Performance of a Two-Stage Membrane System for Bromelain Separation from Pineapple Waste Mixture as Impacted by Enzymatic Pre-Treatment and Diafiltration

Abstract

SummaryMembrane technology has been successfully applied for the purification of bromelain, a protease enzyme from pineapple. However, the current system operates less optimally in terms of flux and separation primarily due to properties of the feed, such as viscosity. Hence, in this study, enzymatic pre-treatment and diafiltration operation were employed in a two-stage ultrafiltration (UF) system to enhance the performance of the purification and concentration process of bromelain enzyme from an extract of pineapple crude waste mixture (CWM). Pre-treatment of the CWM extract using either pectinase or cellulase, or the combination of both, was applied and compared regarding the apparent viscosity reduction. Diafiltration step was introduced in UF stage 2 and observation on the flux performance, enzyme recovery and enzyme purity were performed. A 12 % apparent ftb.com.hr Please note that this is an unedited version of the manuscript that has been accepted for publication. This version will undergo copyediting and typesetting before its final form for publication. We are providing this version as a service to our readers. The published version will differ from this one as a result of linguistic and technical corrections and layout editing.Food Technology and Biotechnology 56 (2) 2018 www.2 viscosity reduction has been achieved when the CWM extract was pre-treated with pectinase which led to 37-38 % improvement in the flux performance of both UF stages, as well as higher enzyme recovery in UF stage 1. The introduction of diafiltration mode in UF stage 2 has managed to sustain high flux values while yielded 4.4-fold enzyme purity (higher than a 2.5-fold purity achieved in our previous work); however, high diluent consumption was needed. The outcomes of this study showed that the flux performance and bromelain separation can be enhanced by reducing the viscosity with the employment of enzymatic pre-treatment and diafiltration operation. Thus, both techniques can be potentially applied in a large-scale membrane-based process for bromelain production.

Fractionation and activity profiling of fruit bromelain from pineapples of Phuket variety growing in Thailand

“…The purified bromelain has a high level of purity and only one band was observed on SDS‐PAGE (Figure (c)). Its molecular weight has been found to be 26.5 kDa, which is consistent with other research on fruit bromelain (Bresolin et al, ; Ketnawa et al, ; Nor et al, ). The pH profiles of fruit bromelain extracted from different pineapple varieties are somehow different.…”

“…This method was characterized by low purity and high salt concentration, therefore, its application is quite limited. So far, plenty of new methods have been reported, for example, reverse micellar extraction (Umesh Hebbar, Sumana, & Raghavarao, ), block copolymers partition (Rabelo, Tambourgi, & Pessoa, ), and membrane separation (Nor, Ramchandran, Duke, & Vasiljevic, ). These methods are not easy to apply during the manufacturing process as they require certain skills.…”

Latest Advances in Protein-Recovery Technologies from Agricultural Waste

“…This is due to the removal of lower-molecular-weight compounds from the retentate, which could affect amylase activity [41]. The efficiency of a twostage membrane technique for purifying protease from a pineapple waste mixture was evaluated [42]. Two-stage ultrafiltration, enzymatic pretreatment, and diafiltration were included in this system.…”

Performance of Single and Two-Stage Cross-Flow Ultrafiltration Membrane in Fractionation of Peptide from Microalgae Protein Hydrolysate (Nannochloropsis gaditana)

“…Meanwhile, for two-stage membrane 10/5 kDa, the total permeate flux was higher than the single membrane. One plausible reason for this could be due to the change in the MPH's viscosity from using a two-stage membrane [24]. During the first stage of filtration using the 10 kDa membrane, most macromolecules larger than 10 kDa were retained in the feed.…”

“…Roslan et al [22] added that the implementation of UF membrane to the separation of protein hydrolysate is indeed a complex process. Fractionation using a two-stage UF membrane system has been studied by several researchers and has been found to improve the permeate flux and selectivity of targeted bioactive compounds [23,24]. However, there is currently a lack of studies in the literature dealing with the separation of multi-components of microalgal biomass [25].…”