Identification and characterization of a novel antimicrobial protein from the housefly Musca domestica

Guo, Guo; Ruyu, Tao; Li, Yan; Ma, Hui; Xiu, Jiangfan; Fu, Ping; Wu, Jianwei

doi:10.1016/j.bbrc.2017.06.112

Cited by 25 publications

(24 citation statements)

References 21 publications

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“…Antifungal activity of AMP-17. In a previous study, AMP-17 was shown to have strong antifungal activity against C. albicans ATCC76615, with a MIC of 2 μg/ml (Guo et al 2017). To further validate its antifungal effect against several common clinical pathogens, actively growing fungal cells of C. albicans, C. tropicalis, C. krusei, C. parapsilosis, and C. neoformans) were treated with purified AMP-17 protein.…”

Section: Resultsmentioning

confidence: 99%

“…AMP-17 protein was obtained from the Guoguo research team. It was produced in a prokaryotic expression system and purified by a nickel ion metal chelator affinity chromatography (Guo et al 2017). Fluco nazole (FLC), Sabouraud dextrose agar (SDA) and Sabouraud dextrose broth (SDB) were purchased from Solarbio (Beijing, China).…”

Section: Experimental Materials and Methodsmentioning

confidence: 99%

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Antifungal Activity and Physicochemical Properties of a Novel Antimicrobial Protein AMP-17 from Musca domestica

Yang

Guo

Zhao

et al. 2019

Polish Journal of Microbiology

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Antimicrobial peptides (AMPs) are cationic small peptide chains that have good antimicrobial activity against a variety of bacteria, fungi, and viruses. AMP-17 is a recombinant insect AMP obtained by a prokaryotic expression system. However, the full antifungal activity, physicochemical characteristics, and cytotoxicity of AMP-17 were previously unknown. AMP-17 was shown to have good antifungal activity against five pathogenic fungi, with minimum inhibitory concentrations (MIC) of 9.375–18.75 μg/ml, and minimum fungicidal concentrations (MFC) of 18.75–37.5 μg/ml. Notably, the antifungal activity of AMP-17 against Cryptococcus neoformans was superior to that of other Candida spp. In addition, the hemolytic rate of AMP-17 was only 1.47%, even at the high concentration of 16 × MIC. AMP-17 was insensitive to temperature and high salt ion concentration, with temperatures of 98°C and –80°C, and NaCl and MgCl2 concentrations of 50–200 mmol/l, having no significant effect on antifungal activity. However, AMP-17 was sensitive to proteases, trypsin, pepsin, and proteinase K. The elucidation of antifungal activity, physicochemical properties and cytotoxicity of AMP-17 provided an experimental basis for its safety evaluation and application, as well as indicated that AMP-17 might be a promising drug.

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Section: Resultsmentioning

confidence: 99%

Section: Experimental Materials and Methodsmentioning

confidence: 99%

Antifungal Activity and Physicochemical Properties of a Novel Antimicrobial Protein AMP-17 from Musca domestica

Yang

Guo

Zhao

et al. 2019

Polish Journal of Microbiology

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“…This is of particular importance in view of the increasing occurrence of multidrug resistant pathogens towards conventional antibiotics (Tonk and Vilcinskas, 2017). In the house fly and the black soldier fly the existence of anti-microbial peptides has already been shown (Elhag et al, 2017;Guo et al, 2017). Also, the effects on plant health of using the left-over substrates, containing chitin, as fertiliser need further investigation.…”

Section: Discussionmentioning

confidence: 99%

Insects as food and feed, a new emerging agricultural sector: a review

Huis

2020

JIFF

318

148

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During the last five years the scientific knowledge on insects as food and feed has been growing exponentially. At the same time, the industrial sector is increasingly engaged in rearing, processing and marketing of edible insects. Considerable attention is given to the black soldier fly as it can convert organic waste streams and transform it into several feed, food and industrial products. The farming of insects has an environmental impact which is lower than that of livestock species. The profitability of industrial production of insects as feed depends very much on the availability and applicability of cheap non-utilised side-streams. Microbial communities and their relationship with insects deserve full attention as it may help in the conversion of organic side streams of low economic value. Nutrition and health benefits for animals and humans need further exploration, also considering that insects have the largest anti-microbial peptide reservoir of all animals. Plant health can also be promoted by using chitin-containing leftover substrates as fertiliser. As insects have only recently been considered as food or feed, legislation trails developments. Therefore, politicians need to be assured that rearing and processing techniques are such that insect products are guaranteed free of chemical and microbial contaminants. Consumers are becoming more and more aware that insects as food are a viable option. Insects need to be processed into ingredients, that can be applied for safe and appetising products. The insect sector is maturing fast, but still faces many challenges, which can only be met when all stakeholders closely cooperate.

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“…The purified recombinant protein AMP-17 showed excellent antifungal activity in vitro. 14 – 16 However, the mechanism by which AMP-17 exerts antifungal effects is still unclear. To answer this question, we conducted an in-depth study on the potential anti- Candida mechanism of AMP-17 from the perspective of its influence on the cell wall integrity and cell membrane structure of C. albicans.…”

Section: Introductionmentioning

confidence: 99%

<p>Antimicrobial Peptide AMP-17 Affects <em>Candida albicans</em> by Disrupting Its Cell Wall and Cell Membrane Integrity</p>

Ma¹,

Zhao²,

Yang³

et al. 2020

IDR

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Background: Candida albicans is associated with high mortality among immunocompromised patients. Resistance to and toxic side effects of antifungal drugs require the development of alternative antifungal agents. AMP-17 is a novel antimicrobial peptide derived from Musca domestica that exerts excellent antifungal effects against the Candida species. In this article, we discuss the potential mechanism of AMP-17 against C. albicans from the perspective of affecting the latter's cell external structure. Methods: Recombinant AMP-17 was prepared by prokaryotic expression system, and its anti-C. albicans activity was detected by microdilution method. Microscopy and scanning electron microscopy were used to examine morphological changes in C. albicans. Cell wallspecific staining method was used to detect the change of cell wall integrity of C. albicans after AMP-17 treatment. AMP-17-induced damage to the C. albicans cell membrane was analyzed by fluorescent probes and glycerol assay kit. The expression of genes related to fungal cell wall and cell-membrane synthesis was detected by qRT-PCR. Results: Morphological observations showed that the growth of C. albicans was significantly inhibited in AMP-17-treated cells; the cells appeared aggregated and dissolved, with severe irregularities in shape. Furthermore, AMP-17 damaged the integrity of C. albicans cell walls. The cell wall integrity rate of AMP-17-treated cells was only 21.7% compared to untreated cells. Moreover, the change of membrane dynamics and permeability suggested that the cell membrane was disrupted by AMP-17 treatment. Genetic analysis showed that after AMP-17 treatment, the cell wall synthesis-related gene FKS2 of C. albicans was upregulated 3.46-fold, while the cell membrane ergosterol synthesis-related genes ERG1, ERG5, ERG6, and MET6 were down-regulated 5.88-, 17.54-, 13.33-, and 7.14-fold, respectively. Conclusion: AMP-17 treatment disrupted the cell wall integrity and membrane structure of C. albicans and is likely a novel therapeutic option for prevention and control of C. albicans infections.

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Identification and characterization of a novel antimicrobial protein from the housefly Musca domestica

Cited by 25 publications

References 21 publications

Antifungal Activity and Physicochemical Properties of a Novel Antimicrobial Protein AMP-17 from Musca domestica

Antifungal Activity and Physicochemical Properties of a Novel Antimicrobial Protein AMP-17 from Musca domestica

Insects as food and feed, a new emerging agricultural sector: a review

<p>Antimicrobial Peptide AMP-17 Affects <em>Candida albicans</em> by Disrupting Its Cell Wall and Cell Membrane Integrity</p>

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