Fusion expression of bovine lactoferricin in Escherichia coli

Feng, Xingjun; Wang, Jianhua; Shan, Anshan; Teng, Da; Yang, Yalin; Yao, Yi Ju; Yang, Guanpin; Shao, Yanchun; Liu, Shuo; Fan, Zhang

doi:10.1016/j.pep.2005.08.016

Cited by 51 publications

(25 citation statements)

References 30 publications

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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%

“…The pMD19-T-mNKLs were digested with restriction enzymes and then cloned into the pET-32a(+) plasmid vector, which was also confirmed via restriction digest and sequencing ( Figure 2). [36]. Regions of identity (*), strong similarity (:) and weak similarity (.)…”

Section: Cloning and Construction Of Recombinant Expression Vectormentioning

confidence: 99%

“…In order to obtain the recombinant mNKLs with similar structure of native NK-lysins, the enterokinase cleavage site (DDDDK) and termination codon (TGA) were added in N-terminal and C-terminal of mNKLs, respectively; (C) Fusion protein produced as translation of pET-32a(+) vector. [36]. Regions of identity (*), strong similarity (:) and weak similarity (.)…”

Section: Cloning and Construction Of Recombinant Expression Vectormentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Cloning and Construction Of Recombinant Expression Vectormentioning

confidence: 99%

Section: Cloning and Construction Of Recombinant Expression Vectormentioning

confidence: 99%

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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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“…Luo et al (2007) have expressed recombinant bovine LF N-terminal peptide in E.coli, and the product displayed obvious antimicrobial activity. Bovine LF derivates lactoferricin (LfcinB; Feng et al 2006), LfcinB15 (Tian et al 2007) and lactoferrampin (LfampinB;Yang et al 2009) were expressed in E. coli system and displayed obvious antibacterial activity. Choi et al (2008) expressed recombinant human lactoferrin in glycoengineered Pichia pastoris to evaluate the effects of terminal Nglycan structures on immune responses.…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and characterization of Kunming mice lactoferrin and its N-lobe

Wang¹,

Tian²,

Teng³

et al. 2010

Biometals

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The lactoferrin cDNA of Kunming mice was isolated by reverse transcription polymerase chain reaction and cloned into vector pET28a(+). Its deduced amino acid sequence was analyzed and compared with lactoferrin of other species. Its secondary and tertiary structure are predicted and modeled by bioinformatics tools online. Then recombinant Kunming mice lactoferrin and its N-lobe were both expressed successfully in the Escherichia coli BL21(DE3) in the form of inclusion bodies. After purification with Ni-NTA His-Bind resin, the yield of recombinant lactoferrin was 17 mg l(-1) with purity of 92.1%, and that of lactoferrin N-lobe was 20 mg l(-1) with purity of 98.5%. The inhibition efficiency of refolded lactoferrin N-lobe against Staphylococcus aureus ATCC 25923 reaches 48.6% at the concentration of 25 micromol l(-1). However, the refolded lactoferrin (12.5 micromol l(-1)) didn't display obvious inhibition activity in the test. The expression of recombinant Kunming mice lactoferrin and its N-lobe will be helpful for the study of lactoferrin on structure, function and application in a mouse model system.

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“…For example, a strategy by improving the thrombin cleavage rate for high-level expression of human interferon α (IFNα) as the GST fusion protein in E. coli was referred [17]. The recombinant bovine lactoferricin (LfcinB) was expressed as the GST fusion protein, which contains a Factor Xa cleavage site, and a thrombin cleavage site, but functional LfcinB was only cleaved by thrombin protease, not by Factor Xa [18].…”

Section: Introductionmentioning

confidence: 99%

Enhancement of Recombinant Human ADAM15 Disintegrin Domain Expression Level by Releasing the Rare Codons and Amino Acids Restriction

Lian-fen

Lei

et al. 2008

Appl Biochem Biotechnol

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This study was aimed at increasing the production of the recombinant human ADAM15 disintegrin domain (RADD) in Escherichia coli by releasing the rare codons and restricting amino acid residues. Three different strategies for increasing RADD production were examined: to select the suitable host strain, to optimize the rare codons, and to delete the amino acids residues. The total fusion protein glutathione-S-transferase (GST)-RADD concentration of 298 mg/l and 326 mg/l were achieved by selecting E. coli Rosetta (DE3) as the host strain and by changing GGA to GGC at the GST-RADD cassette, respectively. The RADD concentration was increased by 35.7% by eliminating the "Pro-Glu-Phe" residues at the GST-RADD junction. By combinational utilizing the preferred codon introduction and amino acid sequence optimization in E. coli Rosetta (DE3), the highest RADD concentration of 68 mg/l was achieved. The proposed strategy may provide an alternative approach for other enhanced recombinant protein production by E. coli.

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Fusion expression of bovine lactoferricin in Escherichia coli

Cited by 51 publications

References 30 publications

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

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

Cloning, expression and characterization of Kunming mice lactoferrin and its N-lobe

Enhancement of Recombinant Human ADAM15 Disintegrin Domain Expression Level by Releasing the Rare Codons and Amino Acids Restriction

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