ANGIOTENSIN-I-CONVERTING ENZYME INHIBITORY AND ANTIOXIDANT ACTIVITY OF TRYPTIC SPIRULINA PLATENSIS PROTEIN HYDROLYSATES: EFFECT OF HYDROLYSIS AND IN VITRO GASTROINTESTINAL DIGESTION

Yıl 2018, Cilt: 13 Sayı: 3, 151 - 162, 24.07.2018

Aysun Yücetepe , Kadriye Nur Kasapoğlu Beraat Özçelik

Öz

In
this study, protein hydrolysates derived from Spirulina platensis protein (SPPHs) using trypsin
were investigated in terms of angiotensin I-converting enzyme (ACE) inhibitory
activity, antioxidant activity and total phenolic content (TPC) and subjected
to an in vitro digestion model using
human gastric and duodenal fluids. Moreover, the effects of hydrolysis time and
enzyme/substrate (E/S) ratio on the degree of hydrolysis (DH) of the
hydrolysates were determined. The maximum DH (%) was found as 25.03±0.89% with
the combination of E/S ratio of 3:100, hydrolysis time of 8 hours (p<0.05).
The highest ACE inhibitory activity value was observed as 21.79±1.52% for
initial SPPHs, prepared within the hydrolysis conditions of E/S ratio of 3:100
and hydrolysis time of 8 h. In general, the increase in E/S ratio and
hydrolysis time resulted in an increase in the DH and in an improved ACE
inhibitory activity of both initial and the GI digested samples (p<0.05).
After digestion by pepsin, TPC of the digests was in the range of 28.87±0.32
and 40.28±1.05mg caffeic acid equivalent/g dry weight. However, further
digestion by pancreatin led TPC of the final GI digest between 19.85±1.24 and
29.00±1.00mg caffeic acid equivalent/g dry weight. Moreover, the antioxidant
activity of further digested SPPHs by gastric and intestinal proteases remained
generally stable after in vitro treatment.

Anahtar Kelimeler

Angiotensin I-converting Enzyme, Antioxidant Activity, Bioaccessibility, Enzymatic Hydrolysis, Spirulina platensis

ANGIOTENSIN-I-CONVERTING ENZYME INHIBITORY AND ANTIOXIDANT ACTIVITY OF TRYPTIC SPIRULINA PLATENSIS PROTEIN HYDROLYSATES: EFFECT OF HYDROLYSIS AND IN VITRO GASTROINTESTINAL DIGESTION

Öz

In this study, protein hydrolysates derived from Spirulina platensis protein (SPPHs) using trypsin were investigated in terms of angiotensin I-converting enzyme (ACE) inhibitory activity, antioxidant activity and total phenolic content (TPC) and subjected to an in vitro digestion model using human gastric and duodenal fluids. Moreover, the effects of hydrolysis time and enzyme/substrate (E/S) ratio on the degree of hydrolysis (DH) of the hydrolysates were determined. The maximum DH (%) was found as 25.03±0.89% with the combination of E/S ratio of 3:100, hydrolysis time of 8 hours (p<0.05). The highest ACE inhibitory activity value was observed as 21.79±1.52% for initial SPPHs, prepared within the hydrolysis conditions of E/S ratio of 3:100 and hydrolysis time of 8 h. In general, the increase in E/S ratio and hydrolysis time resulted in an increase in the DH and in an improved ACE inhibitory activity of both initial and the GI digested samples (p<0.05). After digestion by pepsin, TPC of the digests was in the range of 28.87±0.32 and 40.28±1.05mg caffeic acid equivalent/g dry weight. However, further digestion by pancreatin led TPC of the final GI digest between 19.85±1.24 and 29.00±1.00mg caffeic acid equivalent/g dry weight. Moreover, the antioxidant activity of further digested SPPHs by gastric and intestinal proteases remained generally stable after in vitro treatment.

Anahtar Kelimeler

Angiotensin I-converting Enzyme, Antioxidant Activity, Bioaccessibility, Enzymatic Hydrolysis, Spirulina platensis

Kaynakça

• 1. Wang, L., Pan, B., Sheng, J., Xu, J., and Hu, Q., (2007). Antioxidant Activity of Spirulina Platensis Extracts By Supercritical Carbon Dioxide Extraction. Food Chemistry. 105(1): p. 36-41.
• 2. Lupatini, A.L., Colla, L.M., Canan, C., and Colla, E., (2016). Potential Application of Microalgae Spirulina Platensis as A Protein Source. Journal of the Science of Food and Agriculture. 97(3):724-732.
• 3. Zhang, L., Chen, L., Wang, J., Chen, Y., Gao, X., Zhang, Z., and Liu, T., (2015). Attached Cultivation for Improving The Biomass Productivity of Spirulina Platensis. Bioresource technology. 181:136-142.
• 4. Gad, A.S., Khadrawy, Y.A., El-Nekeety, A.A., Mohamed, S.R., Hassan, N.S., and Abdel-Wahhab, M.A., (2011). Antioxidant Activity and Hepatoprotective Effects of Whey Protein and Spirulina in Rats. Nutrition. 27(5):582-589.
• 5. Kim, S.K. and Kang, K.H., (2011). Medicinal Effects of Peptides from Marine Microalgae. Advances in food and nutrition research. 64:313-323.
• 6. Benelhadj, S., Gharsallaoui, A., Degraeve, P., Attia H. and Ghorbel, D. (2016). Effect of Ph on The Functional Properties of Arthrospira (Spirulina) Platensis Protein Isolate. Food Chemistry. 194:1056- 1063.
• 7. Fradique, M., Batista, A.P., Nunes, M.C., Gouveia, L., Bandarra N.M., and Raymundo, A., (2010). Incorporation of Chlorella Vulgaris and Spirulina Maxima Biomass in Pasta Products. Part 1: Preparation and evaluation. Journal of the Science of Food and Agriculture. 90(10):1656-1664.
• 8. Di Bernardini, R., Harnedy, P., Bolton, D., Kerry, J., O’Neill, E., Mullen, A.M., and Hayes, M., (2011). Antioxidant and Antimicrobial Peptidic Hydrolysates from Muscle Protein Sources and By-Products. Food Chemistry. 124(4):1296-1307.
• 9. Carrasco-Castilla, J., Hernández-Álvarez, A.J., Jiménez-Martínez, C., Jacinto-Hernández, C., Alaiz, M., Girón-Calle, J., Vioque, J., and Dávila-Ortiz, G., (2012). Antioxidant and Metal Chelating Activities of Phaseolus Vulgaris L. Var. Jamapa Protein Isolates, Phaseolin and Lectin Hydrolysates. Food Chemistry. 131(4):1157-1164.
• 10. Hartmann, R. and Meisel, H., (2007). Food-Derived Peptides with Biological Activity: From Research to Food Applications. Current opinion in biotechnology. 18(2):163-169.
• 11. Jamdar, S., Rajalakshmi, V., Pednekar, M., Juan, F., Yardi, V., and Sharma, A., (2010). Influence of Degree of Hydrolysis on Functional Properties, Antioxidant Activity and ACE Inhibitory Activity of Peanut Protein Hydrolysate. Food Chemistry. 121(1):178-184.
• 12. Ahn, C.B., Jeon, Y.J. Kim, Y.T., and Je, J.Y., (2012). Angiotensin I Converting Enzyme (ACE) Inhibitory Peptides from Salmon Byproduct Protein Hydrolysate by Alcalase Hydrolysis. Process Biochemistry. 47(12):2240-2245.
• 13. Udenigwe, C.C., (2014). Bioinformatics Approaches, Prospects and Challenges of Food Bioactive Peptide Research. Trends in Food Science & Technology. 36(2):137-143.
• 14. Korhonen, H., (2009). Milk-Derived Bioactive Peptides: From Science to Applications. Journal of Functional Foods. 1(2):177-187.
• 15. Power, O., Jakeman, P., and FitzGerald, R., (2013). Antioxidative Peptides: Enzymatic Production, In Vitro and In Vivo Antioxidant Activity and Potential Applications of Milk-Derived Antioxidative Peptides. Amino Acids. 44(3):797-820.
• 16. Yu, J., Hu, Y., Xue, M., Dun, Y., Li, S., Peng, N., Liang, Y., and Zhao, S., (2016). Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysate of Spirulina Platensis. Journal of Microbiolgy and Biotechnolgy. 26(7):1216-1223.
• 17. Sun, Y., Chang, R., Li, Q., and Li, B., (2016). Isolation and Characterization of an Antibacterial Peptide from Protein Hydrolysates of Spirulina Platensis. European Food Research and Technology. 242(5):685-692.
• 18. Pan, H., She, X., Wu, H., Ma, J., Ren, D., and Lu, J., (2015). Long-Term Regulation of The Local Renin–Angiotensin System in The Myocardium of Spontaneously Hypertensive Rats By Feeding Bioactive Peptides Derived From Spirulina Platensis. Journal of agricultural and food chemistry. 63(35): p. 7765-7774.
• 19. Lu, J., Ren, D.F., Xue, Y.L., Sawano, Y., Miyakawa, T., and Tanokura, M., (2010). Isolation of an Antihypertensive Peptide From Alcalase Digest of Spirulina Platensis. Journal of agricultural and food chemistry. 58(12):7166-7171.
• 20. He, H.L., Chen, X.L. Wu, H., Sun, C.Y. Zhang, Y.Z., and Zhou, B.C., (2007). High Throughput and Rapid Screening of Marine Protein Hydrolysates Enriched in Peptides with Angiotensin-I-Converting Enzyme Inhibitory Activity by Capillary Electrophoresis. Bioresource Technology. 98(18):3499-3505.
• 21. Wang, Z. and Zhang, X., (2016). Characterization and Antitumor Activity of Protein Hydrolysates from Arthrospira Platensis (Spirulina Platensis) Using Two-Step Hydrolysis. Journal of Applied Phycology. 28(6):3379-3385.
• 22. Wang, Z. and Zhang, X., (2017). Isolation and Identification of Anti‐Proliferative Peptides from Spirulina Platensis Using Three‐Step Hydrolysis. Journal of the Science of Food and Agriculture. 97(3):918-922.
• 23. Kim, N.H., Jung, S.H., Kim, J., Kim, S.H., Ahn, H.J., and Song, K.B., (2014). Purification of an Iron-Chelating Peptide from Spirulina Protein Hydrolysates. Journal of the Korean Society for Applied Biological Chemistry. 57(1):91-95.
• 24. Adjonu, R., Doran, G., Torley, P., and Agboola, S., (2013). Screening of Whey Protein Isolate Hydrolysates for Their Dual Functionality: Influence of Heat Pre-Treatment and Enzyme Specificity. Food chemistry. 136(3):1435-1443.
• 25. Kristinsson, H.G. and Rasco, B.A., (2000). Fish Protein Hydrolysates: Production, Biochemical, and Functional Properties. Critical Reviews in Food Science and Nutrition. 40(1):43-81.
• 26. Kishimura, H., Tokuda, Y., Yabe, M., Klomklao, S., Benjakul, S., and Ando, S., (2007). Trypsins from the Pyloric Ceca of Jacopever (Sebastes Schlegelii) and Elkhorn Sculpin (Alcichthys Alcicornis): Isolation and Characterization. Food Chemistry. 100(4):1490-1495.
• 27. Choonpicharn, S., Jaturasitha, S., Rakariyatham, N., Suree, N., and Niamsup, H., (2015). Antioxidant and Antihypertensive Activity of Gelatin Hydrolysate from Nile Tilapia Skin. Journal of food science and technology. 52(5):3134-3139.
• 28. Abedin, M.Z., Karim, A.A., Latiff, A.A., Gan, C.Y., Ghazali, F.C., Barzideh, Z., Ferdosh, S., Akanda, M.J.H., Zzaman, W., and Karim, M.R., (2014). Biochemical and Radical-Scavenging Properties of Sea Cucumber (Stichopus Vastus) Collagen Hydrolysates. Natural product research. 28(16):1302-1305.
• 29. Chatterjee, A., Kanawjia, S., Khetra, Y., and Saini, P., (2015). Discordance Between in Silico and in Vitro Analyses of ACE Inhibitory and Antioxidative Peptides from Mixed Milk Tryptic Whey Protein Hydrolysate. Journal of Food Science and Technology. 52(9):5621- 5630.
• 30. Vo, T.S. and Kim, S.K., (2013). Down-Regulation of Histamine-Induced Endothelial Cell Activation as Potential Anti-Atherosclerotic Activity of Peptides from Spirulina Maxima. European Journal of Pharmaceutical Sciences. 50(2):198-207.
• 31. Mäkinen, S., Johannson, T., Gerd, E.V., Pihlava, J.M., and Pihlanto, A., (2012). Angiotensin I-Converting Enzyme Inhibitory and Antioxidant Properties of Rapeseed Hydrolysates. Journal of Functional Foods. 4(3):575-583.
• 32. Estrada, J.P., Bescos, P.B., and Del Fresno, A.V., (2001). Antioxidant Activity of Different Fractions of Spirulina Platensis Protean Extract. Il farmaco. 56(5):497-500.
• 33. Ling, A.L.M., (2014). Antioxidant Activity, Total Phenolic and Flavonoid Contents of Selected Commercial Seaweeds of Sabah, Malaysia. International Journal of Pharmaceutical and Phytopharmacological Research. 3(3).
• 34. Chanput, W., Theerakulkait, C., and Nakai, S., (2009). Antioxidative Properties of Partially Purified Barley Hordein, Rice Bran Protein Fractions and Their Hydrolysates. Journal of Cereal Science. 49(3):422-428.
• 35. Margot, A., Flaschel, E., and Renken, A., (1997). Empirical Kinetic Models for Tryptic Whey-Protein Hydrolysis. Process Biochemistry. 32(3):217-223.
• 36. Karamac, M., Amarowicz, R., and Kostyra, H., (2002). Effect of Temperature and Enzyme/Substrate Ratio on the Hydrolysis of Pea Protein Isolates by Trypsin. Czech Journal of Food Sciences. 20(1):1-6.
• 37. Vaštag, Ž., Popović, L., Popović, S., Peričin-Starčević, I., and Krimer-Malešević, V., (2013). In Vitro Study on Digestion of Pumpkin Oil Cake Protein Hydrolysate: Evaluation of Impact on Bioactive Properties. International journal of Food Sciences and Nutrition. 64(4):452-460.
• 38. Jadhav, S., Lutz, S., Ghorpade, V., and Salunkhe, D., (1998). Barley: Chemistry and Value-Added Processing. Critical Reviews in Food Science. 38(2):123-171.
• 39. Hausch, F., Shan, L., Santiago, N.A., Gray, G.M., and Khosla, C., (2002). Intestinal Digestive Resistance of Immunodominant Gliadin Peptides. American Journal of Physiology-Gastrointestinal and Liver Physiology. 283(4):G996-G1003.
• 40. Simpson, D.J., (2001). Proteolytic Degradation of Cereal Prolamins—the Problem With Proline. Plant Science. 161(5):825-838.
• 41. Balogh, E., Hegedűs, A., and Stefanovits-Bányai, É., (2010). Application of and Correlation among Antioxidant and Antiradical Assays for Characterizing Antioxidant Capacity of Berries. Scientia horticulturae. 125(3):332-336.
• 42. Apak, R., Güçlü, K., Özyürek, M., and Karademir, S.E., (2004). Novel Total Antioxidant Capacity Index for Dietary Polyphenols And Vitamins C and E, Using Their Cupric Ion Reducing Capability In The Presence of Neocuproine: CUPRAC method. Journal of Agricultural and Food Chemistry. 52(26):7970-7981.
• 43. Önay-Uçar, E., Arda, N., Pekmez, M., Yılmaz, A.M., Böke-Sarıkahya, N., Kırmızıgül, S., and Yalçın, A.S., (2014). Comparison of Antioxidant Capacity, Protein Profile and Carbohydrate Content of Whey Protein Fractions. Food Chemistry. 150:34-40.
• 44. Wiriyaphan, C., Chitsomboon, B. and Yongsawadigul, J. (2012). Antioxidant Activity Of Protein Hydrolysates Derived from Threadfin Bream Surimi Byproducts. Food Chemistry. 132(1): p. 104-111.
• 45. De Castro, R.J.S. and Sato, H.H., (2015). A Response Surface Approach on Optimization of Hydrolysis Parameters for the Production of Egg White Protein Hydrolysates with Antioxidant Activities. Biocatalysis and Agricultural Biotechnology. 4(1):55-62.
• 46. Hsu, K.C., (2010). Purification of Antioxidative Peptides Prepared From Enzymatic Hydrolysates of Tuna Dark Muscle By-Product. Food Chemistry. 122(1):42-48.
• 47. Liu, Q., Kong, B., Xiong, Y.L., and Xia, X., (2010). Antioxidant Activity and Functional Properties of Porcine Plasma Protein Hydrolysate as Influenced by the Degree of Hydrolysis. Food Chemistry. 118(2):403-410.
• 48. Foltz, M., Van Buren, L., Klaffke, W., and Duchateau, G.S., (2009). Modeling of the Relationship between Dipeptide Structure and Dipeptide Stability, Permeability, and ACE Inhibitory Activity. Journal of food science. 74(7):H243-H251.
• 49. You, L., Zhao, M., Regenstein, J.M., and Ren, J., (2010). Changes in the Antioxidant Activity of Loach (Misgurnus Anguillicaudatus) Protein Hydrolysates during a Simulated Gastrointestinal Digestion. Food Chemistry. 120(3):810-816.
• 50. Silva, F., O’Callagahan, Y., O’Brien, N., and Netto, F., (2013). Antioxidant Capacity of Flaxseed Products: The Effect of in Vitro Digestion. Plant Foods for Human Nutrition. 68(1):24-30.
• 51. Ao, J. and Li, B., (2013). Stability and Antioxidative Activities of Casein Peptide Fractions During Simulated Gastrointestinal Digestion in Vitro: Charge Properties of Peptides Affect Digestive Stability. Food Research International. 52(1):334-341.
• 52. Teixeira, B., Pires, C., Nunes, M.L., and Batista, I., (2016). Effect of In Vitro Gastrointestinal Digestion on The Antioxidant Activity of Protein Hydrolysates Prepared from Cape Hake By‐Products. International Journal of Food Science and Technology. 51(12):2528-2536.
• 53. Samaranayaka, A.G. and Li-Chan, E.C., (2011). Food-Derived Peptidic Antioxidants: A Review of Their Production, Assessment, and Potential Applications. Journal of Functional Foods. 3(4):229-254.
• 54. Cian, R.E., Caballero, M.S., Sabbag, N., González, R.J., and Drago, S.R., (2014). Bio-Accessibility of Bioactive Compounds (ACE Inhibitors and Antioxidants) from Extruded Maize Products Added with A Red Seaweed Porphyra Columbina. LWT-Food Science and Technology. 55(1):51-58.
• 55. Cian, R.E., Martínez-Augustin, O., and Drago, S.R., (2012). Bioactive Properties of Peptides Obtained By Enzymatic Hydrolysis from Protein Byproducts of Porphyra Columbina. Food Research International. 49(1):364-372.
• 56. Lu, J., Sawano, Y., Miyakawa, T., Xue, Y.L., Cai, M.Y., Egashira, Y., Ren, D.F., and Tanokura, M., (2010). One-Week Antihypertensive Effect of Ile-Gln-Pro in Spontaneously Hypertensive Rats. Journal of Agricultural and Food Chemistry. 59(2):559-563.
• 57. Sheih, I.C., Fang, T.J., and Wu, T.K., (2009). Isolation and Characterisation of A Novel Angiotensin I-Converting Enzyme (ACE) Inhibitory Peptide From the Algae Protein Waste. Food Chemistry. 115(1):279-284.
• 58. Suetsuna, K. and Chen, J.R., (2001). Identification of Antihypertensive Peptides from Peptic Digest of Two Microalgae, Chlorella Vulgaris and Spirulina Platensis. Marine Biotechnology. 3(4):305-309.
• 59. Suetsuna, K. and Nakano, T. (2000). Identification of an Antihypertensive Peptide from Peptic Digest of Wakame (Undaria Pinnatifida). The Journal of Nutritional Biochemistry. 11(9):450-454.
• 60. Boschin, G., Scigliuolo, G.M., Resta, D., and Arnoldi, A., (2014). ACE- Inhibitory Activity of Enzymatic Protein Hydrolysates from Lupin and Other Legumes. Food Chemistry. 145:34-40.
• 61. Matsui, T., Matsufuji, H., Seki, E., Osajima, K., Nakashima, M., and Osajima, Y., (1993). Inhibition of Angiotensin I-Converting Enzyme By Bacillus Licheniformis Alkaline Protease Hydrolyzates Derived From Sardine Muscle. Bioscience, Biotechnology, and Biochemistry. 57(6):922-925.
• 62. Rao, S., Sun, J., Liu, Y., Zeng, H., Su, Y., and Yang, Y., (2012). ACE Inhibitory Peptides and Antioxidant Peptides Derived from in Vitro Digestion Hydrolysate of Hen Egg White Lysozyme. Food Chemistry. 135(3):1245-1252.
• 63. Vermeirssen, V., van der Bent, A., Van Camp, J., van Amerongen, A., and Verstraete, W., (2004). A Quantitative In Silico Analysis Calculates the Angiotensin I Converting Enzyme (ACE) Inhibitory Activity in Pea and Whey Protein Digests. Biochimie. 86(3):231-239.
• 64. Escudero, E., Mora, L., and Toldrá, F., (2014). Stability of ACE Inhibitory Ham Peptides against Heat Treatment and in Vitro Digestion. Food chemistry. 161:305-311.
• 65. Świeca, M., Gawlik-Dziki, U., Dziki, D., Baraniak, B., and Czyż, J., (2013). The Influence of Protein–Flavonoid Interactions on Protein Digestibility in Vitro and the Antioxidant Quality of Breads Enriched with Onion Skin. Food Chemistry, 141(1):451-458.