Hand Sanitizer Gel Formulation with Laccase Enzyme as an Antibacterial Against Staphylococcus aureus and Escherichia coli
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Submitted
28 June 2022
28 June 2022
Published
30 November 2022
30 November 2022
Abstract
Laccase enzymes have been used widely in industrial fields such as textile, pulp, paper, food, cosmetic, and pharmaceutical industries. Laccase is used in toothpaste, mouthwash, deodorants, and soaps in personal care products. Previously, laccase enzymes had never been used for formulating hand sanitizer gel. This study aimed to determine the effect of the laccase enzyme on the physicochemical properties and the antibacterial potential of the hand sanitizer gel against pathogenic bacteria. Laccase enzyme was produced through fermentation using the fungus Trametes hirsuta EDN 082 with an activity of 0.032 U/mL. Hand sanitizer gel was made with the addition of laccase enzyme with varying concentrations of 4, 7, and 10% (v/v). The physicochemical test included organoleptic tests, pH evaluation, gel spreadability, and viscosity. The antibacterial was tested by the palm swab method. The gel physicochemical characteristics showed that the more laccase enzyme added, the more yellow the color produced, the less thick the shape, the wider the gel spreadability, and the lower the viscosity. The obtained pH ranged from 7.4 to 7.6. The best formulation of the hand sanitizer gel was achieved with the addition of a 7% (v/v) laccase enzyme. This formulation can reduce the number of bacteria colonies of Staphylococcus aureus and Escherichia coli on the palms with effectiveness above 95%. The laccase enzyme can be used as an active ingredient and antibacterial agent in the formulation of hand sanitizers.
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References
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4. Manaye G, Muleta D, Henok A, Asres A, Mamo Y, Feyissa D, Ejeta F, et al. Evaluation of the Efficacy of Alcohol-Based Hand Sanitizers Sold in Southwest Ethiopia. Infect Drug Resist. 2021;14:547-54. doi:10.2147/idr.s288852
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6. Lachenmeier DW. Safety evaluation of topical applications of ethanol on the skin and inside the oral cavity. J Occup Med Toxicol. 2008;3:26. doi:10.1186/1745-6673-3-26
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10. Rahmasari D, Hendradi E, Chasanah U. Formulation and evaluation of hand sanitizer gel containing infused binahong leaf (Anredera cordifolia) as antibacterial preparation. Farmasains J Farmasi Ilmu Kesehatan. 2020;5(1):23–30. doi:10.22219/farmasains.v5i1.13008
11. Maulana MR, Ariningrum ND, Nurjanah BAD, Harismah K. Uji Stabilitas Fisik Hand Sanitizer Antiseptik Berbasis Daun Stevia Dan Kulit Nanas (Ananas comosus (L.) Merr.). Prosiding SNPBS Seminar Nasional Pendidikan Biol Saintek. 2020;5:391–7.
12. Ariningrum ND, Anisa B, Nurjanah D, Maulana MR, Harismah K. Uji efektivitas gel hand sanitizer sebagai antiseptik tangan berbasis ekstrak daun trembesi (Albizia saman (Jacq.) Merr) dan stevia. Prosiding SNPBS Seminar Nasional Pendidikan Biol Saintek. 2020;5:506–13.
13. Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, et al. Applications of Microbial Enzymes in Food Industry. Food Technol Biotechnol. 2018;56(1):16-30. doi:10.17113/ftb.56.01.18.5491
14. Zerva A, Simic S, Topakas S, Nikodinovic-Runic J. Applications of microbial laccases: Patent review of the past decade (2009-2019). Catalysts. 2019;9(12):1023. doi:10.3390/catal9121023
15. Dana M, Khaniki GB, Mokhtarieh AA, Davarpanah SJ. Biotechnological and industrial applications of laccase: A review. J Appl Biotechnol Reports. 2017;4(4):675–9.
16. Janusz G, Pawlik A, Świderska-Burek U, Polak J, Sulej J, Jarosz-Wilkołazka A, et al. Laccase properties, physiological functions, and evolution. Int J Mol Sci. 2020;21(3):966. doi:10.3390/ijms21030966
17. Bertrand B, Martínez-Morales F, Trejo-Hernández MR. Fungal laccases: Induction and production. Rev Mex Ing Quim. 2013;12(3):473–88.
18. Barber-Zucker S, Mateljak I, Goldsmith M, Kupervaser M, Aldalde M, Fleishman SJ. Designed High-Redox Potential Laccases Exhibit High Functional Diversity. ACS Catal. 2022;12(21):13164-73. doi:10.1021/acscatal.2c03006
19. Osma JF, Toca-Herrera JL, Rodríguez-Couto S. Uses of laccases in the food industry. Enzyme Res. 2010;2010:918761. doi:10.4061/2010/918761
20. Ningsih F, Yanto DHY, Mangunwardoyo W, Anita SH, Watanabe T. Optimization of laccase production from a newly isolated Trametes sp. EDN134. IOP Conf Ser Earth Environ Sci. 2020;572:012024. doi:10.1088/1755-1315/572/1/012024
21. Anita SH, Ardianti FC, Oktaviani M, Sari FP, Nurhayat OD, Ramadhan KP, et al. Immobilization of laccase from Trametes hirsuta EDN 082 in light expanded clay aggregate for decolorization of Remazol Brilliant Blue R dye. Bioresour Technol Rep. 2020;12:100602. doi:10.1016/j.biteb.2020.100602
22. Yanto DHY, Guntoro MA, Nurhayat OD, Anita SH, Oktaviani M, Ramadhan KP, et al. Biodegradation and biodetoxification of batik dye wastewater by laccase from Trametes hirsuta EDN 082 immobilised on light expanded clay aggregate. 3 Biotech. 2021;11(5):247. doi:10.1007/s13205-021-02806-8
23. Noviardi H, Himawan HC, Anggraeni R. Formulasi dan aktivitas antibakteri sediaan gel hand sanitizer dari ekstrak etanol biji mangga harum manis (Mangifera indica L.) terhadap Escherichia coli dan Staphylococcus aureus. J Farmamedika (Pharmamedica J). 2018;3(1):1–9. doi:10.47219/ath.v3i1.20
24. Radji M, Suryadi H. Uji Efektivitas Antimikroba Beberapa Merek Dagang Pembersih Tangan Antiseptik. Pharm Sci Res. 2007;4(1):1–6.doi:10.7454/psr.v4i1.3408
25. Wolfe MK, Lantagne DS. A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens. J Vis Exp. 2017;124:55604. doi:10.3791/55604
26. Fallica F, Leonardi C, Toscano V, Santonocito D, Leonardi P, Puglia C. Assessment of Alcohol-Based Hand Sanitizers for Long-Term Use, Formulated with Addition of Natural Ingredients in Comparison to WHO Formulation 1. Pharmaceutics. 2021;13(4):571. doi:10.3390/pharmaceutics13040571
27. Dolorosa MT, Nurjanah, Purwaningsih S, Anwar E. Utilization of Kappaphycus alvarezii and Sargassum plagyophyllum from Banten as cosmetic creams. IOP Conf Ser Earth Environ Sci. 2020;404:012008. doi:10.1088/1755-1315/404/1/012008
28. Binder L, Mazál J, Petz R, Klang V, Valenta C. The role of viscosity on skin penetration from cellulose ether-based hydrogels. Skin Res Technol. 2019;25(5):725-34. doi:10.1111/srt.12709
29. Kulawik-Pióro A, Miastkowska M. Polymeric Gels and Their Application in the Treatment of Psoriasis Vulgaris: A Review. Int J Mol Sci. 2021;22(10):5124. doi:10.3390/ijms22105124
30. Booq RY, Alshehri AA, Almughem FA, Zaidan NM, Aburayan WS, Bakr AA, et al. Formulation and evaluation of alcohol-free hand sanitizer gels to prevent the spread of infections during pandemics. Int J Environ Res Public Health. 2021;18(12):6252. doi:10.3390/ijerph18126252
31. Febriani A, Syafriana V, Afriyando H, Djuhariah YS. The utilization of oil palm leaves (Elaeis guineensis Jacq.) waste as an antibacterial solid bar soap. IOP Conf Ser Earth Environ Sci. 2020;572:012038. doi:10.1088/1755-1315/572/1/012038
32. Malanovic N, Lohner K. Antimicrobial Peptides Targeting Gram-Positive Bacteria. Pharmaceuticals. 2016;9(3):59. doi:10.3390/ph9030059
33. Syafriana V, Febriani A, Suyatno S, Nurfitri N, Hamida F. Antimicrobial Activity of Ethanolic Extract of Sempur (Dillenia suffruticosa (Griff.) Martelli) Leaves against Pathogenic Microorganisms. Borneo J Pharm. 2021;4(2):135–44. doi:10.33084/bjop.v4i2.1870
34. Wijaya JI. Formulasi Sediaan Gel Hand Sanitizer dengan Bahan Aktif Triklosan 1,5% dan 2%. Calyptra J Ilmiah Mahasiswa Univ Surabaya. 2013;2(1):1–14.
35. Weatherly LM, Gosse JA. Triclosan exposure, transformation, and human health effects. J Toxicol Environ Health B Crit Rev. 2017;20(8):447-69. doi:10.1080/10937404.2017.1399306
36. Kryscio DR, Sathe PM, Lionberger R, Yu L, Bell MA, Jay M, et al. Spreadability measurements to assess structural equivalence (Q3) of topical formulations--a technical note. AAPS PharmSciTech. 2008;9(1):84-6. doi:10.1208/s12249-007-9009-5
37. Basiak E, Lenart A, Debeaufort F. How Glycerol and Water Contents Affect the Structural and Functional Properties of Starch-Based Edible Films. Polymers. 2018;10(4):412. doi:10.3390/polym10040412
Read More
2. Golin AP, Choi D, Ghahary A. Hand sanitizers: A review of ingredients, mechanisms of action, modes of delivery, and efficacy against coronaviruses. Am J Infect Control. 2020;48(9):1062-7. doi:10.1016/j.ajic.2020.06.182
3. Atolani O, Baker MT, Adeyemi OS, Olanrewaju IR, Hamid AA, Ameen OM, et al. COVID-19: Critical discussion on the applications and implications of chemicals in sanitizers and disinfectants. EXCLI J. 2020;19:785-99. doi:10.17179/excli2020-1386
4. Manaye G, Muleta D, Henok A, Asres A, Mamo Y, Feyissa D, Ejeta F, et al. Evaluation of the Efficacy of Alcohol-Based Hand Sanitizers Sold in Southwest Ethiopia. Infect Drug Resist. 2021;14:547-54. doi:10.2147/idr.s288852
5. Huffer S, Clark ME, Ning JC, Blanch HW, Clark DS. Role of alcohols in growth, lipid composition, and membrane fluidity of yeasts, bacteria, and archaea. Appl Environ Microbiol. 2011;77(18):6400-8. doi:10.1128/aem.00694-11
6. Lachenmeier DW. Safety evaluation of topical applications of ethanol on the skin and inside the oral cavity. J Occup Med Toxicol. 2008;3:26. doi:10.1186/1745-6673-3-26
7. Cahyaningtyas FD, Ukrima ZA, Nora N, Amaria A. Pemanfatan Ekstrak Biji Teratai Sebagai Bahan Aktif Antibakteri Untuk Pembuatan Hand Sanitizer. Indones Chem Appl J. 2019;3(1):7. doi:10.26740/icaj.v3n1.p7-13
8. Asngad A, Bagas AR, Nopitasari N. Kualitas Gel Pembersih Tangan (Handsanitizer) dari Ekstrak Batang Pisang dengan Penambahan Alkohol, Triklosan dan Gliserin yang Berbeda Dosisnya. Bioeksperimen J Penelitian Biol. 2018;4(2):61–70. doi:10.23917/bioeksperimen.v4i2.6888
9. Umarudin U, Surahmaida S, Syukrianto S, Wulansari SA, Nurhaliza S. Aplikasi Hand Sanitizer Kitosan Cangkang Bekicot Sebagai Antibakteri dan Upaya Preventif Covid 19. Simbiosa. 2020;9(2):107–17. doi:10.33373/sim-bio.v9i2.2669
10. Rahmasari D, Hendradi E, Chasanah U. Formulation and evaluation of hand sanitizer gel containing infused binahong leaf (Anredera cordifolia) as antibacterial preparation. Farmasains J Farmasi Ilmu Kesehatan. 2020;5(1):23–30. doi:10.22219/farmasains.v5i1.13008
11. Maulana MR, Ariningrum ND, Nurjanah BAD, Harismah K. Uji Stabilitas Fisik Hand Sanitizer Antiseptik Berbasis Daun Stevia Dan Kulit Nanas (Ananas comosus (L.) Merr.). Prosiding SNPBS Seminar Nasional Pendidikan Biol Saintek. 2020;5:391–7.
12. Ariningrum ND, Anisa B, Nurjanah D, Maulana MR, Harismah K. Uji efektivitas gel hand sanitizer sebagai antiseptik tangan berbasis ekstrak daun trembesi (Albizia saman (Jacq.) Merr) dan stevia. Prosiding SNPBS Seminar Nasional Pendidikan Biol Saintek. 2020;5:506–13.
13. Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, et al. Applications of Microbial Enzymes in Food Industry. Food Technol Biotechnol. 2018;56(1):16-30. doi:10.17113/ftb.56.01.18.5491
14. Zerva A, Simic S, Topakas S, Nikodinovic-Runic J. Applications of microbial laccases: Patent review of the past decade (2009-2019). Catalysts. 2019;9(12):1023. doi:10.3390/catal9121023
15. Dana M, Khaniki GB, Mokhtarieh AA, Davarpanah SJ. Biotechnological and industrial applications of laccase: A review. J Appl Biotechnol Reports. 2017;4(4):675–9.
16. Janusz G, Pawlik A, Świderska-Burek U, Polak J, Sulej J, Jarosz-Wilkołazka A, et al. Laccase properties, physiological functions, and evolution. Int J Mol Sci. 2020;21(3):966. doi:10.3390/ijms21030966
17. Bertrand B, Martínez-Morales F, Trejo-Hernández MR. Fungal laccases: Induction and production. Rev Mex Ing Quim. 2013;12(3):473–88.
18. Barber-Zucker S, Mateljak I, Goldsmith M, Kupervaser M, Aldalde M, Fleishman SJ. Designed High-Redox Potential Laccases Exhibit High Functional Diversity. ACS Catal. 2022;12(21):13164-73. doi:10.1021/acscatal.2c03006
19. Osma JF, Toca-Herrera JL, Rodríguez-Couto S. Uses of laccases in the food industry. Enzyme Res. 2010;2010:918761. doi:10.4061/2010/918761
20. Ningsih F, Yanto DHY, Mangunwardoyo W, Anita SH, Watanabe T. Optimization of laccase production from a newly isolated Trametes sp. EDN134. IOP Conf Ser Earth Environ Sci. 2020;572:012024. doi:10.1088/1755-1315/572/1/012024
21. Anita SH, Ardianti FC, Oktaviani M, Sari FP, Nurhayat OD, Ramadhan KP, et al. Immobilization of laccase from Trametes hirsuta EDN 082 in light expanded clay aggregate for decolorization of Remazol Brilliant Blue R dye. Bioresour Technol Rep. 2020;12:100602. doi:10.1016/j.biteb.2020.100602
22. Yanto DHY, Guntoro MA, Nurhayat OD, Anita SH, Oktaviani M, Ramadhan KP, et al. Biodegradation and biodetoxification of batik dye wastewater by laccase from Trametes hirsuta EDN 082 immobilised on light expanded clay aggregate. 3 Biotech. 2021;11(5):247. doi:10.1007/s13205-021-02806-8
23. Noviardi H, Himawan HC, Anggraeni R. Formulasi dan aktivitas antibakteri sediaan gel hand sanitizer dari ekstrak etanol biji mangga harum manis (Mangifera indica L.) terhadap Escherichia coli dan Staphylococcus aureus. J Farmamedika (Pharmamedica J). 2018;3(1):1–9. doi:10.47219/ath.v3i1.20
24. Radji M, Suryadi H. Uji Efektivitas Antimikroba Beberapa Merek Dagang Pembersih Tangan Antiseptik. Pharm Sci Res. 2007;4(1):1–6.doi:10.7454/psr.v4i1.3408
25. Wolfe MK, Lantagne DS. A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens. J Vis Exp. 2017;124:55604. doi:10.3791/55604
26. Fallica F, Leonardi C, Toscano V, Santonocito D, Leonardi P, Puglia C. Assessment of Alcohol-Based Hand Sanitizers for Long-Term Use, Formulated with Addition of Natural Ingredients in Comparison to WHO Formulation 1. Pharmaceutics. 2021;13(4):571. doi:10.3390/pharmaceutics13040571
27. Dolorosa MT, Nurjanah, Purwaningsih S, Anwar E. Utilization of Kappaphycus alvarezii and Sargassum plagyophyllum from Banten as cosmetic creams. IOP Conf Ser Earth Environ Sci. 2020;404:012008. doi:10.1088/1755-1315/404/1/012008
28. Binder L, Mazál J, Petz R, Klang V, Valenta C. The role of viscosity on skin penetration from cellulose ether-based hydrogels. Skin Res Technol. 2019;25(5):725-34. doi:10.1111/srt.12709
29. Kulawik-Pióro A, Miastkowska M. Polymeric Gels and Their Application in the Treatment of Psoriasis Vulgaris: A Review. Int J Mol Sci. 2021;22(10):5124. doi:10.3390/ijms22105124
30. Booq RY, Alshehri AA, Almughem FA, Zaidan NM, Aburayan WS, Bakr AA, et al. Formulation and evaluation of alcohol-free hand sanitizer gels to prevent the spread of infections during pandemics. Int J Environ Res Public Health. 2021;18(12):6252. doi:10.3390/ijerph18126252
31. Febriani A, Syafriana V, Afriyando H, Djuhariah YS. The utilization of oil palm leaves (Elaeis guineensis Jacq.) waste as an antibacterial solid bar soap. IOP Conf Ser Earth Environ Sci. 2020;572:012038. doi:10.1088/1755-1315/572/1/012038
32. Malanovic N, Lohner K. Antimicrobial Peptides Targeting Gram-Positive Bacteria. Pharmaceuticals. 2016;9(3):59. doi:10.3390/ph9030059
33. Syafriana V, Febriani A, Suyatno S, Nurfitri N, Hamida F. Antimicrobial Activity of Ethanolic Extract of Sempur (Dillenia suffruticosa (Griff.) Martelli) Leaves against Pathogenic Microorganisms. Borneo J Pharm. 2021;4(2):135–44. doi:10.33084/bjop.v4i2.1870
34. Wijaya JI. Formulasi Sediaan Gel Hand Sanitizer dengan Bahan Aktif Triklosan 1,5% dan 2%. Calyptra J Ilmiah Mahasiswa Univ Surabaya. 2013;2(1):1–14.
35. Weatherly LM, Gosse JA. Triclosan exposure, transformation, and human health effects. J Toxicol Environ Health B Crit Rev. 2017;20(8):447-69. doi:10.1080/10937404.2017.1399306
36. Kryscio DR, Sathe PM, Lionberger R, Yu L, Bell MA, Jay M, et al. Spreadability measurements to assess structural equivalence (Q3) of topical formulations--a technical note. AAPS PharmSciTech. 2008;9(1):84-6. doi:10.1208/s12249-007-9009-5
37. Basiak E, Lenart A, Debeaufort F. How Glycerol and Water Contents Affect the Structural and Functional Properties of Starch-Based Edible Films. Polymers. 2018;10(4):412. doi:10.3390/polym10040412
Authors
1.
Anita SH, Asishe A, Syafriana V, Febriani A, Zulfiana D, Oktaviani M, Nurhayat OD, Yanto DHY. Hand Sanitizer Gel Formulation with Laccase Enzyme as an Antibacterial Against Staphylococcus aureus and Escherichia coli. Borneo J Pharm [Internet]. 2022Nov.30 [cited 2025Apr.16];5(4):375-83. Available from: https://journal.umpr.ac.id/index.php/bjop/article/view/3683
Copyright (c) 2022 Sita Heris Anita, Asishe Asishe, Vilya Syafriana, Amelia Febriani, Deni Zulfiana, Maulida Oktaviani, Oktan Dwi Nurhayat, Dede Heri Yuli Yanto
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