5 m. (% 13,15 ; p < . 001) and left hand reaction time to light (% 7,31; p < .0005) have showed a decrease after the first and the last measurements of combined training program for the experimental group. It has not been observed any changes in the control group. As a result, it has been determined that the combined trainings performed on experimental group have been more efficient than technical trainings