Expression and purification of antimicrobial peptide adenoregulin with C-amidated terminus in Escherichia coli

Cao, Wei; Zhou, Yuxun; Ma, Yushu; Luo, Qingping; Wei, Dongzhi

doi:10.1016/j.pep.2004.12.007

Cited by 41 publications

(30 citation statements)

References 24 publications

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“…In a previous study, culture and induction conditions were found to affect the expression of the soluble target proteins (Maldonado et al 2007). The highest level of soluble expression was achieved at 25°C and 0.4 mM IPTG with a yield reaching 23-28 % of total cell protein, which was higher than for other AMPs such as adenoregulin (Cao et al 2005), perinerin (Zhou et al 2007), and brevinin-2R (Mehrnejad et al 2008) (20, 20-25, and 25 %, respectively).…”

Section: Discussionmentioning

confidence: 91%

Molecular cloning and expression of ranalexin, a bioactive antimicrobial peptide from Rana catesbeiana in Escherichia coli and assessments of its biological activities

Aleinein

Hamoud

Schäfer

2012

Appl Microbiol Biotechnol

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The coding sequence, which corresponds to the mature antimicrobial peptide ranalexin from the frog Rana catesbeiana, was chemically synthesized with preferred codons for expression in Escherichia coli. It was cloned into the vector pET32c (+) to express a thioredoxin-ranalexin fusion protein which was produced in soluble form in E. coli BL21 (DE3) induced under optimized conditions. After two purification steps through affinity chromatography, about 1 mg of the recombinant ranalexin was obtained from 1 L of culture. Mass spectrometrical analysis of the purified recombinant ranalexin demonstrated its identity with ranalexin. The purified recombinant ranalexin is biologically active. It showed antibacterial activities similar to those of the native peptide against Staphylococcus aureus, Streptococcus pyogenes, E. coli, and multidrug-resistant strains of S. aureus with minimum inhibitory concentration values between 8 and 128 μg/ml. The recombinant ranalexin is also cytotoxic in HeLa and COS7 human cancer cells (IC50 = 13-15 μg/ml).

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Section: Discussionmentioning

confidence: 91%

Molecular cloning and expression of ranalexin, a bioactive antimicrobial peptide from Rana catesbeiana in Escherichia coli and assessments of its biological activities

Aleinein

Hamoud

Schäfer

2012

Appl Microbiol Biotechnol

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“…Several antibacterial peptides with biological activities have been expressed successfully by the E. coli expression system. According to the reports, Beta-defensin-4 from human [47,48], bovine LfcinB [36], porcine cecropin P1 [49], sarcotoxin IA from Sarcophaga peregrina larva [50] and adenoregulin from arboreal frog (Phyllomedusa bicolor) [51] were expressed with an E. coli expression system. Among aquatic species, previous reports have revealed that hepcidin from Tilapia (Oreochromis mossambicus) [52] and channel catfish [42], Piscidin-1 from Striped bass (Morone saxatilis) [53], LEAP-2 from Grass carp (Ctenopharyngodon idella) [54] have been successfully expressed via an E. coli expression system.…”

Section: Discussionmentioning

confidence: 99%

Expression, Purification and Antibacterial Activity of NK-Lysin Mature Peptides from the Channel Catfish (Ictalurus punctatus)

Cai

Wang

et al. 2016

Applied Sciences

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Antimicrobial peptides (AMPs) are small peptides and play important roles in host innate immune response against microbial invasion. Aquatic animals secrete different kinds of antimicrobial peptides which have antimicrobial activity towards microorganisms. NK-lysins, mature peptides produced by cytotoxic T lymphocytes and natural killer cells, are comprised of 74-78 amino acid residues, demonstrating broad-spectrum antimicrobial activity against bacteria, fungi, protozoa, and parasites. In this study, three distinct NK-lysin mature peptide (mNKLs), transcripts (76 amino acid residues) cloned from the channel catfish (Ictalurus punctatus) head kidney were ligated into plasmid vector pET-32a(+) to express the mNKLs fusion protein. The fusion protein was successfully expressed in E. coli Rosetta (DE3) under optimized conditions. After purification by affinity column chromatography, the fusion protein was successfully cleaved by enterokinase and released the peptide mNKLs. Tricine-SDS-PAGE results showed that mNKLs (approximately 8.6 kDa) were successfully expressed. The purified peptide mNKLs exhibited antibacterial activity against Staphylococcus aureus and E. coli.

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“…In our experiment, the soluble fusion GST-palustrin-2CE protein was expressed successfully, and the proportion reached 35%-39% of total cell proteins, higher than the antimicrobial peptide adenoregulin and the LH multimers (20% and 35% respectively). 27,28) In further studies, other strategies might be adopted to increase the activity of palustrin-2CE peptide.…”

Section: Discussionmentioning

confidence: 99%

Molecular Cloning, Expression, Purification, and Functional Characterization of Palustrin-2CE, an Antimicrobial Peptide ofRana chensinensis

Sun

et al. 2012

Bioscience, Biotechnology, and Biochemistry

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Expression and purification of antimicrobial peptide adenoregulin with C-amidated terminus in Escherichia coli

Cited by 41 publications

References 24 publications

Molecular cloning and expression of ranalexin, a bioactive antimicrobial peptide from Rana catesbeiana in Escherichia coli and assessments of its biological activities

Molecular cloning and expression of ranalexin, a bioactive antimicrobial peptide from Rana catesbeiana in Escherichia coli and assessments of its biological activities

Expression, Purification and Antibacterial Activity of NK-Lysin Mature Peptides from the Channel Catfish (Ictalurus punctatus)

Molecular Cloning, Expression, Purification, and Functional Characterization of Palustrin-2CE, an Antimicrobial Peptide ofRana chensinensis

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