Isolation and purification and characterization of the major storage protein from winged bean (psophocarpus tetragonolobus (L.) DC) and characterization of its anti- hypertensive property

Date

10-2010

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Mary Ann O Torio

Co-adviser

Mark Rickard N. Angelia

Committee Member

Roberta N. Garcia

Abstract

Total soluble proteins of winged bean var. Gloria, was extracted using a buffer containing 0.1 M Tris-HC1, pH 8.3, and 35 Mm phosphate buffer and 0.4 M NaCl. Crude protein concentration was determined using Bradford Assay. The amount of crude protein was purified using ammonium sulfate fractionation and hydrophobic interaction chromatography. SOS — PAGE Profile showed that the purified protein has two bands corresponding to 19333 Da and 29000 Da. The purified protein was subjected to protein digestion using pepsin. Results showed that the purified protein was completely hydrolyzed after 24 hrs. digestion time. The peptic hydrolysate was further subjected to Angiotensin Converting Enzyme (ACE) inhibitory activity. The ACE inhibitory activity of the peptic hydrolysate was 9.38 x 10 U and showed percent inhibition of 37.06%. Reverse Phase High Performance Liquid Chromatography (HPLC) was done to peptic hydrolysates to further separate and purify the bioactive peptides. Three randomly selected (with highest peaks) fractions were again assayed for ACE inhibitory activity. Results showed that the inhibitory activities obtained were 1.4492 a 10.3U for fraction 4, 1.692 a 10 U for fraction 8, and 4.872 x 10 -3 U for fraction 9. The percent inhibition of the three fractions were 57.965, 64.38%, and 80.75%, respectively. Fraction 9 has the highest inhibitory activity and percent inhibition and this was further subjected to Thin Layer Chromatography. From the results, the possible sequence of the bioactive peptide fragment was VK for it has the closest values to the standard amino acid Rf values.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2010 C4 M33

Document Type

Thesis

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