کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10156827 | 1666435 | 2018 | 35 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Optimization of in vitro refolding conditions of recombinant Lepidium draba peroxidase using design of experiments
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The main objective of this study was to optimize the in vitro refolding conditions of the recombinant Lepidium draba peroxidase (LDP). Initially, the effects of various factors were investigated on LDP refolding yield using one-factor-at-a-time (OFAT) method. Based on the OFAT results, optimum concentrations for LDP refolding were 2â¯M urea, 2â¯mM CaCl2, 0.42â¯mM l-glutathione oxidized (GSSG), 0.20â¯mg/ml protein, and 12â¯Î¼M hemin as well as pHâ¯7. Secondly, according to the OFAT results, design of experiments (DOE) was applied for investigation of the interactions between factors including protein (P), urea (U), CaCl2 (C), and GSSG (G). The results showed the possible interaction between PC, PG, PU, and GU. Lastly, response surface methodology (RSM) was used for final refolding conditions optimization. The final optimized refolding conditions for LDP were conducted as 2â¯M urea, 1â¯mM CaCl2, 0.70â¯mM GSSG, 0.07â¯mM DTT, 0.15â¯mg/ml protein which they obtained from RSM results and 12â¯Î¼M hemin, and pHâ¯7 according to the results of OFAT method. Overall, under optimal conditions, 23.4â¯mg active refolded LDP per liter of expression medium was obtained. So, the refolding yield was calculated to be approximately 48%.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 118, Part B, 15 October 2018, Pages 1369-1376
Journal: International Journal of Biological Macromolecules - Volume 118, Part B, 15 October 2018, Pages 1369-1376
نویسندگان
Yaser Fattahian, Ali Riahi-Madvar, Reza Mirzaee, Masoud Torkzadeh-Mahani, Gholamreza Asadikaram, Ghasem Sargazi,