Optimization of Soy Protein Hydrolysis with Papain Enzyme and Its Cytotoxic Activity Against MCF-7 Cancer Cells

Sandra Hermanto, Syadzwina Nurdzakiyyah Ahmad, Tarso Rudiana

Abstrak


Soybean is a source of vegetable protein that is rich in nutrients and a source of producing anticancer bioactive peptides. This study aims to determine the optimum conditions for hydrolysis of soybean protein with papain enzyme and their cytotoxicity against the MCF-7 cells. Soybean protein was isolated using an acid precipitation technique. Then, the protein isolate was hydrolyzed using papain enzyme with variations of papain concentration 0,5; 1; and 5% (v/v) and variations of incubation time 0, 1, 2, 3, and 4 hours at 50 °C. The hydrolysates were tested for their degree of hydrolysis (%DH), molecular weight profile using SDS-PAGE, and cytotoxicity against the MCF-7 cells through an in-vitro assay. The most active hydrolysate was fractionated using Sephadex G-15 and characterized by the molecular weight by LCMS/MS. The result showed that the optimum condition for hydrolysis was 1% (v/v) of enzyme concentration and 3 hours of incubation time with a %DH value of 3.01%. Based on the SDS-PAGE result, the hydrolysate had protein bands in a lower range (<25 kDa). That hydrolysate has cytotoxicity with an IC50 value of 1.87 mg/mL, and the molecular weight of its bioactive peptide is 7.70 kDa.

Kata Kunci


bioactive peptide; cytotoxicity; MCF-7 cells; papain; soybean

Teks Lengkap:

PDF (English)

Referensi


World Health Organization (WHO). Breast Cancer [Internet]. 2021 [cited 2021 Oct 20]. Available from: https://www.who.int/news-room/fact-sheets/detail/breast-cancer

Neophytou CM, Trougakos IP, Erin N, Papageorgis P. Apoptosis Deregulation and the Development of Cancer Multi-Drug Resistance. Cancers. 2021; 13(17):4363. https://doi.org/10.3390/cancers13174363

Jia H, Zhou T, Zhu H, Shen L, He P. Quantification of Gly m 5.0101 in Soybean and Soy Products by Liquid Chromatography-Tandem Mass Spectrometry. Molecules. 2018 Dec 25;24(1):68. doi: 10.3390/molecules24010068. PMID: 30585221; PMCID: PMC6337133.

Chatterjee C, Gleddie S, Xiao CW. Soybean Bioactive Peptides and Their Functional Properties. Nutrients. 2018 Sep 1;10(9):1211. doi: 10.3390/nu10091211. PMID: 30200502; PMCID: PMC6164536.

Alves de Souza SM, Hernández-Ledesma B, de Souza TLF. Lunasin as a Promising Plant-Derived Peptide for Cancer Therapy. International Journal of Molecular Sciences. 2022; 23(17):9548. https://doi.org/10.3390/ijms23179548

Hermanto S, Octavio A, Azrifitria, Kusumaningrum S. The HMG-CoA Reductase Inhibitor Activities of Soy Protein Hydrolysates from Papain Hydrolysis. Molekul. 2021;16(2):145–55. https://doi.org/http://dx.doi.org/10.20884/1.jm.2021.16.2.724

Sitanggang AB, Putri JE, Palupi NS, Hatzakis E, Syamsir E, Budijanto S. Enzymatic Preparation of Bioactive Peptides Exhibiting Ace Inhibitory Activity from Soybean and Velvet Bean: A Systematic Review. Molecules. 2021;26(13). https://doi.org/10.3390/molecules26133822

Rayaprolu SJ, Hettiarachchy NS, Horax R, Phillips GK, Mahendran M, Chen P. Soybean Peptide Fractions Inhibit Human Blood, Breast and Prostate Cancer Cell Proliferation. J Food Sci Technol. 2017;54(1):38–44. https://doi.org/10.1007/s13197-016-2426-2

Hsie CH, Wang TY, Tung BC, Liu HP, Yeh LT, Hsu KC. The Hydrolytic Peptides of Soybean Protein Induce Cell Cycle Arrest and Apoptosis on Human Oral Cancer Cell Line HSC-3. Molecules. 2022 Apr 29;27(9):2839. doi: 10.3390/molecules27092839. PMID: 35566188; PMCID: PMC9101267.

Bradford MM. A Rapid and Sensitive Method for the Quantitation Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Anal Biochem. 1976;72:248–54. https://doi.org/10.1006/abio.1976.9999

Laemmli UK. Cleavage of Structural Proteins during The Assembly of The Head of Bacteriophage T4. Nature. 1970;227(5259):680–5. https://doi.org/https://doi.org/10.1038/227680a0

Leni G, Soetemans L, Caligiani A, Sforza S, Bastiaens L. Degree of Hydrolysis Affects the Techno-Functional Properties of Lesser Mealworm Protein Hydrolysates. Foods. 2020 Mar 25;9(4):381. doi: 10.3390/foods9040381. PMID: 32218377; PMCID: PMC7230224.

Islam M, Huang Y, Islam S, Fan B, Tong L, Wang F. Influence of the Degree of Hydrolysis on Functional Properties and Antioxidant Activity of Enzymatic Soybean Protein Hydrolysates. Molecules. 2022; 27(18):6110. https://doi.org/10.3390/molecules27186110

Vitolo M. Brief Review on Enzyme Activity. World J Pharm Res. 2020;9(2):60–76. https://doi.org/10.20959/wjpr20202-16660

Sirison J, Matsumiya K, Samoto M, Hidaka H, Kouno M & Matsumura Y, Solubility of soy lipophilic proteins: comparison with other soy protein fractions, Bioscience, Biotechnology, and Biochemistry, 2017; 81:4, 790-802, DOI: 10.1080/09168451.2017.1282808

Guan X, Zhong X, Lu Y, Du X, Jia R, Li H, Zhang M. Changes of Soybean Protein during Tofu Processing. Foods. 2021; 10(7):1594. https://doi.org/10.3390/foods10071594

Wong DWS. Mechanism and Theory in Food Chemistry. Second Edi. California: Springer; 2018. https://doi.org/10.1007/978-3-319-50766-8

Chen Z, Li W, Santhanam RK, Wang C, Gao X, Chen Y, et al. Bioactive Peptide with Antioxidant and Anticancer Activities from Black Soybean [Glycine max (L.) Merr.] Byproduct: Isolation, Identification and Molecular Docking Study. Eur Food Res Technol. 2019;245(3):677–89. http[1]. World Health Organization (WHO). Breast Cancer [Internet]. 2021 [cited 2021 Oct 20]. Available from: https://www.who.int/news-room/fact-sheets/detail/breast-cancer

Galvez AF, De Lumen BO. A soybean cDNA encoding a chromatin-binding peptide inhibits mitosis of mammalian cells. Nat Biotechnol. 1999;17:495–500. https://doi.org/https://doi.org/10.1038/8676

Zhu Y., Anticancer Effects of Soybean Bioactive Components and Anti-inflammatory Activities of the Soybean Peptide Lunasin. 2018. (Thèse de doctorat en anglais). Gembloux, Belgique, Gembloux Agro-Bio Tech, Universitéde Liège

Ezekiel A., Florence M. Papain, a plant enzyme of biological importance: A review. Am J Biochem. Biotechnol. 2012; 8:99–104

Lu J, Xu H, Xia J, Ma J, Xu J, Li Y, Feng J. D- and Unnatural Amino Acid Substituted Antimicrobial Peptides With Improved Proteolytic Resistance and Their Proteolytic Degradation Characteristics. Front Microbiol. 2020 Nov 12;11:563030. doi: 10.3389/fmicb.2020.563030. PMID: 33281761; PMCID: PMC7688903.

Sun H, Qiao B, Choi W, Hampu N, Naneki C. McCallum, Matthew P. Thompson, Oktawiec J, Weigand S, Ebrahim UM, de la Cruz MO, and Gianneschi NC, Origin of Proteolytic Stability of Peptide-Brush Polymers as Globular Proteomimetics ACS Central Science 2021 7 (12), 2063-2072 DOI: 10.1021/acscentsci.1c01149




DOI: https://doi.org/10.25077/jsfk.10.1.137-144.2023

Article Metrics

Abstract view : 326 times
PDF (English) view/download : 198 times



JSFK (Jurnal Sains Farmasi & Klinis) (J Sains Farm Klin) | p-ISSN: 2407-7062 | e-ISSN: 2442-5435

Diterbitkan oleh Fakultas Farmasi Universitas Andalas bekerjasama dengan Ikatan Apoteker Indonesia - Daerah Sumatera Barat 

      

 JSFK is licensed under Creative Commons Attribution-ShareAlike 4.0 International License.