PMID- 29959013
OWN - NLM
STAT- MEDLINE
DCOM- 20181121
LR  - 20181121
IS  - 1879-0003 (Electronic)
IS  - 0141-8130 (Linking)
VI  - 118
IP  - Pt B
DP  - 2018 Oct 15
TI  - Optimization of in vitro refolding conditions of recombinant Lepidium draba 
      peroxidase using design of experiments.
PG  - 1369-1376
LID - S0141-8130(18)31832-4 [pii]
LID - 10.1016/j.ijbiomac.2018.06.122 [doi]
AB  - 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 2 M urea, 2 mM CaCl(2), 0.42 mM 
      l-glutathione oxidized (GSSG), 0.20 mg/ml protein, and 12 muM hemin as well as 
      pH 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), CaCl(2) (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 2 M urea, 1 mM CaCl(2), 0.70 mM 
      GSSG, 0.07 mM DTT, 0.15 mg/ml protein which they obtained from RSM results and 
      12 muM hemin, and pH 7 according to the results of OFAT method. Overall, under 
      optimal conditions, 23.4 mg active refolded LDP per liter of expression medium 
      was obtained. So, the refolding yield was calculated to be approximately 48%.
CI  - Copyright (c) 2018 Elsevier B.V. All rights reserved.
FAU - Fattahian, Yaser
AU  - Fattahian Y
AD  - Department of Biotechnology, Institute of Science and High Technology and 
      Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
FAU - Riahi-Madvar, Ali
AU  - Riahi-Madvar A
AD  - Department of Biotechnology, Institute of Science and High Technology and 
      Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran. 
      Electronic address: Riahi.ali@gmail.com.
FAU - Mirzaee, Reza
AU  - Mirzaee R
AD  - Jiroft University of Medical Sciences, Jiroft, Iran.
FAU - Torkzadeh-Mahani, Masoud
AU  - Torkzadeh-Mahani M
AD  - Department of Biotechnology, Institute of Science and High Technology and 
      Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
FAU - Asadikaram, Gholamreza
AU  - Asadikaram G
AD  - Neuroscience Research Center, Institute of Neuropharmacology, Department of 
      Biochemistry, School of Medicine, Kerman University of Medical Sciences, Kerman, 
      Iran.
FAU - Sargazi, Ghasem
AU  - Sargazi G
AD  - Department of Nanotechnology Engineering, Mineral Industries Research Center, 
      Shahid Bahonar University of Kerman, Kerman, Iran.
LA  - eng
PT  - Journal Article
DEP - 20180626
PL  - Netherlands
TA  - Int J Biol Macromol
JT  - International journal of biological macromolecules
JID - 7909578
RN  - 0 (Recombinant Proteins)
RN  - 8W8T17847W (Urea)
RN  - EC 1.11.1.7 (Peroxidase)
RN  - M4I0D6VV5M (Calcium Chloride)
RN  - ULW86O013H (Glutathione Disulfide)
SB  - IM
MH  - Calcium Chloride/pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Glutathione Disulfide/pharmacology
MH  - Lepidium/*enzymology
MH  - Peroxidase/*chemistry
MH  - Protein Refolding/*drug effects
MH  - Recombinant Proteins/*chemistry
MH  - Solubility
MH  - Urea/pharmacology
OTO - NOTNLM
OT  - Design of experiments
OT  - Lepidium draba peroxidase
OT  - Refolding
OT  - Response surface methodology
EDAT- 2018/07/01 06:00
MHDA- 2018/11/22 06:00
CRDT- 2018/07/01 06:00
PHST- 2018/04/18 00:00 [received]
PHST- 2018/06/17 00:00 [revised]
PHST- 2018/06/25 00:00 [accepted]
PHST- 2018/07/01 06:00 [pubmed]
PHST- 2018/11/22 06:00 [medline]
PHST- 2018/07/01 06:00 [entrez]
AID - S0141-8130(18)31832-4 [pii]
AID - 10.1016/j.ijbiomac.2018.06.122 [doi]
PST - ppublish
SO  - Int J Biol Macromol. 2018 Oct 15;118(Pt B):1369-1376. doi: 
      10.1016/j.ijbiomac.2018.06.122. Epub 2018 Jun 26.