Аннотация

Allium sativum L. protease still remains largely understudied although new varieties of garlic appear quite often, e.g., lanang garlic. This study tested the antibacterial effect of garlic and the effectiveness of various A. sativum proteases as meat tenderizers.
The research involved powder extracts of four varieties of A. sativum: kating, lanang, black garlic, and sin-chung. The degradation kinetics was defined based on the Lineweaver-Burk equation. The degradation zones were measured using sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS-PAGE). Scan electron microscopy served to test the changes in meat connective tissue.
Lanang demonstrated the largest inhibition zones against Escherichia coli (9.75 ± 0.15 mm) and Staphylococcus aureus (1.04 mm). Sin-chung protease degraded beef protein with the highest V_max of 0.1818 μg/μL/min at 10–22 KDa (small peptide, troponin C, and troponin I), 25–40 KDa (myosin light chain, troponin T, α- and β-tropomyosin, actin), and 100–140 KDa (protein C). The same garlic variety degraded mutton meat protein at 10–17 KDa (small peptide) and 25–40 KDa (myosin light chain, troponin T, α- and β-tropomyosin, actin) with V_max of 0.1135 μg/μL/min.
All four A. sativum proteases proved to be quite effective meat tenderizers.

Ключевые слова

Allium sativum protease, lanang garlic, kating garlic, black garlic, sin-chung garlic

СПИСОК ЛИТЕРАТУРЫ

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2. Chadha J, Harjai K, Chhibber S. Repurposing phytochemicals as anti‐virulent agents to attenuate quorum sensing‐regulated virulence factors and biofilm formation in Pseudomonas aeruginosa. Microbial Biotechnology. 2022;15(6):1695–1718. https://doi.org/10.1111/1751-7915.13981
3. Hong TT, Dat TTH, Hoa NP, Dung TTK, Huyen VTT, Bui LM, et al. Expression and characterization of a new serine protease inhibitory protein in Escherichia coli. Biomedical Research and Therapy. 2020;7(2):3633–3644. https://doi.org/10.15419/bmrat.v7i2.590
4. Rodrigues C, Percival SS. Immunomodulatory effects of glutathione, garlic derivatives, and hydrogen sulfide. Nutrients. 2019;11(2). https://doi.org/10.3390/nu11020295
5. Winiarczyk K, Gębura J. Activity of selected hydrolytic enzymes in Allium sativum L. anthers. Plant Physiology and Biochemistry. 2016;102:37–42. https://doi.org/10.1016/j.plaphy.2016.02.018
6. Ain NU, Riaz S, Abrar S, Ahmad M, Khan Z, Hafiz S, et al. Horizontal gene transfer and antibacterial effect of Allium sativum (garlic) on methicillin-resistant Staphylococcus aureus. Annals of Pakistan Institute of Medical Sciences. 2017;13(2):186–190.
7. Chtita S, Belhassan A, Bakhouch M, Taourati AI, Aouidate A, Belaidi S, et al. QSAR study of unsymmetrical aromatic disulfides as potent avian SARS-CoV main protease inhibitors using quantum chemical descriptors and statistical methods. Chemometrics and Intelligent Laboratory Systems. 2021;210. https://doi.org/10.1016/j.chemolab.2021.104266
8. Ismail RM, Saleh AHA, Ali KS. GC-MS analysis and antibacterial activity of garlic extract with antibiotic. Journal of Medicinal Plants Studies. 2020;8(1):26–30.
9. Kshirsagar MM, Dodamani AS, Karibasappa GN, Vishwakarma PK, Vathar JB, Sonawane KR, et al. Antibacterial activity of garlic extract on cariogenic bacteria: An in vitro study. AYU. 2018;39(3):165–168.
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12. Boutasknit A, Ait-Rahou Y, Anli M, Ait-El-Mokhtar M, Ben-Laouane R, Meddich A. Improvement of garlic growth, physiology, biochemical traits, and soil fertility by Rhizophagus irregularis and compost. Gesunde Pflanzen. 2021;73:149–160. https://doi.org/10.1007/s10343-020-00533-3
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18. Sumiyoshi H, Wargovich MJ. Chemoprevention of 1, 2-dimethylhydrazine-induced colon cancer in mice by naturally occurring organosulfur compounds. Cancer Research. 1990;50(16):5084–5087.
19. Cao Y, Gu W, Zhang J, Chu Y, Ye X, Hu Y, et al. Effects of chitosan, aqueous extract of ginger, onion and garlic on quality and shelf life of stewed-pork during refrigerated storage. Food Chemistry. 2013;141(3):1655–1660. https://doi.org/10.1016/j.foodchem.2013.04.084
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22. Sharma P, Gayen D. Plant protease as regulator and signaling molecule for enhancing environmental stress-tolerance. Plant Cell Reports. 2021;40:2081–2095. https://doi.org/10.1007/s00299-021-02739-9
23. Al-Hadidy YI, Oleiwi SD, Khalaf AA, Saleh HM. The effectiveness of adding apple cider vinegar and garlic to chicken meat kebabs as an antimicrobial and its role in improving its sensory and physiochemical properties. Kirkuk University Journal for Agricultural Sciences. 2023;14(1):117–130. https://doi.org/10.58928/ku23.14110
24. Igwegbe AO, Kassum AL, Maina FJ, Bristone C, Abubakar F, Imam HO, et al. Effects of sodium citrate and garlic on pH and microbial stability of smoked-dried meat stored at ambient temperatures. International Journal of Environmental Health Research. 2020;79.
25. Tkacz K, Modzelewska-Kapituła M, Petracci M, Zduńczyk W. Improving the quality of sous-vide beef from Holstein-Friesian bulls by different marinades. Meat Science. 2021;182. https://doi.org/10.1016/j.meatsci.2021.108639
26. Budianto B, Arumsari AG, Inayah N, Fatmaningrum, Suparmi A. Comparison of the effectiveness of leaf extracts of Anredera cordifolia, Psidium guajava and Pogostemon cablin on inhibitory power over Escherichia coli bacteria. Vitae. 2021;28(3). https://doi.org/10.17533/udea.vitae.v28n3a345386
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29. Běhalová H, Tremlová B, Kalčáková L, Pospiech M, Dordevic D. Assessment of the effect of secondary fixation on the structure of meat products prepared for scanning electron microscopy. Foods. 2020;9(4. https://doi.org/10.3390/foods9040487
30. Vizcaíno AJ, Galafat A, Sáez MI, Martínez TF, Alarcón FJ. Partial characterization of protease inhibitors of Ulva ohnoi and their effect on digestive proteases of marine fish. Marine Drugs. 2020;18(6). https://doi.org/10.3390/md18060319
31. Ankri S, Miron T, Rabinkov A, Wilchek M, Mirelman D. Allicin from garlic strongly inhibits cysteine proteinases and cytopathic effects of Entamoeba histolytica. Antimicrobial Agents and Chemotherapy. 1997;41(10):2286–2288. https://doi.org/10.1128/AAC.41.10.2286
32. Bar M, Binduga UE, Szychowski KA. Methods of isolation of active substances from garlic (Allium sativum L.) and its impact on the composition and biological properties of garlic extracts. Antioxidants. 2022;11(7). https://doi.org/10.3390/antiox11071345
33. Astruc T. Connective tissue: Structure, function, and influence on meat quality. In: Dikeman M, Devine C, editors. Encyclopedia of Meat Sciences. Academic Press; 2014. pp. 321–328. https://doi.org/10.1016/B978-0-12-384731-7.00186-0
34. Swasdison S, Mayne R. Formation of highly organized skeletal muscle fibers in vitro. Comparison with muscle development in vivo. Journal of Cell Science. 1992;102(3):643–652. https://doi.org/10.1242/jcs.102.3.643
35. Batiha GE-S, Beshbishy AM, Wasef LG, Elewa YHA, Al-Sagan AA, Abd El-Hack ME, et al. Chemical constituents and pharmacological activities of garlic (Allium sativum L.): A review. Nutrients. 2020;12(3). https://doi.org/10.3390/nu12030872
36. Chadha J, Harjai K, Chhibber S. Repurposing phytochemicals as anti‐virulent agents to attenuate quorum sensing‐regulated virulence factors and biofilm formation in Pseudomonas aeruginosa. Microbial Biotechnology. 2022;15(6):1695–1718. https://doi.org/10.1111/1751-7915.13981
37. Hong TT, Dat TTH, Hoa NP, Dung TTK, Huyen VTT, Bui LM, et al. Expression and characterization of a new serine protease inhibitory protein in Escherichia coli. Biomedical Research and Therapy. 2020;7(2):3633–3644. https://doi.org/10.15419/bmrat.v7i2.590
38. Rodrigues C, Percival SS. Immunomodulatory effects of glutathione, garlic derivatives, and hydrogen sulfide. Nutrients. 2019;11(2). https://doi.org/10.3390/nu11020295
39. Winiarczyk K, Gębura J. Activity of selected hydrolytic enzymes in Allium sativum L. anthers. Plant Physiology and Biochemistry. 2016;102:37–42. https://doi.org/10.1016/j.plaphy.2016.02.018
40. Ain NU, Riaz S, Abrar S, Ahmad M, Khan Z, Hafiz S, et al. Horizontal gene transfer and antibacterial effect of Allium sativum (garlic) on methicillin-resistant Staphylococcus aureus. Annals of Pakistan Institute of Medical Sciences. 2017;13(2):186–190.
41. Chtita S, Belhassan A, Bakhouch M, Taourati AI, Aouidate A, Belaidi S, et al. QSAR study of unsymmetrical aromatic disulfides as potent avian SARS-CoV main protease inhibitors using quantum chemical descriptors and statistical methods. Chemometrics and Intelligent Laboratory Systems. 2021;210. https://doi.org/10.1016/j.chemolab.2021.104266
42. Ismail RM, Saleh AHA, Ali KS. GC-MS analysis and antibacterial activity of garlic extract with antibiotic. Journal of Medicinal Plants Studies. 2020;8(1):26–30.
43. Kshirsagar MM, Dodamani AS, Karibasappa GN, Vishwakarma PK, Vathar JB, Sonawane KR, et al. Antibacterial activity of garlic extract on cariogenic bacteria: An in vitro study. AYU. 2018;39(3):165–168.
44. Parthipan P, Elumalai P, Narenkumar J, Machuca LL, Murugan K, Karthikeyan OP, et al. Allium sativum (garlic extract) as a green corrosion inhibitor with biocidal properties for the control of MIC in carbon steel and stainless steel in oilfield environments. International Biodeterioration and Biodegradation. 2018;132:66–73. https://doi.org/10.1016/j.ibiod.2018.05.005
45. Piletti R, Zanetti M, Jung G, de Mello JMM, Dalcanton F, Soares C, et al. Microencapsulation of garlic oil by β-cyclodextrin as a thermal protection method for antibacterial action. Materials Science and Engineering: C. 2019;94:139–149. https://doi.org/10.1016/j.msec.2018.09.037
46. Boutasknit A, Ait-Rahou Y, Anli M, Ait-El-Mokhtar M, Ben-Laouane R, Meddich A. Improvement of garlic growth, physiology, biochemical traits, and soil fertility by Rhizophagus irregularis and compost. Gesunde Pflanzen. 2021;73:149–160. https://doi.org/10.1007/s10343-020-00533-3
47. Mengesha W, Tesfaye A, Djene M. Evaluation of fungicides on the control of garlic rust (Pucinnia allii) in Eastern Ethiopia. International Journal of Emerging Technology and Advanced Engineering. 2016;6(1):27–33.
48. Khubber S, Hashemifesharaki R, Mohammadi M, Gharibzahedi SMT. Garlic (Allium sativum L.): A potential unique therapeutic food rich in organosulfur and flavonoid compounds to fight with COVID-19. Nutrition Journal. 2020;19. https://doi.org/10.1186/s12937-020-00643-8
49. Gorinstein S, Jastrzebski Z, Namiesnik J, Leontowicz H, Leontowicz M, Trakhtenberg S. The atherosclerotic heart disease and protecting properties of garlic: contemporary data. Molecular Nutrition and Food Research. 2007;51(11):1365–1381. https://doi.org/https://doi.org/10.1002/mnfr.200700064
50. Kritchevsky D. The effect of dietary garlic on the development of cardiovascular disease. Trends in Food Science and Technology. 1991;2:141–144. https://doi.org/10.1016/0924-2244(91)90658-6
51. Devrim E, Durak I. Is garlic a promising food for benign prostatic hyperplasia and prostate cancer? Molecular Nutrition and Food Research. 2007;51(11):1319–1323. https://doi.org/10.1002/mnfr.200600302
52. Sumiyoshi H, Wargovich MJ. Chemoprevention of 1, 2-dimethylhydrazine-induced colon cancer in mice by naturally occurring organosulfur compounds. Cancer Research. 1990;50(16):5084–5087.
53. Cao Y, Gu W, Zhang J, Chu Y, Ye X, Hu Y, et al. Effects of chitosan, aqueous extract of ginger, onion and garlic on quality and shelf life of stewed-pork during refrigerated storage. Food Chemistry. 2013;141(3):1655–1660. https://doi.org/10.1016/j.foodchem.2013.04.084
54. Aoki-Shioi N, Nagai Y, Deshimaru M, Terada S. Precursor genes of Bowman-Birk-type serine proteinase inhibitors comprise multiple inhibitory domains to promote diversity. Biochimica et Biophysica Acta (BBA) – General Subjects. 2023;1867(1). https://doi.org/10.1016/j.bbagen.2022.130248
55. Guo R, Zhao J, Wang X, Guo C, Li Z, Wang Y, et al. Constitutive expression of a grape aspartic protease gene in transgenic Arabidopsis confers osmotic stress tolerance. Plant Cell, Tissue and Organ Culture. 2015;121:275–287. https://doi.org/10.1007/s11240-014-0699-6
56. Sharma P, Gayen D. Plant protease as regulator and signaling molecule for enhancing environmental stress-tolerance. Plant Cell Reports. 2021;40:2081–2095. https://doi.org/10.1007/s00299-021-02739-9
57. Al-Hadidy YI, Oleiwi SD, Khalaf AA, Saleh HM. The effectiveness of adding apple cider vinegar and garlic to chicken meat kebabs as an antimicrobial and its role in improving its sensory and physiochemical properties. Kirkuk University Journal for Agricultural Sciences. 2023;14(1):117–130. https://doi.org/10.58928/ku23.14110
58. Igwegbe AO, Kassum AL, Maina FJ, Bristone C, Abubakar F, Imam HO, et al. Effects of sodium citrate and garlic on pH and microbial stability of smoked-dried meat stored at ambient temperatures. International Journal of Environmental Health Research. 2020;79.
59. Tkacz K, Modzelewska-Kapituła M, Petracci M, Zduńczyk W. Improving the quality of sous-vide beef from Holstein-Friesian bulls by different marinades. Meat Science. 2021;182. https://doi.org/10.1016/j.meatsci.2021.108639
60. Budianto B, Arumsari AG, Inayah N, Fatmaningrum, Suparmi A. Comparison of the effectiveness of leaf extracts of Anredera cordifolia, Psidium guajava and Pogostemon cablin on inhibitory power over Escherichia coli bacteria. Vitae. 2021;28(3). https://doi.org/10.17533/udea.vitae.v28n3a345386
61. Manzoor M, Singh J, Ray A, Gani A. Recent advances in analysis of food proteins. In: Gani A, Ashwar BA, editors. Food biopolymers: Structural, functional and nutraceutical properties. Cham: Springer; 2021. pp. 269–298. https://doi.org/10.1007/978-3-030-27061-2_12
62. Official Methods of Analysis of the Association of Official Analytical Chemists. 18th ed. Washington: Association of Official Analytical Chemists Washington; 2010.
63. Běhalová H, Tremlová B, Kalčáková L, Pospiech M, Dordevic D. Assessment of the effect of secondary fixation on the structure of meat products prepared for scanning electron microscopy. Foods. 2020;9(4. https://doi.org/10.3390/foods9040487
64. Vizcaíno AJ, Galafat A, Sáez MI, Martínez TF, Alarcón FJ. Partial characterization of protease inhibitors of Ulva ohnoi and their effect on digestive proteases of marine fish. Marine Drugs. 2020;18(6). https://doi.org/10.3390/md18060319
65. Ankri S, Miron T, Rabinkov A, Wilchek M, Mirelman D. Allicin from garlic strongly inhibits cysteine proteinases and cytopathic effects of Entamoeba histolytica. Antimicrobial Agents and Chemotherapy. 1997;41(10):2286–2288. https://doi.org/10.1128/AAC.41.10.2286
66. Bar M, Binduga UE, Szychowski KA. Methods of isolation of active substances from garlic (Allium sativum L.) and its impact on the composition and biological properties of garlic extracts. Antioxidants. 2022;11(7). https://doi.org/10.3390/antiox11071345
67. Astruc T. Connective tissue: Structure, function, and influence on meat quality. In: Dikeman M, Devine C, editors. Encyclopedia of Meat Sciences. Academic Press; 2014. pp. 321–328. https://doi.org/10.1016/B978-0-12-384731-7.00186-0
68. Swasdison S, Mayne R. Formation of highly organized skeletal muscle fibers in vitro. Comparison with muscle development in vivo. Journal of Cell Science. 1992;102(3):643–652. https://doi.org/10.1242/jcs.102.3.643