Evidence map›Paper›PMID 30835077›Full record

ArticleProbiotics and antimicrobial proteins2019

Design and Unique Expression of a Novel Antibacterial Fusion Protein Cecropin B-Human Lysozyme to Be Toxic to Prokaryotic Host Cells.

Yeli Zhang, Yuhua Li, Liujiao Bian

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Article in Probiotics and antimicrobial proteins, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

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1citing papers in PubMed
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1 · What the graph read from it

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3 · Its place in the literature

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1 citing paper in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

3 authors.

Yeli ZhangCollege of Life Science, Northwest University, Xi'an, 710069, Shaan'xi, PR China.
Yuhua LiCollege of Life Science, Northwest University, Xi'an, 710069, Shaan'xi, PR China.
Liujiao BianCollege of Life Science, Northwest University, Xi'an, 710069, Shaan'xi, PR China. bianliujiao@sohu.com.ORCID http://orcid.org/0000-0001-8993-2696

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

A novel antibacterial fusion protein, cecropin B-human lysozyme (CB-hLyso), was designed and expressed in a prokaryotic system. The full-length CB gene was first synthesized and fused to the 5' end of the hLyso gene. The recombinant CB-hLyso was then subcloned in plasmid pET32a, and pET32a-CB-hLyso was transferred into Escherichia coli (E. coli) BL21(DE3) and BL21(DE3)pLysS. The results showed that in the original culture media, Luria-Bertani (LB) media and terrific broth (TB), at 37 or 25 °C, CB-hLyso was barely expressed; however, when the original culture medium was replaced with an equi-volume of fresh medium, obvious expression occurred in BL21(DE3)pLysS/pET32a-CB-hLyso at 25 °C, and the expression in TB (25%) was higher than that in LB (15%). Through a two-step chromatographic method consisting of Ni-chelated Sepharose Fast Flow affinity and Sephadex G-75 size-exclusion, the crude fusion CB-hLyso was isolated in a homogeneous form, and preliminary bacteriostasis experiments showed that the fusion CB-hLyso had a strong inhibitory effect on the growth of Staphylococci. This work provides useful insights into the design of novel fusion polypeptides with higher bacteriolytic activity and wider antimicrobial spectra and in the expression of polypeptide products that are toxic to prokaryotic host cells, eukaryotic host cells or insect cells. Graphical Abstract Schematic representation of expression vector pET-32a-CB-hLyso, with Factor Xa and Asn-Gly.

Indexed as

Anti-Bacterial AgentsCecropinsEscherichia coliGene ExpressionHumansMuramidaseRecombinant Fusion ProteinsStaphylococcusAnti-Bacterial AgentsCecropinsMuramidaseRecombinant Fusion ProteinsAntibacterial fusion proteinCecropin BDesignExpressionHuman lysozyme

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