Inhibition Study of EDTA and PCMB on Purified Bromelain Activity from Pineapple Core [Ananas comosus (L.) Merr.] Using Ion Exchange Chromatography Column and Antiplatelet Activity Test

Nita Magfirah Ilyas, Siswati Setiasih, Sumi Hudiyono

2018

Abstract

The aim of this research was to isolate and purify bromelain from pineapple core (Ananas comosus) through fractionation using ammonium sulfate followed by dialysis and ion exchange chromatography. Fractionation of bromelain with ammonium sulfate produces highest specific activity on fraction 2 (20-50%) of 260.042 U/mg with purity level 2.548 fold compared to crude extract. Further purification by ion exchange chromatography DEAE-sepharose and CM-Sephadex C-50 showed an increase in specific activity, sequentially 500 U/mg with purity level 4.901 fold compared to crude extract and 729.167 U/mg with purity level 7.150 fold compared to crude extract. The determination of kinetics parameter of purified bromelain using LM plot gives Km and Vmax value for casein and azocasein substrate of 0.94% (w/v); 0.07 U/min and 0.87% (w/v); 0.05 U/min respectively. Bromelain fraction was inhibited competitively with EDTA and mix-inhibition type was observed in the presence of PCMB. The addition of EDTA and PCMB at a concentration of 0.5 ppm can decrease the activity of the enzyme up to 70%. In vitro study of antiplatelet agent activity using human Platelet Rich Plasma (PRP) revealed that the purified bromelain show activity as an antiplatelet agent with percentage of aggregation 20.892% and percentage of inhibition 77.994%.

Paper Citation

in Harvard Style

Ilyas N., Setiasih S. and Hudiyono S. (2018). Inhibition Study of EDTA and PCMB on Purified Bromelain Activity from Pineapple Core [Ananas comosus (L.) Merr.] Using Ion Exchange Chromatography Column and Antiplatelet Activity Test. In Proceedings of BROMO Conference - Volume 1: BROMO, ISBN 978-989-758-347-6, pages 1-1. DOI: 10.5220/0009843200002406

in Bibtex Style

@conference{bromo18,
author={Nita Magfirah Ilyas and Siswati Setiasih and Sumi Hudiyono},
title={Inhibition Study of EDTA and PCMB on Purified Bromelain Activity from Pineapple Core [Ananas comosus (L.) Merr.] Using Ion Exchange Chromatography Column and Antiplatelet Activity Test},
booktitle={Proceedings of BROMO Conference - Volume 1: BROMO,},
year={2018},
pages={1-1},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0009843200002406},
isbn={978-989-758-347-6},
}

in EndNote Style

TY - CONF

JO - Proceedings of BROMO Conference - Volume 1: BROMO,
TI - Inhibition Study of EDTA and PCMB on Purified Bromelain Activity from Pineapple Core [Ananas comosus (L.) Merr.] Using Ion Exchange Chromatography Column and Antiplatelet Activity Test
SN - 978-989-758-347-6
AU - Ilyas N.
AU - Setiasih S.
AU - Hudiyono S.
PY - 2018
SP - 1
EP - 1
DO - 10.5220/0009843200002406