Separation and Utilization of Pectin Lyase from Commercial Pectic Enzyme via Highly Methoxylated Cross-Linked Alcohol-Insoluble Solid Chromatography for Wine Methanol Reduction

Abstract

The isolation and utilization of pectin lyase (PL) from commercial pectic enzyme for methanol reduction in wine production was investigated. PL can be separated from pectinesterase (PE) and polygalacturonase (PG) on HM-CL-AIS affinity chromatography at pH 4; however, it is difficult to further distinguish PE from PG. Some desirable physicochemical properties such as transmittance, lightness, redness, and lower total pectin content are found in the external enzyme adding groups (PL, PE and PG, and pectic enzyme groups) in comparison to the control group. Methanol contents in pectic enzyme and the PE and PG groups increase from 628 +/- 13 (control group) to 3103 +/- 16 and 1736 +/- 67 mg/L ethanol in the final products, respectively. Nevertheless, the adding of PL does not cause any increase in methanol content. The results present in this study suggest that the HM-CL-AIS column is a simple, inexpensive, convenient, and effective method for PL purification. Moreover, the partial purified PL is a potential replacement of commercial pectic enzyme for pectin depolymerizing, methanol content reducing, and wine quality improving in wine production.

Effect of storage conditions on methanol content of fruit and vegetable juices

“…However, levels above the given limits were reported by some authors (Wu et al, 2005(Wu et al, , 2007. Recently, more and more people attend to drink juices from fruit and vegetables for nutrition balance and health improving in Taiwan.…”

“…PG activity was determined by measuring the release of reducing groups from 0.3% citrus pectin (DE 60-66%) in 0.2 M acetate buffer, pH 4 using a 3,5-dinitrosalicylic acid (DNS) reagent assay and monitored the absorbance of resulting colored mixture at 540 nm according to Wu et al (2007). One unit of PG activity is defined as the amount of enzyme catalyzed the liberation of 1 mmol of galacturonic acid per min at 37 1C.…”

“…Several studies have found that methanol can be produced from the hydrolysis of methyl ester groups in pectins by PE and occurs naturally at a low level in fresh fruit juices and most of the alcoholic beverages. The addition of pectolytic enzymes for ciders (Massiot et al, 1994) or wines (Servili et al, 1992;Frenkel et al, 1998;Revilla and Gonzá lezSanJosé , 1998;Cabaroglu, 2005;Wu et al, 2005Wu et al, , 2007 caused an increase in methanol level during the mashing, fermentation and aging stages.…”

“…Methanol content of juice will increase as a consequence of enzymatic degradation of pectins by endogenous PE after squeezing, and cause a risk for consumers. What concerns researchers is the level of methanol intake from wine and spirits (Servili et al, 1992;Frenkel et al, 1998;Revilla and Gonzá lez-SanJosé , 1998;Cabaroglu, 2005;Wu et al, 2005Wu et al, , 2007Zocca et al, 2007).…”

Biotechnological potential of pectinolytic complexes of fungi

“…The use of pectinolytic enzymes is being implemented in various production processes, mainly in the food industry. In juice and wine production, enzymes have been used to improve the yield, decrease the viscosity, clarify the juices and make them more stable (Kashyap et al 2001;Wu et al 2007). Unicellular products used for the preparation of nectars, baby food and yoghurt are generated through selective hydrolysis of the polysaccharides of the middle lamella to preserve the integrity of the cell wall (Kashyap et al 2001).…”