Synthesis and characterization of poly (3-hydroxybutyrate) films containing mentha piperita oil and antimicrobial activity assay by diffusion method


Food decomposition, spoilage and corruption caused by food borne microorganisms and non-biodegradable plastic materials for food packaging are the most important problems that threaten the environment and the economy. As a result, aim this study producing packaging materials with durable antibacterial properties against food spoilage microorganisms. To keep food fresh and a healthy environment. The effectiveness of this new active packaging material against four spoilage bacteria isolated from garden soil was tested by Mueller Hinton agar -diffusion method. A circles were cut from the film with approximately 0.6 cm in diameter and tested. The FTIR spectra of Mentha piperita oil with those of Poly (3-hydroxybutyrate) (P(3HB)) and it was possible to observe the characteristic bands and through which can be checked incorporation of the Mentha piperita oil into the polymer matrix can be verified. Microbiological analyzes showed that (P(3HB)) film with 10 % of oil better antimicrobial activity to Staphylococcus aureus with the inhibition zone 17.4 mm. Salmonella spp was more sensitive compared to Escherichia coli and Bacillus cereus with inhibition zones diameters 12.8, 8.5 and 5.6 mm, respectively. Through the previous results obtained in this study is a first step in the manufacture of cans and bags have the ability to biodegradation and antibacterial properties of microorganisms to obtain an environment add and free of waste.


  • Aggarwal KK, Khanuja SPS, Ahmad A, Kumar TRS, Gupta VK, Kumar S (2002) Antimicrobial activity profiles of the two enantiomers of limonene and carvone isolated from the oils of Mentha spicata and Anethum sowa. Flavour Frag J. 17(1): 59-63.
  • Antolak H, Czyzowska Kr ˛egie D . (2018). Activity of Mentha piperita L. Ethanol Extract against Acetic Acid Bacteria Asaia spp. Foods. 7: 171.
  • Aswathy M (2015) Production of Bio-Plastics (Polyhydroxy Butyrate) from Industrial Effluent using Batch and Two Stage Batch Culture Studies.Indian Journal of Science and Technology, 8(32).
  • Avila-Sosa R, Palou E, Munguía MTJ, Nevárez Moorillón GV, Navarro CAR, Malo AL (2012) Antifungal activity by vapor contact of essential oils added to amaranth, chitosan, or starch edible films. Int. J. Food Microbiol, 153: 66.
  • Bharti SN, Swetha G (2016) Need for Bioplastics and Role of Biopolymer PHB: A Short Review. J Pet Environ Biotechnol. 7: 272.
  • Bugnicourt E, Cinelli P, Lazzeri A, Alvarez V (2014) Polyhydroxyalkanoate (PHA): Review of synthesis, characteristics, processing and potential applications in packaging. Express Polym. Lett. 8: 791.
  • Derwich E, Chabir R, Taouil R, Senhaji O (2011) In vitro antioxidant activity and GC/MS studies on the leaves of Mentha piperita (Lamiaceae) from Morocco. IJPSDR.3(2): 130-6.
  • Giaquinto CDM,de SouzaGKM, Caetano VF, Vinhas GM (2017) Evaluation of the mechanical and thermal properties of PHB/canola oil films. Polímeros. 27(3): 201-207.
  • Heitmann AP, Patrício PSO, Coura IR, Pedroso EF, Souza PP, Mansur HS, Mansur A, Oliveira LCA (2016) Nanostructured niobium oxyhydroxide dispersed Poly(3-hydroxybutyrate) (PHB) films: Highly efficient photocatalystsfordegradation methylene blue dye. Applied Catalysis B: Environmental, 189: 141-150.
  • Huang T, Qian Y, Wei J, Zhou C (2019) Polymeric Antimicrobial Food Packaging and Its Applications. Polymers.11: 560.
  • Kadhim NH, Oda NA (2014) Determination of optimum condition for Poly-β-Hydroxybutyrate Production from Bacillus spp. Kerbala Journal of Pharmaceutical Sciens. 7.
  • Kiran GS, Jackson SA, Priyadharsini S, Dobson ADW, Selvin J (2017) Synthesis of Nm-PHB (nanomelanin-polyhydroxy butyrate) nanocomposite flm and its protective effect against bioflm forming multi drug resistant Staphylococcus aureus. Scientific REPOrtS. 7: 9167.
  • Kumar D, Kumar P, Pandey J (2018) Binary grafted chitosan film: Synthesis, characterization, antibacterial activity and prospects for food packaging. Int. J. Biol. Macromol. 115: 341-348.
  • Logan NA, Vose PD (2009) Bergey’s Manual Systematic Bacteriology Second Edition. Volume Three. Newyork: Springer P. 21-128.
  • Mahboubi M, Haghi G (2008) Antimicrobial activity and chemical composition of Mentha pulegium L. essential oil. J Ethnopharmacol.119(2): 325-7.
  • Majdizadeh M, Rezaei ZS, Alsadat MA, Shahi MH, Sasani E, Haghiralsadat BF (2018) A new strategy in improving therapeutic indexes of medicinal herbs: preparation and characterization of nanoliposomes containing Mentha piperita essential oil. J Shahid Sadoughi Univ Med Sci. 25(11): 853-64.
  • Morelli CL, Mahrous M, Belgacem MN, Branciforti MC, Suman Bretas RE, Bras J (2015) Natural copaiba oil as antibacterial agent for bio-based active packaging. Ind. Crop. Prod. 70: 134-141.
  • NCCLS (2012) Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard - Eleventh Edition.
  • Okmen AS, Okmen G, Arslan A, Vurkun M (2017) Antibacterial Activities of Mentha piperita L. Extracts Against Bacteria Isolated from Soccer Player’s Shoes and its Antioxidant Activities. Indian Journal of Pharmaceutical Education and Research: 51(3).
  • Park Y, Jin S, Park Y, Kim SM, NodaI, Chae B, Jung YM (2019) Studies on Chemical IR Images of Poly(hydroxybutyrate-co-hydroxyhexanoate)/ Poly(ethylene glycol) Blends and Two-Dimensional Correlation Spectroscopy. Polymers,11: 507.
  • Recha CR, Martellib SM, Silvaa BEB, Brabes KCS (2018) Antimicrobial Analysis and Characterization of P(3HB) Films Containing Essential Oils. Orbital: The Electronic Journal of Chemistry, 10(1).
  • Sabarinathan D, Chandrika SP, Venkatraman P, Easwaran M, Sureka CS, Preethia K. (2018). Production of polyhydroxybutyrate (PHB) from Pseudomonas plecoglossicida and its application towards cancer detection. Informatics in Medicine Unlocked, 11: 61-67.
  • Sadowska U, Matwijczuk A, Niemczynowicz A, Dróżdż T, Żabiński A (2019) Spectroscopic Examination and Chemometric Analysis of Essential Oils Obtained from Peppermint Herb (Mentha piperita L.) and Caraway Fruit (Carum carvi L.) Subjected to Pulsed Electric Fields.Processes 7(7): 466;
  • Singh CS, Agarwal R (2013) Evaluation of antibacterial activity of volatile oil from Mentha spicata L. JDDT. 3(4): 120-1.
  • Singh R, Shushni MAM, Belkheir A (2015) Antibacterial and antioxidant activities of Mentha piperita L. Arabian Journal of Chemistry 8: 322-328.
  • Sobouti B, Ghavami Y, Asadifar B, Jafarzadeh M Ghelman M, Vaghardoost R (2020) Determination of Serum Levels of Interleukin-6, Interleukin-8, Interleukin-10, and Tumor Necrosis-Alpha and their Relationship With The Total Body Surface Area in Children. Journal of Burn Care & Research. 41(3): 539-543
  • Soković MD, Vukojević J, Marin PD, Brkić DD, Vajs V, Van Griensven LJ (2009) Chemical composition of essential oils of Thymus and Mentha species and their antifungal activities. Molecules. 14(1): 238-49.
  • Sujana P, Sridhar TM, Josthna P, Naidu CV (2013) Antibacterial Activity and Phytochemical Analysis of Mentha piperita L. (Peppermint)—An Important Multipurpose Medicinal Plant. American Journal of Plant Sciences. 4: 77-83.
  • Talebi F, Misaghi A, Khanjari A, Kamkar A, Gandomi H, Rezaeigolestani M (2018) Incorporation of spice essential oils into poly-lactic acid film matrix with the aim of extending microbiological and sensorial shelf life of ground beef. LWT-Food. Sci. Technol. 96: 482-490.
  • Thapa C, Shakya P, Shrestha R, Pal S, Manandhar P (2018) Isolation of Polyhydroxybutyrate (PHB) Producing Bacteria, Optimization of Culture Conditions for PHB production, Extraction and Characterization of PHB. Nepal Journal of Biotechnology. 6(1): 62-68.
  • Tong SY, Lim PN, Wang K, Thian ES (2018) Development of a functional biodegradable composite with antibacterial properties. Mater. Technol. 33: 754-759.
  • Vasile C, Sivertsvik M, Mitelut AC, Brebu MA, Stoleru E, Rosnes JT, Tanase EE, Khan W, Pamfil D, Cornea CP (2017) Comparative Analysis of the Composition and Active Property Evaluation of Certain Essential Oils to Assess their Potential Applications in Active Food Packaging. Materials 10: 45.
  • Vergis J, Gokulakrishnan P, Agarwa RK, Kumar A (2015) Essential Oils as Natural Food Antimicrobial Agents: A Review. Crit. Rev. Food Sci. Nutr. 55: 1320-1323.
  • Wellen RMR, Rabello MS, Fechine GJM, Canedo EL (2013) The melting behaviour of poly(3-hydroxybutyrate) by DSC. Reproducibility study. Polym. Test.32: 215.
  • Yu Z, Li B, Chu J, Zhang P (2018) Silica in situ enhanced PVA/chitosan biodegradable films for food packages. Carbohydr. Polym.184: 214-220.


This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.