Extraction and crosslinking of bromelain aggregates for improved stability and reusability from pineapple processing waste

The present study is first of its kind that focuses upon the extraction of bromelain from pineapple core waste and stabilising it as insoluble cross-linked aggregates. The influence of process variables such as the choice of precipitant, type of cross-linker, concentration of cross-linker and the reaction time for cross-linking step was investigated upon the activity recovery of bromelain cross-linked aggregates. The optimization of this biocatalyst preparation specifically recovered 87% of the enzymatic activity available in pineapple core waste by ammonium sulphate (60%, w/v) precipitation followed by cross-linking for 4 h with 80 mM glutaraldehyde. Cross-linked bromelain aggregates were thermally more stable and exhibited higher pH stability in comparison to free bromelain. The cross-linked bromelain aggregates exhibited higher operational stability in different organic solvents at 4 °C. The highest operational stability (% stability given in parenthesis) was observed in acetone (100%) followed by hexane (53.6%), ethyl acetate (39.6%), ethanol (32.5%) and chloroform (14.9%). The kinetic studies revealed higher Km value (5.45 mM) after the formation of cross-linked bromelain aggregates as compared to free bromelain (5.04 mM) with almost similar Vmax values. Cross-linked bromelain aggregates also showed significant reusability characteristics with an activity retention of >85% after 5-time cycles. Such recyclability of bromelain cross-linked aggregates could lead to potential industrial applications in both food and non-food sector. In addition, the present extraction method avoids costs related to purification and expensive immobilization carriers.