Bromelain from pineapple stem in alcoholic–acidic buffers for wine application

Abstract

a b s t r a c tBromelain from pineapple stem has been studied in unexplored model wine buffer (pH 3.2 tartaric acid and ethanol) in order to evaluate its applicability for white wine protein stabilization.Bromelain proteolytic activity was studied against several synthetic peptide substrates in McIlvaine buffer covering a wide pH range. Among different synthetic substrates, Bz-Phe-Val-Arg-pNA turned out to be the most suitable one to test bromelain activity at the average minimum pH value of wine (3.2). The protease activity was significantly increased by the presence of 5 mM cysteine while no influence was exerted by EDTA. The inhibiting effect of ethanol was investigated in the range 0-25% (v/v), resulting in being rather limited at the average minimum concentration found in wine (10% v/v).The kinetic parameters were estimated in both McIlvaine (as reference) and model wine buffers. This study showed that stem bromelain worked well at the average minimum pH value of wine, and had a significantly higher activity in model wine buffer (K a increased of 42%). These results are the basis for suggesting a future biotechnological application of this protease in winemaking.

Kinetic studies on recombinant stem bromelain

“…In addition, both enzymes had broad specificities, with ZPVANA being most susceptible and ZANPE being the least susceptible. The optimum pH of 8 observed for ZPVANA and ZAANA substrates is similar to that reported by Benucci, et al [22] on stem bromelain. However, the authors obtained higher specific activities with ZAANA instead of ZPVANA.…”

Stability, purification, and applications of bromelain: A review

“…Bromelain is also useful in industrial biotechnology, especially in winemaking, since it prevents the formation of unwanted protein precipitates after the wine is bottled. 69 Esti et al 70 studied the effect of wine inhibitors (ethanol, sulfur dioxide, grape skin, and seed tannins) on the activity of stem bromelain covalently immobilized on chitosan beads. The results obtained showed that immobilized stem bromelain might have potential biotechnological applications in winemaking, since this enzyme is capable of stabilizing proteins in wine.…”

Degradation of white wine haze proteins by Aspergillopepsin I and II during juice flash pasteurization

“…For these reasons, alternative methods for white wine stabilization have been extensively investigated. A variety of alternatives has been proposed, ranging from the use of other adsorbents (Cabello-Pasini, Victoria-Cota, Macias-Carranza, Hernandez-Garibay, & Muñiz- Salazar, 2005;de Bruijn et al, 2009;Salazar, Achaerandio, Labbe, Guell, & Lopez, 2006;Sarmento, Oliveira, & Boulton, 2000;Vincenzi, Polesani, & Curioni, 2005), ultrafiltration (Hsu et al, 1987), flash pasteurization (Pocock, Høj, Adams, Kwiatkowski, & Waters, 2003) and proteases (Benucci, Liburdi, Garzillo, & Esti, 2011;Waters, Wallace, & Williams, 1992), but none has proven sufficiently effective to replace bentonite.…”