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Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system

The potential use of recombinant collagen-like protein (recCLP) extracted from bacteria as disease-free collagen has been studied over the past decade. However, the complexity of the downstream processing generates high demand for an efficient and low-cost purification method. Aqueous two-phase syst...

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Main Authors: Nursyahidatul Azwa, Awang, Azura, Amid, Zatul Iffah, Mohd Arshad
Format: Article
Language:English
Published: International Islamic University Malaysia-IIUM 2023
Subjects:
QD Chemistry
T Technology (General)
TA Engineering (General). Civil engineering (General)
TP Chemical technology
Online Access:http://umpir.ump.edu.my/id/eprint/40637/1/Rhodopseudomonas%20palustris%20collagen-like%20recombinant.pdf
http://umpir.ump.edu.my/id/eprint/40637/
https://doi.org/10.31436/iiumej.v24i1.2468
https://doi.org/10.31436/iiumej.v24i1.2468
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spelling my.ump.umpir.406372024-04-30T06:39:11Z http://umpir.ump.edu.my/id/eprint/40637/ Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system Nursyahidatul Azwa, Awang Azura, Amid Zatul Iffah, Mohd Arshad QD Chemistry T Technology (General) TA Engineering (General). Civil engineering (General) TP Chemical technology The potential use of recombinant collagen-like protein (recCLP) extracted from bacteria as disease-free collagen has been studied over the past decade. However, the complexity of the downstream processing generates high demand for an efficient and low-cost purification method. Aqueous two-phase system (ATPS) was adopted as a new approach to the recovery of biomolecules due to its simple, benign, and straightforward process. This study aimed to purify recombinant collagen-like protein from Rhodopseudomonas palustris using ATPS formed by a polymer/salt system. Recombinant collagen-like protein from R. palustris was partitioned in ATPS composed of polyethylene glycol (PEG) and potassium phosphate and several factors that influence the protein partitioning such as volume ratio, system pH, the concentration of polymer and salt were studied. Then, optimization of the selected ATPS conditions (PEG and salt concentration) were performed using response surface methodology (RSM). Results showed that the optimum conditions were found in ATPS with 24.80% (w/w) PEG 2000 and 29.23% (w/w) potassium phosphate with recCLP concentration of 3.23 ± 0.12 mg/mL with purification factor 7.48 ± 0.3. In comparison with the affinity chromatography method, ATPS was found to be low-cost, and time-saving with a higher protein recovery. Hence, this study demonstrated the potential application of ATPS in the recovery of recombinant CLPs for large-scale downstream processing International Islamic University Malaysia-IIUM 2023 Article PeerReviewed pdf en cc_by_nc_4 http://umpir.ump.edu.my/id/eprint/40637/1/Rhodopseudomonas%20palustris%20collagen-like%20recombinant.pdf Nursyahidatul Azwa, Awang and Azura, Amid and Zatul Iffah, Mohd Arshad (2023) Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system. IIUM Engineering Journal, 24 (1). pp. 40-56. ISSN 1511-788X. (Published) https://doi.org/10.31436/iiumej.v24i1.2468 https://doi.org/10.31436/iiumej.v24i1.2468
institution Universiti Malaysia Pahang Al-Sultan Abdullah
building UMPSA Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Pahang Al-Sultan Abdullah
content_source UMPSA Institutional Repository
url_provider http://umpir.ump.edu.my/
language English
topic QD Chemistry
T Technology (General)
TA Engineering (General). Civil engineering (General)
TP Chemical technology
spellingShingle QD Chemistry
T Technology (General)
TA Engineering (General). Civil engineering (General)
TP Chemical technology
Nursyahidatul Azwa, Awang
Azura, Amid
Zatul Iffah, Mohd Arshad
Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
description The potential use of recombinant collagen-like protein (recCLP) extracted from bacteria as disease-free collagen has been studied over the past decade. However, the complexity of the downstream processing generates high demand for an efficient and low-cost purification method. Aqueous two-phase system (ATPS) was adopted as a new approach to the recovery of biomolecules due to its simple, benign, and straightforward process. This study aimed to purify recombinant collagen-like protein from Rhodopseudomonas palustris using ATPS formed by a polymer/salt system. Recombinant collagen-like protein from R. palustris was partitioned in ATPS composed of polyethylene glycol (PEG) and potassium phosphate and several factors that influence the protein partitioning such as volume ratio, system pH, the concentration of polymer and salt were studied. Then, optimization of the selected ATPS conditions (PEG and salt concentration) were performed using response surface methodology (RSM). Results showed that the optimum conditions were found in ATPS with 24.80% (w/w) PEG 2000 and 29.23% (w/w) potassium phosphate with recCLP concentration of 3.23 ± 0.12 mg/mL with purification factor 7.48 ± 0.3. In comparison with the affinity chromatography method, ATPS was found to be low-cost, and time-saving with a higher protein recovery. Hence, this study demonstrated the potential application of ATPS in the recovery of recombinant CLPs for large-scale downstream processing
format Article
author Nursyahidatul Azwa, Awang
Azura, Amid
Zatul Iffah, Mohd Arshad
author_facet Nursyahidatul Azwa, Awang
Azura, Amid
Zatul Iffah, Mohd Arshad
author_sort Nursyahidatul Azwa, Awang
title Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
title_short Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
title_full Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
title_fullStr Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
title_full_unstemmed Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
title_sort rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
publisher International Islamic University Malaysia-IIUM
publishDate 2023
url http://umpir.ump.edu.my/id/eprint/40637/1/Rhodopseudomonas%20palustris%20collagen-like%20recombinant.pdf
http://umpir.ump.edu.my/id/eprint/40637/
https://doi.org/10.31436/iiumej.v24i1.2468
https://doi.org/10.31436/iiumej.v24i1.2468
_version_ 1822924247829315584
score 13.244413

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