Isolation, purification and characterization of bioactive peptides exhibiting antihypertensive activity from freshly prepared pineapple (Ananas comosus (L) Merr.) Fruits and commercially available pineapple products

Date

4-2014

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Mary Ann O. Torio

Restrictions

Restricted: Not available to the general public. Access is available only after consultation with author/thesis adviser and only to those bound by the confidentiality agreement.

Abstract

In silico analysis was done to assess the presence of ACE-inhibiting bioactive peptides from fruit bromelain. Isolation of the crude protein (fruit bromelain) from three different varieties of fresh pineapple fruits (FMC1, FC74, FMP2) and commercially available pineapple juice (CAPJ) and chunks (CAPC) was done using an extraction buffer composed of 50 mM Tris-HCI buffer pH 7.0, 35 mM potassium phosphate buffer and 0.4 M NaCl. The crude protein was purified using ammonium sulfate (AS) precipitation, dialysis and gel filtration chromatography (GFC). The 20%-30% ammonium sulfate fractions contained the protein of interest and were further subjected to dialysis and GDC. Protein content of all samples was determined through Bradford method. Molecular weight of the crude, partially purified and purified protein was estimated through the Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoreisis (SDS-PAGE) and was approximately to be 30-35kDa. The purified protein was digested using combined pepsin, chymotrypsin and trypsin mixture (2:1:1 ratio) at 0, 3,5,10,30,60 minutes and 24 hours interval. Percent inhibition of 24-hour protein digest from fresh pineapple fruits, pineapple juice and chunks were 53.54%, 57.01%, 49.92% and 37.64% for FC74, FMP2, CAPJ and CAPC, respectively. Among all the samples, the 24-hour digest of the variety for a market (FMC1) showed the highest % ACE-inhibition of 69.84% with a corresponding IC50 value is 0.0027 mg/mL.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2014 C42 /A94

Document Type

Thesis

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