Concentration by membrane separation processes of a medicinal product obtained from pineapple pulp

Abstract

The concentration of pineapple juice is needed to retain the bromelain activity and to standardize the composition and proteolytic activity. Thus, this work aimed to obtain a pure bromelain extract from the Ananas comosus L. Merril juice by membrane separation process. A 2 2 experimental planning was used to study the influence of pH and transmembrane pressure on the activity recovery by micro-filtration using a plain membrane. In second step, this enzyme was purified by the ultra-filtration using a 10 kDa millipore kit. The best operation condition to bromelain concentration using the plain membrane was at pH 7.5 and transmembrane pressure of 0.05 bar, while 85% of bromelain activity was recovered. Ultra-filtration retained 100% of proteolytic activity and concentrated in 10 fold the bromelain extract. SDS-PAGE electrophoresis showed that the ultra-filtrated had high purity and the bromelain from A. comosus pulp had a molecular weight of 24.5 kDa.

Bromelain: Methods of Extraction, Purification and Therapeutic Applications

“…While the optimum pH range for FBM is 3-8 and optimum temperature ranges from 37-70 ºC (Lopes et al 2009;Jutamongkon and Charoenrein 2010;Silvestre et al 2012). Likewise, the molecular weight range for SBM is 26-37kDa (Grzonka et al 2007;Kumar et al 2011) and for FBM molecular weight range is 24.5-32kDa (Grzonka et al 2007;Lopes et al 2009;Gautam et al 2010). When bromelain is kept at -20 ºC its stability remains intact for a defined time period (Rowan et al 1990).…”

“…The reported optimum pH range for SBM by various researchers is 6-7 and its optimum temperature range is 50-60 ºC (Harrachi et al 1998;Gautam et al 2010;Xue et al 2010). While the optimum pH range for FBM is 3-8 and optimum temperature ranges from 37-70 ºC (Lopes et al 2009;Jutamongkon and Charoenrein 2010;Silvestre et al 2012). Likewise, the molecular weight range for SBM is 26-37kDa (Grzonka et al 2007;Kumar et al 2011) and for FBM molecular weight range is 24.5-32kDa (Grzonka et al 2007;Lopes et al 2009;Gautam et al 2010).…”

“…Membrane filtration is being used for the isolation and purification of bromelain. Simultaneous use of microfiltration and ultrafiltration in order to separate bromelain from pineapple pulp resulted in active recovery of 85% (through microfiltration) and 10 fold concentrated bromelain (through ultrafiltration) (Lopes et al 2009). When microfiltration, ultrafiltration, ammonium sulfate precipitation and ultracentrifugation were employed in a sequence 98% yield was achieved (Doka et al 2005).…”

“…Efforts are being made by researchers to achieve highly purified bromelain in less steps and low cost. Modern strategies, such as membrane filtration (Lopes et al 2009), reverse micellar systems (Upadhyay et al 2010;Kumar et al 2011), aqueous two phase extraction (Coelho et al 2013;Novaeset al 2013) and chromatographic techniques (Yin et al 2011) have shown promise in this regard. Pineapple plant also contains minor quantities of other proteinases like ananain and comosain (Nadzirah et al 2013) but bromelain is regarded as a primary and extensively investigated component (Amini et al 2016).…”

The effect of pH on bromelain partition from Ananas comosus by PEG4000/phosphate ATPS

“…A study of bromelain partitioning in twophase aqueous systems containing PEO-PPO-PEO block copolymers was made an value 1.25 was the maximum purification factor found for the bromelain recovery from pineapple fruit via twophase aqueous extraction (Rabelo et al, 2004). Lopes et al (2009) purified a bromelain from pineapple juice in a continuous system of the membrane separation process. The best operation condition to bromelain concentration using the plain membrane was at pH 7.5 and transmembrane pressure of 0.05 bar, while 85% of bromelain activity was recovered.…”