Characterization and antibacterial activity of the nanocomposite of half-fin anchovy (Setipinna taty) hydrolysates/zinc oxide nanoparticles
Graphical abstract
Introduction
Zinc oxide (ZnO) is a kind of wide band gap (3.37 eV) semiconductor with large exciton binding energy (60 meV) at room temperature [1]. ZnO is generally regarded as a safe (GRAS) material by the United States Food and Drug Administration (USFDA, 21CFR182.8991) [2]. The worldwide production of ZnO is estimated at 0.1–1.2 million tons per year [3], [4]. Due to physical and chemical stabilities, high biocompatibility, lower cost, and nontoxicity, ZnO and their nanoparticles (ZnO NPs) have unique advantages over other nano metal oxides, and thereby are extensively used in daily life products such as cosmetics, medical devices, electronic sensor, lubricants, etc. [5].
Nano structured materials have been used in food industry to inhibit the growth of bacteria present on the surface of food products [6]. The most commonly used polymers are polysaccharides and proteins in biocomposite [7]. The properties of polymer, such as thermal, mechanical, and gas barrier, can be enhanced by addition of nanoparticles [8], [9], [10], [11]. The antimicrobial property of ZnO NPs would make them a viable approach to prevent infectious diseases [12]. The large specific surface area and high surface energy of ZnO NPs will contribute to their excellent interfacial interactions on polymer branches [13]. To date, ZnO NPs have been explored as functional nanofiller incorporated into packaging and food contact materials to inhibit the target bacteria [14], [15], [16]. For example, the polymer incorporation of ZnO NPs as active packaging has been reported to extend the shelf life or to enhance quality for fresh orange juice [17], fresh-cut ‘Fuji’ apple [18], convenient vegetable dishes [19], postharvest strawberry [20], soft white cheese [21], [22], and ready-to-eat poultry meat [23].
Recently, researchers have been focusing on the conjugates of nanoparticles with antimicrobial peptides (AMPs) as a promising approach to achieve superior antimicrobial activity. Mei et al. found that the silver nanoparticles (AgNPs) functionalized with both bacitracin A and polymyxin E (AgNPs-BA&PE) easily attached and penetrated into the bacterial cell membrane, resulting in up to 10-fold increase in the antibacterial activity without the emergence of bacterial resistance [24]. In a latest study by Pal et al., who confirmed the activity and stability of cysteine containing AMPs were enhanced after conjugation with AgNPs [25]. The conjugate of gold nanoparticles (AuNPs) and peptide PEP (H-Cys-Ala-Cys-Trp-Gln-Val-Ser-Arg-Arg-Arg-Arg-Gly-OH) integrated the advantages of cationic AuNPs and AMPs as unique non-viral vectors for gene delivery to mesenchymal stem cells [26]. The functionalized ZnO NPs with glucose oxidase for specific biosensors [27], and with β-galactosidase or α-amylase for enhanced heat-resistance and stability [28], [29] have been reported compared to the corresponding free enzymes. However, to the best of our knowledge, there are few studies about antibacterial protein hydrolysates or peptides incorporated with ZnO NPs.
Half-fin anchovy (Setipinna taty) hydrolysates (HAHp) derived from the digestion of half-fin anchovy are composed of peptides and amino acids [30]. HAHp demonstrated antibacterial, antioxidant, and antiproliferative activities [31], [32]. In this study, HAHp were conjugated with ZnO NPs via a simple and green hydrothermal method to fabricate a novel nanocomposite. The property of the nanocomposite was evaluated. Furthermore, the antibacterial effects of the nanocomposite were determined using Escherichia coli as the indicative strain. All the results will provide references for the adsorption of peptide and/or amino acids onto ZnO NPs, and further deplore a novel antibacterial agent derived from HAHp used in foods.
Section snippets
Materials
Half-fin anchovy (Setipinna taty), were obtained from Fengmao aquatic market in Zhoushan City, China. The bacteria strains of Escherichia coli (E. coli) (CGMCC 1.1100), Pseudomonas fluorescens (P. fluorescens) (CICC 20225), Staphylococcus aureus (S. aureus) (CMCC 26003), Salmonella (CICC 10982), and Bacillus cereus (B. cereus) (CICC 10809) were storage strains in College of Food and Pharmacy, Zhejiang Ocean University. Erythromycin was purchased from Aladdin in China. O-Nitro-phenyl-β-d
Micromorphology and total Zinc content of HAHp(3.0)/ZnO NPs
The ZnO crystals used in this study were synthesized by a low temperature wet chemical method. The TEM image of Fig. 1A revealed that spherical and rhomboid shapes were observed in ZnO crystals. The particle sizes were 10–40 nm. The pattern of these crystals displayed well-defined diffraction peaks at 2θ of 31.83°, 34.49°, 36.32°, 47.59°, 56.52°, 62.87°, 66.39°, 68.00°, 69.10°, 72.56°, and 76.95°, as observed in Fig. 1B, which are indicated as (100), (002), (101), (102), (110), (103), (200),
Conclusions
The nanocomposite of HAHp(3.0)/ZnO NPs was obtained by a simple, low cost and green method. Results showed that the HAHp(3.0)/ZnO NPS demonstrated stronger inhibition effects on E. coli, P. fluorescens, Salmonella and S. aureus compared with the bare HAHp(3.0), according to their MIC values. Mechanistic investigation for the antibacterial effect revealed that the HAHp(3.0)/ZnO NPs caused disorganization in the bacterial cytomembrane and led to leakage of cytoplasmic contents, hence exhibited
Acknowledgments
This work was supported by grants from the Zhejiang Natural Science Project (LY15C200018), the Public Welfare Project of Zhejiang Province (2016C32066), the Zhoushan Science and Technology Bureau Project (2014C41005), and the National Natural Science Foundation of China (31301568). We also thank Prof. Zuisu Yang for assisting the operation of fluorescence microscopy.
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