Antimicrobial activity of biocomposite films containing cellulose nanofibrils and ethyl lauroyl arginate


Food packaging is tailored to keep food fresh by increasing shelf life and preventing microbial deterioration. However, traditional food packaging is commonly made from non-degradable polymers without antimicrobial properties and that pose an environmental threat if not disposed properly. To address this issue, here we describe the preparation of cellulose nanofibril (CNF) films and hydrogels with antimicrobial activity against common foodborne pathogens such as verotoxigenic E. coli, L. monocytogenes and S. Typhimurium. Furthermore, two grades of negatively charged CNFs with different fibrillation degrees were modified with ethyl lauroyl arginate (LAE), which is an antimicrobial agent. CNF films were able to bind LAE molecules up to a maximum concentration of 145–160 ppm. LAE–CNF biocomposite films exerted a bactericidal activity against a major foodborne pathogen present in ready-to-eat food (L. monocytogenes) even at 1% LAE. Our work describes a novel biopolymer-based strategy that overcomes the current hurdles with LAE incorporation into packaging materials, offering a green and antimicrobial alternative for packaging of ready-to-eat (RTE) meat products.

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  1. 1

    Khan A, Huq T, Khan RA et al (2014) Nanocellulose-based composites and bioactive agents for food packaging. Crit Rev Food Sci Nutr 54:163–174

    Article  CAS  Google Scholar 

  2. 2

    Liimatainen H, Visanko M, Sirviö JA et al (2012) Enhancement of the nanofibrillation of wood cellulose through sequential periodate–chlorite oxidation. Biomacromol 13:1592–1597

    Article  CAS  Google Scholar 

  3. 3

    Wagberg L, Decher G, Norgren M et al (2008) The build-up of polyelectrolyte multilayers of microfibrillated cellulose and cationic polyelectrolytes. Langmuir 24:784–795

    Article  CAS  Google Scholar 

  4. 4

    Saito T, Nishiyama Y, Putaux JL et al (2006) Homogeneous suspensions of individualized microfibrils from TEMPO-catalyzed oxidation of native cellulose. Biomacromol 7:1687–1691

    Article  CAS  Google Scholar 

  5. 5

    De France KJ, Hoare T, Cranston ED (2017) Review of hydrogels and aerogels containing nanocellulose. Chem Mater 29:4609–4631

    Article  CAS  Google Scholar 

  6. 6

    Chinga-Carrasco G, Syverud K (2012) On the structure and oxygen transmission rate of biodegradable cellulose nanobarriers. Nanoscale Res Lett 7:192–198

    Article  CAS  Google Scholar 

  7. 7

    Fukuzumi H, Saito T, Wata T et al (2009) Transparent and high gas barrier films of cellulose nanofibers prepared by TEMPO-mediated oxidation. Biomacromol 10:162–165

    Article  CAS  Google Scholar 

  8. 8

    Powell LC, Khan S, Chinga-Carrasco G et al (2016) An investigation of Pseudomonas aeruginosa biofilm growth on novel nanocellulose fibre dressings. Carbohydr Polym 137:191–197

    Article  CAS  Google Scholar 

  9. 9

    Jack AA, Nordli HR, Powell LC et al (2017) The interaction of wood nanocellulose dressings and the wound pathogen P. aeruginosa. Carbohydr Polym 157:1955–1962

    Article  CAS  Google Scholar 

  10. 10

    de Castro DO, Tabary N, Martel B et al (2018) Controlled release of carvacrol and curcumin: bio-based food packaging by synergism action of TEMPO-oxidized cellulose nanocrystals and cyclodextrin. Cellulose 25:1249–1263

    Article  CAS  Google Scholar 

  11. 11

    Weishaupt R, Heuberger L, Siqueira G et al (2018) Enhanced antimicrobial activity and structural transitions of a nanofibrillated cellulose-nisin biocomposite suspension. ACS Appl Mater Interfaces 10:20170–20181

    Article  CAS  Google Scholar 

  12. 12

    Nerin C, Becerril R, Manso S, Silva F (2016) Ethyl Lauroyl arginate (LAE): antimicrobial activity and applications in food systems. A2-Barros-Velázquez, Jorge. In: Antimicrobial food packaging. Academic Press, San Diego, pp 305–312

  13. 13

    Otero V, Becerril R, Santos JA et al (2014) Evaluation of two antimicrobial packaging films against Escherichia coli O157:H7 strains in vitro and during storage of a Spanish ripened sheep cheese (Zamorano). Food Control 42:296–302

    Article  CAS  Google Scholar 

  14. 14

    Theinsathid P, Visessanguan W, Kruenate J et al (2012) Antimicrobial activity of lauric arginate-coated polylactic acid films against Listeria monocytogenes and Salmonella Typhimurium on cooked sliced ham. J Food Sci 77:M142–M149

    Article  CAS  Google Scholar 

  15. 15

    Moreno O, Atarés L, Chiralt A et al (2018) Starch-gelatin antimicrobial packaging materials to extend the shelf life of chicken breast fillets. LWT-Food Sci Technol 97:483–490

    Article  CAS  Google Scholar 

  16. 16

    Kashiri M, Cerisuelo JP, Domínguez I et al (2016) Novel antimicrobial zein film for controlled release of lauroyl arginate (LAE). Food Hydrocoll 61:547–554

    Article  CAS  Google Scholar 

  17. 17

    Haghighi H, De Leo R, Bedin E et al (2019) Comparative analysis of blend and bilayer films based on chitosan and gelatin enriched with LAE (lauroyl arginate ethyl) with antimicrobial activity for food packaging applications. Food Packag Shelf Life 19:31–39

    Article  Google Scholar 

  18. 18

    Muriel-Galet V, Carballo GL, Hernández-Muñoz P, Gavara R (2016) Chapter 24-Ethyl lauroyl arginate (LAE): usage and potential in antimicrobial packaging A2-barros-Velázquez, Jorge. Antimicrobial food packaging. Academic Press, San Diego, pp 313–318

    Google Scholar 

  19. 19

    Chinga-Carrasco G, Kuznetsova N, Garaeva M et al (2012) Bleached and unbleached MFC nanobarriers: properties and hydrophobisation with hexamethyldisilazane. J Nanoparticle Res 14:1280–1284

    Article  CAS  Google Scholar 

  20. 20

    Saito T, Isogai A (2004) TEMPO-mediated oxidation of native cellulose. The effect of oxidation conditions on chemical and crystal structures of the water-insoluble fractions. Biomacromol 5:1983–1989

    Article  CAS  Google Scholar 

  21. 21

    Inoue T, Shingaki R, Hirose S et al (2007) Genome-wide screening of genes required for swarming motility in Escherichia coli K-12. J Bacteriol 189:950–957

    Article  CAS  Google Scholar 

  22. 22

    Di Bonaventura G, Piccolomini R, Paludi D et al (2008) Influence of temperature on biofilm formation by Listeria monocytogenes on various food-contact surfaces: relationship with motility and cell surface hydrophobicity. J Appl Microbiol 104:1552–1561

    Article  Google Scholar 

  23. 23

    Reeser RJ, Medler RT, Billington SJ et al (2007) Characterization of Campylobacter jejuni biofilms under defined growth conditions. Appl Environ Microbiol 73:1908–1913

    Article  CAS  Google Scholar 

  24. 24

    Chen CY, Nace GW, Irwin PL (2003) A 6 × 6 drop plate method for simultaneous colony counting and MPN enumeration of Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli. J Microbiol Methods 55:475–479

    Article  CAS  Google Scholar 

  25. 25

    Pezo D, Navascues B, Salafranca J, Nerin C (2012) Analytical procedure for the determination of Ethyl Lauroyl Arginate (LAE) to assess the kinetics and specific migration from a new antimicrobial active food packaging. Anal Chim Acta 745:92–98

    Article  CAS  Google Scholar 

  26. 26

    Chinga-Carrasco G, Averianova N, Kondalenko O et al (2014) The effect of residual fibres on the micro-topography of cellulose nanopaper. Micron 56:80–84

    Article  CAS  Google Scholar 

  27. 27

    Tehrani Z, Nordli HR, Pukstad B et al (2016) Translucent and ductile nanocellulose-PEG bionanocomposites—a novel substrate with potential to be functionalized by printing for wound dressing applications. Ind Crops Prod 93:193–202

    Article  CAS  Google Scholar 

  28. 28

    Mikczinski MR, Josefsson G, Chinga-Carrasco G et al (2014) Nanorobotic testing to assess the stiffness properties of nanopaper. IEEE Trans Robot 30:115–119

    Article  Google Scholar 

  29. 29

    Josefsson G, Chinga-Carrasco G, Gamstedt EK (2015) Elastic models coupling the cellulose nanofibril to the macroscopic film level. RSC Adv 5:58091–58099

    Article  CAS  Google Scholar 

  30. 30

    Sun FZ, Nordli HR, Pukstad B et al (2017) Mechanical characteristics of nanocellulose-PEG bionanocomposite wound dressings in wet conditions. J Mech Behav Biomed Mater 69:377–384

    Article  CAS  Google Scholar 

  31. 31

    Weller-Stuart T, Toth I, De Maayer P, Coutinho T (2017) Swimming and twitching motility are essential for attachment and virulence of Pantoea ananatis in onion seedlings. Mol Plant Pathol 18:734–745

    Article  CAS  Google Scholar 

  32. 32

    Josenhans C, Suerbaum S (2002) The role of motility as a virulence factor in bacteria. Int J Med Microbiol 291:605–614

    Article  CAS  Google Scholar 

  33. 33

    Morales-Soto N, Anyan ME, Mattingly AE et al (2015) Preparation, imaging, and quantification of bacterial surface motility assays. J Vis Exp 98:52338–52348

    Google Scholar 

  34. 34

    Lippolis JD, Brunelle BW, Reinhardt TA et al (2014) Proteomic analysis reveals protein expression differences in Escherichia coli strains associated with persistent versus transient mastitis. J Proteomics 108:373–381

    Article  CAS  Google Scholar 

  35. 35

    Allison SE, Silphaduang U, Mascarenhas M et al (2012) Novel repressor of Escherichia coli O157:H7 motility encoded in the putative fimbrial cluster OI-1. J Bacteriol 194:5343–5352

    Article  CAS  Google Scholar 

  36. 36

    O’Neil HS, Marquis H (2006) Listeria monocytogenes flagella are used for motility, not as adhesins, to increase host cell invasion. Infect Immun 74:6675–6681

    Article  CAS  Google Scholar 

  37. 37

    Barbosa FD, Neto OCD, Batista DFA et al (2017) Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken. Braz J Microbiol 48:754–759

    Article  CAS  Google Scholar 

  38. 38

    Kearns DB (2010) A field guide to bacterial swarming motility. Nat Rev Microbiol 8:634–644

    Article  CAS  Google Scholar 

  39. 39

    de la Fuente-Nunez C, Korolik V, Bains M et al (2012) Inhibition of bacterial biofilm formation and swarming motility by a small synthetic cationic peptide. Antimicrob Agents Chemother 56:2696–2704

    Article  CAS  Google Scholar 

  40. 40

    Guttenplan SB, Kearns DB (2013) Regulation of flagellar motility during biofilm formation. FEMS Microbiol Rev 37:849–871

    Article  CAS  Google Scholar 

  41. 41

    Belas R (2013) When the swimming gets tough, the tough form a biofilm. Mol Microbiol 90:1–5

    CAS  Google Scholar 

  42. 42

    Rees A, Powell LC, Chinga-Carrasco G et al (2015) 3D Bioprinting of carboxymethylated-periodate oxidized nanocellulose constructs for wound dressing applications. Biomed Res Int 2015:925757–925764

    Article  CAS  Google Scholar 

  43. 43

    Ma Q, Davidson PM, Zhong Q (2013) Antimicrobial properties of lauric arginate alone or in combination with essential oils in tryptic soy broth and 2% reduced fat milk. Int J Food Microbiol 166:77–84

    Article  CAS  Google Scholar 

  44. 44

    Becerril R, Manso S, Nerin C et al (2013) Antimicrobial activity of lauroyl arginate ethyl (LAE), against selected food-borne bacteria. Food Control 32:404–408

    Article  CAS  Google Scholar 

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The Research Council of Norway is acknowledged for the support to the Norwegian Micro- and Nano-Fabrication Facility, NorFab, project number 245963/F50. Mirjana Filipovic at RISE PFI is acknowledged for skilful assistance in the preparation of the CNF materials. During part of this work, Filomena Silva acknowledged a postdoctoral fellowship [grant number SFRH/BPD/79250/2011] from Fundação para a Ciência e Tecnologia within the scope of QREN—POPH—Advanced Formation programs co-funded by Fundo Social Europeu and MEC. This work was partially funded by FEDER funds through Programa Operacional Factores de Competitividade—COMPETE and by National Funds through FCT—Fundação para a Ciência e Tecnologia within the scope of project PEst-C/SAU/UI0709/2011. The authors also acknowledge the funding provided by the Ministerio de Economía, Industria y Competitividad (AGL-2015-67362-P), the Gobierno de Aragón (Spain) and Fondo Social Europeo to the GUIA group [financiación grupo GUIA T-10]. The “COST” Action (European Cooperation in Science and Technology) FP1405 “Active and intelligent (fibre-based) packaging—innovation and market introduction” is acknowledged for providing the channel for this collaboration.

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Correspondence to Gary Chinga-Carrasco.

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Silva, F., Gracia, N., McDonagh, B.H. et al. Antimicrobial activity of biocomposite films containing cellulose nanofibrils and ethyl lauroyl arginate. J Mater Sci 54, 12159–12170 (2019).

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