Expression and purification of antimicrobial peptide buforin IIb in Escherichia coli

Abstract

A novel production method in Escherichia coli for an antimicrobial peptide of 21 amino acids, buforin IIb, which is a synthetic analog of buforin II, has been developed. The buforin IIb gene was cloned into the vector pET32a to construct an expression vector pET32a-buforin IIb. The fusion protein Trx-buforin IIb, purified by nickel nitrilo-triacetic acid (Ni-NTA) resin chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant buforin IIb. Purification of recombinant buforin IIb was achieved by HPLC: about 3.1 mg/l active recombinant buforin IIb with purity >99% was obtained. The recombinant buforin IIb showed antimicrobial activities that were similar to the synthetic one.

Antimicrobial peptides as natural bio-preservative to enhance the shelf-life of food

“…Several peptides are synthesized by this method. Generally, E. coli is used as a host organism for the synthesis of peptides (Sewald and Jakubke 2002;Wang et al 2011). Among all the above methods for antimicrobial peptides synthesis, recombination technique is better as compared to chemical and enzymatic methods.…”

Current scenario of peptide-based drugs: the key roles of cationic antitumor and antiviral peptides

“…Many groups have been performing recombinant technology in order to optimize the production yield of cationic peptides using animal cells (Brocal et al, 2006), yeast (Wang et al, 2009b), plant (Lee et al, 2011), and bacteria systems (Wang et al, 2011). …”

“…This bacterium has been used for expression of cecropin (Liang et al, 2006), lactoferricin (Luo et al, 2007), human α and β defensins (Wang et al, 2010b), buforin (Wang et al, 2011), indolicin (Morin et al, 2006), and LL-37 (Moon et al, 2006). Two different systems for expression of cecropin, one fused to enterokinase (Xu et al, 2007a) and other hybrid system fused to ubiquitin (Xu et al, 2007b) were constructed, and both systems were active against Gram-positive and negative bacteria, and fungi.…”

Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system

“…In the present study, novel hybrid protein combining PI with hLY was designed. Considering codon optimization facilitates the recombinant expression of many heterogeneous proteins or AMPs derived from various sources [24,40,41], we firstly optimized the coding sequence of rehLY-PI with biased codons of P. pastoris and successfully constructed the P. pastoris rehLY-PI/pPICZaA transformant. The production of recombinant rehLY-PI reached 86 ± 12.5 mg/L after 96 h induction with 0.5% methanol in a shaking flask.…”