1. Cloning, expression, and purification of porcine adrenocorticotropic hormone in Escherichia coli
- Author
-
Feng Jun, Meiqing Feng, Huang Zongqing, Haoju Hua, and Wu Yong
- Subjects
0106 biological sciences ,Swine ,medicine.medical_treatment ,Recombinant Fusion Proteins ,lac operon ,Gene Expression ,Adrenocorticotropic hormone ,medicine.disease_cause ,01 natural sciences ,law.invention ,03 medical and health sciences ,Adrenocorticotropic Hormone ,law ,010608 biotechnology ,medicine ,Escherichia coli ,Animals ,Cloning, Molecular ,030304 developmental biology ,Cloning ,0303 health sciences ,Protease ,Molecular mass ,Chemistry ,Fusion protein ,Biochemistry ,Recombinant DNA ,Biotechnology - Abstract
Adrenocorticotropic hormone (ACTH) is an old medicine derived from porcine pituitary gland that has been marketed for more than 60 years. In this study, we present a recombinant approach to produce ACTH in Escherichia coli (E. coli). The SUMO-tagged fusion protein was cloned and expressed after induction with isopropyl-β- d -thiogalactopyranoside (IPTG) at 25 °C for 8 h. The fusion protein was extracted and purified by anion exchange chromatography, and the SUMO tag was subsequently removed by digestion with ubiquitin-like protease 1 (ULP1). Approximately 95.3 mg of recombinant ACTH with 94.2% purity was obtained after cation exchange purification performed on a 5 mL column, from 286 mL fermentation broth based on the amount of pellets homogenized. The molecular mass of the recombinant ACTH was confirmed by mass spectrometry to equal 4567.32 Da.
- Published
- 2020