Bacterial overexpression and purification of soluble recombinant human serum albumin using maltose-binding protein and protein disulphide isomerase

Minh Tan Nguyen, Yunseok Heo, Bich Hang Do, Sangki Baek, Chong Jai Kim, Yeon Jin Jang, Weontae Lee, Han Choe

Research output: Contribution to journalArticle

Abstract

Human serum albumin (HSA), the most abundant serum protein in healthy humans, plays important roles in many physiological processes and has wide clinical and research applications. Despite several efforts to obtain recombinant HSA (rHSA) from bacterial and eukaryotic expression systems, a low-cost and high-yield method for rHSA production is not available. The large molecular weight and high disulphide content hamper the expression and production of rHSA using bacterial hosts. Hence, a strategy that uses a fusion technique and engineered Escherichia coli strains was employed to improve the expression of soluble rHSA in the bacterial cytoplasm. The solubilities of the b'a' domain of human protein disulphide isomerase (PDIb'a')- and maltose-binding protein (MBP)-tagged rHSA expressed in Origami 2 at 18 °C were notably increased by up to 90.1% and 96%, respectively. A simple and efficient protocol for rHSA purification was established and approximately 9.46 mg rHSA was successfully obtained from a 500-mL culture at 97% purity. However, rHSA was mostly obtained in soluble oligomeric form. By introducing a simple refolding and size-exclusion chromatography step, monomeric rHSA was obtained at 34% yield. Native polyacrylamide gel electrophoresis confirmed the similarity in the molecular weights between E. coli-derived monomeric rHSA and commercial monomeric HSA.

Original languageEnglish
Article number105530
JournalProtein Expression and Purification
Volume167
DOIs
Publication statusPublished - 2020 Mar

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Maltose-Binding Proteins
Protein Disulfide-Isomerases
Serum Albumin
Molecular Weight
Physiological Phenomena
Escherichia coli
Native Polyacrylamide Gel Electrophoresis
Disulfides
Solubility
Gel Chromatography
Blood Proteins
Cytoplasm
Costs and Cost Analysis
Research

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Nguyen, Minh Tan ; Heo, Yunseok ; Do, Bich Hang ; Baek, Sangki ; Kim, Chong Jai ; Jang, Yeon Jin ; Lee, Weontae ; Choe, Han. / Bacterial overexpression and purification of soluble recombinant human serum albumin using maltose-binding protein and protein disulphide isomerase. In: Protein Expression and Purification. 2020 ; Vol. 167.
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Bacterial overexpression and purification of soluble recombinant human serum albumin using maltose-binding protein and protein disulphide isomerase. / Nguyen, Minh Tan; Heo, Yunseok; Do, Bich Hang; Baek, Sangki; Kim, Chong Jai; Jang, Yeon Jin; Lee, Weontae; Choe, Han.

In: Protein Expression and Purification, Vol. 167, 105530, 03.2020.

Research output: Contribution to journalArticle

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T1 - Bacterial overexpression and purification of soluble recombinant human serum albumin using maltose-binding protein and protein disulphide isomerase

AU - Nguyen, Minh Tan

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AU - Kim, Chong Jai

AU - Jang, Yeon Jin

AU - Lee, Weontae

AU - Choe, Han

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