Kinetic Studies of Purified Bromelain from Pineapple (Ananas comosus [L.])Merr) Core with Hidroxyapatite and CM Sephadex C-50 Ion Exchange Chromatography

Nofa Rahayu Desi Putri, Sumi Hudiyono, Siswati Setiasih

2018

Abstract

Bromelain is a protease enzyme that functions to break peptide bonds. in the health field bromelain can function as anti-inflammatory, anti-thrombotic and fibrinolytic. Here we purposed a new purification step to collect enzyme from pineaplle. The purification of the enzyme from Ananas comosus was carried out by precipitation with varying concentration of acetone and followed by column chromatography using hidroxyapatite ion exchage and CM sephadex C-50 resin. The enzyme activities were evaluated using casein as substrate. The highest specific activity of bromelain was gained from acetone fractination as 51,51 U/mg in the range of 50-80% saturation with purity level of enzyme as 11 times from its crude extract. The highest specific activity from last fraction which run on hidroxyapatite ion-exchange column chromatography was 87,49 units/mg with purity level of enzyme resulted 19 times higher compared to the crude extract. Later on, the purification with CM sephadex C-50 resulted in increasing the specific activity to 200 U/mg with purity level of enzyme as 45 times higher from its crude extract. Hydrolysis of various casein concentration with purified bromelain was carried out at optimum reaction condition of pH 7,0 and 37 0 C. The results obtained revealed the Km and Vmax value were 0,94% (w/v) and 0,023 U/min respectively. From the various purification steps that have been done, it can be observed the increasing of bromelain specific activity from each stage.

Paper Citation

in Harvard Style

Putri N., Hudiyono S. and Setiasih S. (2018). Kinetic Studies of Purified Bromelain from Pineapple (Ananas comosus [L.])Merr) Core with Hidroxyapatite and CM Sephadex C-50 Ion Exchange Chromatography. In Proceedings of BROMO Conference - Volume 1: BROMO, ISBN 978-989-758-347-6, pages 1-1. DOI: 10.5220/0009843300002406

in Bibtex Style

@conference{bromo18,
author={Nofa Rahayu Desi Putri and Sumi Hudiyono and Siswati Setiasih},
title={Kinetic Studies of Purified Bromelain from Pineapple (Ananas comosus [L.])Merr) Core with Hidroxyapatite and CM Sephadex C-50 Ion Exchange Chromatography},
booktitle={Proceedings of BROMO Conference - Volume 1: BROMO,},
year={2018},
pages={1-1},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0009843300002406},
isbn={978-989-758-347-6},
}

in EndNote Style

TY - CONF

JO - Proceedings of BROMO Conference - Volume 1: BROMO,
TI - Kinetic Studies of Purified Bromelain from Pineapple (Ananas comosus [L.])Merr) Core with Hidroxyapatite and CM Sephadex C-50 Ion Exchange Chromatography
SN - 978-989-758-347-6
AU - Putri N.
AU - Hudiyono S.
AU - Setiasih S.
PY - 2018
SP - 1
EP - 1
DO - 10.5220/0009843300002406