SUMO-based expression and purification of dermcidin-derived DCD-1L, a human antimicrobial peptide, in<i>Escherichia coli</i>

Mohanraj, Ushanandini; Kinnunen, Oona; Kaya, Meryem Ecem; Aranko, Aino Sesilija; Viskari, Heli

doi:10.1101/343418

Cited by 3 publications

(2 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…, , ; Mohanraj et al . ), indicating a preference of the SUMO‐based system for recombinant production of the AMPs. Moreover, the combination of pET30a and ZYM‐5052 medium amplified the yield of AP2 in our study.…”

Section: Resultsmentioning

confidence: 99%

Expression and purification of antimicrobial peptide AP2 using SUMO fusion partner technology inEscherichia coli

Zhou

et al. 2018

Lett Appl Microbiol

View full text Add to dashboard Cite

Significance and Impact of the Study: AP2, an artificial mutant apidaecin (AP) peptide based on APs, has improved activities and may be regarded as a promising antibiotic alternatives. The secreted expression of antimicrobial peptide is of the greatest challenges because the antibacterial activity is not beneficial to the host. Our data suggest that the recombinant fusion strategy allows convenient high yield and easy purification of recombinant AP2. AbstractApidaecins (APs) are proline-rich antimicrobial peptides that were isolated from Apismelifera. APs possess broad-spectrum activities against Gram-negative bacteria and exhibit immune-modulatory functions. AP2, an artificial mutant AP peptide with improved activities, was expressed in Escherichia coli expression system using small ubiquitin-related modifier (SUMO) fusion technology and ZYM-5052 auto-induction medium. Approximately 23 mg of recombinant fusion protein smt3AP2 was purified per litre cultivated medium. After SUMO protease (Ulp) cleavage of smt3AP2, recombinant AP2 was further purified by affinity and cation exchange chromatography. The pure recombinant AP2 with calculated value of 2Á23 kDa reached a yield of 2Á7 mg l À1 and exhibited powerful antibacterial activity towards E. coli K88 with minimum inhibitory concentration at 5 lg ml À1 . The recombinant fusion strategy presented in this study allows convenient high yield and easy purification of recombinant AP2.

show abstract

Section: Resultsmentioning

confidence: 99%

Expression and purification of antimicrobial peptide AP2 using SUMO fusion partner technology inEscherichia coli

Zhou

et al. 2018

Lett Appl Microbiol

View full text Add to dashboard Cite

show abstract

“…On the other hand, production of recombinant LsGRP1 C protein was assisted with the help of yeast SUMO tag, in E. coli host system, which led to a high yield of SUMO-LsGRP1 C soluble fusion protein [28]. Likewise, SUMO-based DCD-1L production in E. coli was also reported recently [29].…”

Section: Biotechnological Approaches For Enhanced Antimicrobial Peptimentioning

confidence: 99%

Antimicrobial Peptides: Recent Insights on Biotechnological Interventions and Future Perspectives

Sinha¹,

Shukla

2019

PPL

View full text Add to dashboard Cite

With the unprecedented rise of drug-resistant pathogens, particularly antibiotic-resistant bacteria, and no new antibiotics in the pipeline over the last three decades, the issue of antimicrobial resistance has emerged as a critical public health threat. Antimicrobial Peptides (AMP) have garnered interest as a viable solution to this grave issue and are being explored for their potential antimicrobial applications. Given their low bioavailability in nature, tailoring new AMPs or strategizing approaches for increasing the yield of AMPs, therefore, becomes pertinent. </P><P> The present review focuses on biotechnological interventions directed towards enhanced AMP synthesis and revisits existing genetic engineering and synthetic biology strategies for production of AMPs. This review further underscores the importance and potential applications of advanced gene editing technologies for the synthesis of novel AMPs in future.

show abstract

A Toxicity Screening Approach to Identify Bacteriophage-Encoded Anti-Microbial Proteins

Mohanraj

Wan

Spruit

et al. 2019

Viruses

Self Cite

View full text Add to dashboard Cite

The rapid emergence of antibiotic resistance among many pathogenic bacteria has created a profound need to discover new alternatives to antibiotics. Bacteriophages, the viruses of microbes, express special proteins to overtake the metabolism of the bacterial host they infect, the best known of which are involved in bacterial lysis. However, the functions of majority of bacteriophage encoded gene products are not known, i.e., they represent the hypothetical proteins of unknown function (HPUFs). In the current study we present a phage genomics-based screening approach to identify phage HPUFs with antibacterial activity with a long-term goal to use them as leads to find unknown targets to develop novel antibacterial compounds. The screening assay is based on the inhibition of bacterial growth when a toxic gene is expression-cloned into a plasmid vector. It utilizes an optimized plating assay producing a significant difference in the number of transformants after ligation of the toxic and non-toxic genes into a cloning vector. The screening assay was first tested and optimized using several known toxic and non-toxic genes. Then, it was applied to screen 94 HPUFs of bacteriophage φR1-RT, and identified four HPUFs that were toxic to Escherichia coli. This optimized assay is in principle useful in the search for bactericidal proteins of any phage, and also opens new possibilities to understanding the strategies bacteriophages use to overtake bacterial hosts.

show abstract

SUMO-based expression and purification of dermcidin-derived DCD-1L, a human antimicrobial peptide, inEscherichia coli

Cited by 3 publications

References 23 publications

Expression and purification of antimicrobial peptide AP2 using SUMO fusion partner technology inEscherichia coli

Expression and purification of antimicrobial peptide AP2 using SUMO fusion partner technology inEscherichia coli

Antimicrobial Peptides: Recent Insights on Biotechnological Interventions and Future Perspectives

A Toxicity Screening Approach to Identify Bacteriophage-Encoded Anti-Microbial Proteins

Contact Info

Product

Resources

About