title: Soluble expression and purification of the recombinant bioactive peptide precursor BPP-1 in Escherichia coli using a cELP-SUMO dual fusion system.
abstract: A bioactive peptide precursor (BPP-1, 14.3 kDa/115AA), a newly designed polypeptide that may exert a potential antihypertensive effect in vivo, is composed of many different ACE inhibitory peptides and antioxidant peptides tandemly linked according to the restriction sites of gastrointestinal proteases. In this report, we present a novel method to obtain soluble BPP-1 in Escherichia coli using cationic elastin-like polypeptide and SUMO (cELP-SUMO) tags. The cELP-SUMO-tagged fusion protein was expressed in soluble form at 20 °C for 20 h. After purification based on the inverse transition cycling (ITC) method, the purified cELP-SUMO-CFPP fusion protein was subsequently cleaved by a SUMO protease to release the mature BPP-1. After a subsequent simple salt precipitation process, approximately 167.2 mg of recombinant BPP-1 was obtained from 1 l of bacterial culture with at least 92% purity. The molecular mass (Mr) of the recombinant BPP-1 was confirmed by MALDI-TOF MS to equal 14,347. The purified BPP-1 was subjected to simulated gastrointestinal digestion, and the resulting hydrolysates exhibited notable ACE inhibitory and antioxidant activities in vitro. This report provides the first description of the soluble production of a bioactive peptide multimer with potential ACE inhibitory and antioxidant activities in E. coli using a cELP-SUMO tag.
authors.affiliation: School of Food Science and Engineering, Yangzhou University, Jiangsu, Yangzhou 225127, China. Electronic address: sqrao@yzu.edu.cn.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Rao
authors.affiliation: School of Food Science and Engineering, Yangzhou University, Jiangsu, Yangzhou 225127, China.
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Zang
authors.affiliation: School of Food Science and Engineering, Yangzhou University, Jiangsu, Yangzhou 225127, China.
authors.forename: Zhenquan
authors.initials: Z
authors.lastname: Yang
authors.affiliation: School of Food Science and Engineering, Yangzhou University, Jiangsu, Yangzhou 225127, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Gao
authors.affiliation: School of Food Science and Engineering, Yangzhou University, Jiangsu, Yangzhou 225127, China.
authors.forename: Yongqi
authors.initials: Y
authors.lastname: Yin
authors.affiliation: School of Food Science and Engineering, Yangzhou University, Jiangsu, Yangzhou 225127, China. Electronic address: wmfang@yzu.edu.cn.
authors.forename: Weiming
authors.initials: W
authors.lastname: Fang
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D010455:Peptides / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D025841:Small Ubiquitin-Related Modifier Proteins
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins
keywords: Activity; BPP-1; Escherichia coli; Soluble expression; cELP-SUMO