Antibacterial properties of Acinetobacter baumanniiphage Abp1 endolysin (PlyAB1)

Huang, Guangtao; Shen, Xiaodong; Gong, Yanhong; Dong, Zhi-Wei; Zhao, Xia; Shen, Wei; Wang, Jing; Hu, Fuquan; Peng, Yizhi

doi:10.1186/s12879-014-0681-2

Cited by 50 publications

(44 citation statements)

References 27 publications

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“…In vitro experiments demonstrated that endolysins exert bactericidal activity against Gram-negative strains in concentrations varying from 1 to 500 µg/mL (0.06-6.2 µM) [18,20,23,34] and values significantly depended on the strains used. For LysECD7-8his, it was shown that less than 100 µg/mL (6.2 µM) was enough to eradicate growing bacteria up to five orders of magnitude.…”

Section: Discussionmentioning

confidence: 99%

“…This means that histidine tagging definitely affects the bactericidal properties of endolysin, but the charge hypothesis does not completely explain these alterations, and the particular effects of different lengths of fusions are difficult to predict. Most of the endolysins described today are 6-His fusions [2,6,[16][17][18][19][20][21][22]. How the activity of all these endolysins are modified by 6-His tag fusion has not yet been described in detail.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, researchers rarely try several tag modifications when purifying the target protein and look for alternative purification methods only if no (or few) biological effects are observed [14,15]. Most of the recombinant endolysins under investigation were purified and tested for antibacterial activity with fused histidine tags [2,6,[16][17][18][19][20][21][22][23]. However, how the activity of these endolysins is modified by polyhistidine tag fusion has not been specifically studied.…”

Section: Introductionmentioning

confidence: 99%

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Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion

et al. 2020

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The use of recombinant endolysins is a promising approach for antimicrobial therapy capable of counteracting the spread of antibiotic-resistant strains. To obtain the necessary biotechnological product, diverse peptide tags are often fused to the endolysin sequence to simplify enzyme purification, improve its ability to permeabilize the bacterial outer membrane, etc. We compared the effects of two different types of protein modifications on endolysin LysECD7 bactericidal activity in vitro and demonstrated that it is significantly modulated by specific permeabilizing antimicrobial peptides, as well as by widely used histidine tags. Thus, the tags selected for the study of endolysins and during the development of biotechnological preparations should be used with the appropriate precautions to minimize false conclusions about endolysin properties. Further, modifications of LysECD7 allowed us to obtain a lytic enzyme that was largely devoid of the disadvantages of the native protein and was active over the spectra of conditions, with high in vitro bactericidal activity not only against Gram-negative, but also against Gram-positive, bacteria. This opens up the possibility of developing effective antimicrobials based on N-terminus sheep myeloid peptide of 29 amino acids (SMAP)-modified LysECD7 that can be highly active not only during topical treatment but also for systemic applications in the bloodstream and tissues.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion

et al. 2020

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“…A. baumannii species genotypic identification was performed by PCR detection of the OXA-51 gene (Turton et al, 2006; Zander et al, 2013). Isolates lacking the OXA-51 gene were investigated further by sequence analysis of the 16S rRNA gene (Huang et al, 2013, 2014). …”

Section: Methodsmentioning

confidence: 99%

Multilocus Sequence Typing Analysis of Carbapenem-Resistant Acinetobacter baumannii in a Chinese Burns Institute

et al. 2016

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Acinetobacter baumannii is a leading pathogen responsible for nosocomial infections. The emergence of carbapenem-resistant A. baumannii (CRAB) has left few effective antibiotics for clinicians to use. To investigate the temporal evolutionary relationships among CRAB strains, we collected 248 CRAB isolates from a Chinese burns institute over 3 years. The prevalence of the OXA-23 gene was detected by polymerase chain reaction. Multilocus sequence typing was used to type the CRAB strains and eBURST was used to analyze their evolutionary relationships. Wound surfaces (41%), sputa (24%), catheters (15%), and bloods (14%) were the four dominant isolation sources. Except for minocycline (33.5%) and sulbactam/cefoperazone (74.6%), these CRAB strains showed high resistance rates (>90%) to 16 tested antibiotics. The 248 isolates fall into 26 sequence types (STs), including nine known STs and 17 unknown STs. The majority (230/248) of these isolates belong to clonal complex 92 (CC92), including eight isolates belonging to seven unreported STs. A new CC containing 11 isolates grouped into four new STs was identified. The OXA-23 gene was detected at high prevalence among the CRAB isolates and the prevalence rate among the various STs differed. The majority of the isolates displayed a close evolutionary relationship, suggesting that serious nosocomial spreading and nosocomial infections of CRAB have occurred in the burn unit. In conclusion, the main CC for CRAB in this Chinese burn unit remained unchanged during the 3-year study period, and a new CC was identified. CC92 was the dominant complex, and more attention should be directed toward monitoring the new CC we have identified herein.

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“…It has a genome size of 42 kb containing 51 genes, including an endolysin gene [14] as expected for a lytic phage. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Phage Abp1 Rescues Human Cells and Mice from Infection by Pan-Drug Resistant Acinetobacter Baumannii

Yin

Huang

Zhang

et al. 2017

Cell Physiol Biochem

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Background/Aims: As an “ESKAPE” pathogen, Acinetobacter baumannii is one of the leading causes of drug-resistant infections in humans. Phage therapy may be a useful strategy in treating infections caused by drug-resistant A. baumannii. Among 21 phage strains that were isolated and described earlier, we investigated the therapeutic efficacy of Abp1 because of its relatively wide host range. Methods: Phage stability assays were used to evaluate thermal and pH stability of Abp1. Abp1 was co-cultured with A. baumannii (AB1) over a range of multiplicities of infection to determine its bactericidal efficacy. HeLa or THP-1 cells were used in the cytotoxicity and protection assays. Finally, the therapeutic effects of Abp1 on local and systemic A. baumannii infection in mice were determined. Results: We found that Abp1 exhibits high thermal and pH stability and has a low frequency of lysogeny. Bacteriophage resistance also occurs at a very low frequency (3.51±0.46×10^-8), and Abp1 can lyse almost all host cells at a MOI as low as 0.1. Abp1 has no detectable cytotoxicity to HeLa or THP-1 cells as determined by LDH release assay. Abp1 can rescue HeLa cells from A. baumannii infection, even if introduced 2 hours post infection. In both local and systemic A. baumannii infection mouse models, Abp1 treatment exhibits good therapeutic effects. Conclusion: Abp1 is an excellent candidate for phage therapy against drug-resistant A. baumannii infections.

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Antibacterial properties of Acinetobacter baumanniiphage Abp1 endolysin (PlyAB1)

Cited by 50 publications

References 27 publications

Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion

Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion

Multilocus Sequence Typing Analysis of Carbapenem-Resistant Acinetobacter baumannii in a Chinese Burns Institute

Phage Abp1 Rescues Human Cells and Mice from Infection by Pan-Drug Resistant Acinetobacter Baumannii

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