High-Level Expression of a Soluble Functional Antimicrobial Peptide, Human β-Defensin 2, in Escherichia coli

Xu, Zhinan; Li, Peng; Zhong, Zhixia; Fang, Xiangming; Chen, Peilin

doi:10.1021/bp0502680

Cited by 38 publications

(29 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast to earlier reports about defensin production in E. coli K-12 strains (Huang et al, 2006;Pazgier and Lubkowski, 2006;Xu et al, 2006), this study reports for the first time the production of active HBD2 defensin in a probiotic E. coli strain (Nissle 1917), which is licensed as a drug for the treatment of patients suffering from e.g. diarrhea or ulcerative colitis.…”

Section: Listeria Monocytogenescontrasting

confidence: 59%

See 1 more Smart Citation

Construction of recombinant E. coli Nissle 1917 (EcN) strains for the expression and secretion of defensins

Seo¹,

Weibel

Wehkamp

et al. 2012

International Journal of Medical Microbiology

View full text Add to dashboard Cite

Section: Listeria Monocytogenescontrasting

confidence: 59%

“…It has been reported that a fusion expression system with thioredoxin (TrxA) resulted in an increased yield of soluble products (LaVallie et al 2000). This has been observed for HBD2 and HBD3 (Huang et al, 2006;Xu et al, 2006). …”

supporting

confidence: 67%

Construction of recombinant E. coli Nissle 1917 (EcN) strains for the expression and secretion of defensins

Seo¹,

Weibel

Wehkamp

et al. 2012

International Journal of Medical Microbiology

View full text Add to dashboard Cite

“…In addition to the cultivation temperature, the type of fusion protein utilized affected the yield of soluble HPV16 L1 production in E. coli. Our observation of the relatively lower yield of soluble HPV16 L1 upon fusion with GST is consistent with the previous findings of other groups, who reported a soluble GST-HPV16 L1 yields of less than 50% in E. coli [6,20]. Consistent with our findings, the MBP fusion tag was reported to be a more efficient solubilizing partner than GST for a set of proteins [21,22].…”

Section: Discussionsupporting

confidence: 93%

Expression, Purification, and Antibody Binding Activity of Human Papillomavirus 16 L1 Protein Fused to Maltose Binding Protein

Cho

Hahm²,

Kim³

et al. 2007

PPL

View full text Add to dashboard Cite

Genetic human papillomavirus type 16 L1 (HPV16 L1) has been widely studied for cervical cancer vaccine development. For the enzyme-linked immunosorbent assay (ELISA) screening of these vaccines, HPV16 L1 protein, which is required as a coating protein, has previously been expressed from costly and laborious recombinant baculovirus-infected insect cells. For a novel HPV16 L1 expression system characterized by a high yield of soluble form with simple purification steps, we have cloned and expressed two different types of HPV16 L1, both fused to maltose binding protein (MBP) or glutathione-S-transferase (GST) in Escherichia coli. The yield of soluble HPV16 L1 was influenced by the cultivation temperature. The yield of soluble form in the total MBP-fused HPV16 L1 protein (MBP-HPV16 L1) was 35% at 37 degrees C, but increased to 85% at 22 degrees C. Among the fusion partners, MBP provided higher yields of total and soluble HPV16 L1 than did GST. MBP-HPV16 L1 showed a 4.9-fold higher yield of the soluble form over insoluble inclusion bodies under optimized culture conditions. The soluble form of MBP-HPV16 L1 was purified via MBP affinity chromatography in a recovery yield of 9.7%. After fusion with MBP, HPV16 L1 showed binding activity to HPV16 L1-specific monoclonal antibody comparable to HPV16 L1 from the insect cells in ELISA tests. These results demonstrate that the use of MBP as a fusion partner may generate a high yield of soluble HPV16 L1 under optimized temperature conditions, and that MBP-fused HPV16 L1 might be applied further in evaluations of the immune responses of HPV16 L1-based cervical cancer vaccines.

show abstract

“…HBD-1 was cleaved from the fusion by protease 4 and the peptide was found to be active. Chen and colleagues have been improving the expression of HBD-2 for years [82,83]. They found that tandem expression and codon optimization could improve the expression level of the peptide.…”

Section: Peptide Productionmentioning

confidence: 99%

Tool Developments for Structure-Function Studies of Host Defense Peptides

Wang¹

2007

PPL

View full text Add to dashboard Cite

Antimicrobial peptides, or host defense peptides, are universal signaling and effector molecules in host defense and innate immunity. This article highlights various tools developed for cathelicidins and defensins, ranging from peptide identification, production, and structural biology, including the eight databases for antimicrobial peptides. Novel peptides can be identified from natural sources at both gene and protein levels. Solid-phase synthesis and bacterial expression are the two important methods for peptide production. Three-dimensional structures of antimicrobial peptides, primarily determined by solution NMR techniques, are essential for an in-depth understanding of the mode of action. The introduction of octanoyl phosphatidylglycerol as a bacterial membrane-mimetic model provides new insights into peptide-lipid interactions. The incorporation of structure and activity data into the antimicrobial peptide database (http://aps.unmc.edu/AP/main.html) will lead to an integrated understanding of these peptides via structural bioinformatics.

show abstract

High-Level Expression of a Soluble Functional Antimicrobial Peptide, Human β-Defensin 2, in Escherichia coli

Cited by 38 publications

References 20 publications

Construction of recombinant E. coli Nissle 1917 (EcN) strains for the expression and secretion of defensins

Construction of recombinant E. coli Nissle 1917 (EcN) strains for the expression and secretion of defensins

Expression, Purification, and Antibody Binding Activity of Human Papillomavirus 16 L1 Protein Fused to Maltose Binding Protein

Tool Developments for Structure-Function Studies of Host Defense Peptides

Contact Info

Product

Resources

About