“Saba”香蕉(Musa balbisiana Colla)果肉多肽抗高血压和抗氧化活性的初步评价

KIMIKA Pub Date : 2019-06-06 DOI:10.26534/kimika.v30i1.31-39
E. Magpantay, Ana Teresa B. Sucgang, M. Clemencia, T. Villar, M. Torio
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引用次数: 0

摘要

用0.125 M Tris-HCl和50 mM NaCl在pH 7.4条件下分离粗蛋白。采用Lowry法测定粗提物的蛋白质含量为4.28 mg/mL。与主蛋白对应的主带分子量约为20 KDa。分离得到的粗蛋白分别用胃蛋白酶、胰蛋白酶、凝乳胰蛋白酶和热溶酶进行酶解3、4、12和24小时。2,2-二苯基-1-吡啶酰肼(DPPH)测定结果表明,24小时消化液的抗血管紧张素转换酶(ACE)活性最高(36.02%),12小时消化液的抗氧化活性最高(33.14%)。采用超高效液相色谱法(UPLC)测定24小时消化液中负责ACE抑制活性的肽段。选取PF1、PF2和PF3三个肽段,其中PF2对ACE的抑制率最高(32.21%)。对PF2进行薄层色谱(TLC)分析,根据谷氨酸(E)(0.43)和赖氨酸(K)(0.13)的Rf值推测,PF2可能是EK。通过计算机分析,将结果与推测的具有抗氧化和抗高血压活性的肽的存在联系起来。结果表明,根据“Saba”香蕉的蛋白序列,发现了EK、GS、TY、FNE、FP、LKA、PT、PP、FAL等抗高血压肽和IR、VPW等抗氧化肽。薄层色谱法证实了抗高血压肽EK的存在。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Preliminary Assessment of the Antihypertensive and Antioxidative Activities of the Peptides from “Saba” Banana (Musa balbisiana Colla) Flesh
The crude proteins were isolated using 0.125 M Tris-HCl with 50 mM NaCl at pH 7.4. The protein content of the crude extract was determined using the Lowry assay and was found to be 4.28 mg/mL.  The major band which corresponds to the major protein has an approximate molecular weight of 20 KDa. The isolated crude proteins were subjected to enzymatic digestion using pepsin, trypsin, chymotrypsin, and thermolysin for 3, 4, 12, and 24 hours. The 24-hour digest was found to have the highest percent anti-Angiotensin Converting Enzyme (ACE) activity (36.02%) while the 12-hour digest was found to have the highest anti-oxidative activity (33.14%) using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The 24-hour digest was subjected to Ultra-Performance Liquid Chromatography (UPLC) to determine the peptide fraction responsible for the ACE inhibitory activity. Three peptide fractions (PF1, PF2 and PF3) were chosen and PF2 exhibited the highest percent inhibitory activity (32.21) against ACE. PF2 was subjected to thin layer chromatography (TLC) and the possible identity is EK based on the Rf value traveled by glutamic acid (E) (0.43) and Lysine (K) (0.13). In silico analysis was done to correlate the results with the presence of putative peptides with antioxidative and antihypertensive activities. Results showed that antihypertensive peptides EK, GS, TY, FNE, FP, LKA, PT, PP, FAL and antioxidative peptides IR and VPW were found based on the sequence of the protein in “Saba” banana. The presence of the antihypertensive peptide EK was verified using thin layer chromatography (TLC).
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