Paper Information

Journal:   IRANIAN JOURNAL OF PUBLIC HEALTH   AUGUST 2014 , Volume 43 , Number SUPPLEMENT 2; Page(s) 50 To 50.
 
Paper: 

LIQUID–LIQUID EXTRACTION OF ALKALINE PROTEASES FROM FERMENTED BROTH BY PEG/SODIUM NITRATE AQUEOUS TWO-PHASE SYSTEM

 
 
Author(s):  KAMALI YAZDI KEIVAN, ANVARI MASOUMEH
 
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Abstract: 

Background: The objective of the study was to determine extraction conditions include polyethylene glycol (PEG) (%35, 40, 45), salt concentration (%25, 30, 35), phase volume ratio (%0.5, 0.75, 1) and system pH (8, 9, 10).
Methods: In this study, Bacillus sp. was isolated and identified by Kamali Yazdi and et al. The initial culture was grown in 250 ml Erlenmayer flask containing glucose (10 g/l), pepton (5 g/l), yeast extracct (5 g/l), Mgso4.7H2o (0.5% w/v) and the pH was adjusted (50 mM carbonate-bicarbonate buffer) to 9. The flasks were incubated at 37 C on rotation shaker at a speed of 250 rpm for 24 hours. PEG 6000/Sodium nitrate ATPS of different pH values (8, 9, 10) were prepared at room temperature. In a sterile test tube, 5.0 ml of PEG 6000, 5.0 ml of Sodium nitrate and 2.0 ml of enzyme from the fermented bacterial were taken and undisturbed for 10 minutes at room temperature. This mixture was centrifuged at 5, 000 rpm for 15 minutes. Top phase and bottom phase were collected and absorbance of sampels was measured at 280 nm.
Results: The best performance of the system was obtained applying 40 (%w/w) PEG 6000 concentration, 30 (%w/w) Sodium nitrate concentration and 1 (%w/w) phase volume ratio at pH 10 (119.88 U/ml).
Conclusion: Phase composition of the aqueos two-phase systems had a significant effect on enzyme partitioning. The PEG/Sodium nitrate system was proved suitable for alkaline protease removal from fermented broth of Bacillus sp. Four successive experimental designs were used in the present study to select the best conditions of protease extraction by PEG/Sodium nitrate ATPS.

 
Keyword(s): ALKALINE PROTEASE, AQUEOUS TWO-PHASE SYSTEM, LIQUID–LIQUID EXTRACTION, BACILLUS SP
 
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