Purification and characterization of exo-polygalacturonase from Zygoascus hellenicus V25 and its potential application in fruit juice clarification

Abstract

The purification and characterization of the extracellular polygalacturonase from V25 submerged culture using orange peel waste were investigated. This polygalacturonase, with a molecular weight of 75.28 kDa, was purified to 16.89 purification fold with a recovery of 18.46% and specific activity of 2469.77 U/mg protein by ammonium sulfate precipitation, DEAE cellulose chromatography, and Sephadex G-100 gel filtration. The enzyme exhibited maximum activity at 60°C and pH 5.0 and was stable over a wide range of pH levels (3.0-11.0). Moreover, enzyme activity was enhanced by Cu and cysteine, whereas it was strongly inhibited by Hg. The extent of enzymatic hydrolysis was negatively correlated with the degree of pectin esterification. K and V values of the polygalacturonase were 5.44 mg/mL and 61.73 μmol/(min·mg), respectively. The polygalacturonase was applied in the juice clarification of four fruits, and results showed that the percentage transmittance at 660 nm increased by 3.51, 4.36, 8.04, and 12.2%.

A review on pectinase properties, application in juice clarification, and membranes as immobilization support

“…However, A. luchuensis and S. purpureum endoPG activity degraded in the presence of magnesium ions (Carli et al, 2019;Tan et al, 2020). ExoPG also observed the same behavior in the presence of magnesium ions (Anand et al, 2017;Chen et al, 2014;Kant et al, 2013;Lu et al, 2016;Pagnonceli et al, 2019). Cations of cadmium, copper, lead, tin, mercury, and silver showed inhibitory effects on pectinases activity (Pagnonceli et al, 2019;Poturcu et al, 2017;Saoudi et al, 2015;Tan et al, 2020).…”

“…MEY‐1 and Stereum purpureum GH28 and Aspergillus repens isolated PME expressed molecular mass of 71, 60, and 141 kDa, respectively (Arotupin et al., 2008; Carli et al., 2019; Yang et al., 2011). Whereas, ExoPG from various sources showed molecular weight between 50 and 106 kDa (Anand et al., 2017; Kant et al., 2013; Lu et al., 2016; Pagnonceli et al., 2019). The higher mass of macromolecules is attributed to glycosylation.…”

“…Thermophile isolates can be a vital tool in high‐temperature biochemical processes. Thermophiles obtained from various sources were stable at optimum temperature (Y. Chen et al., 2014; Cheng et al., 2016; Lu et al., 2016; Saoudi et al., 2015). However, S. purpureum GH28 endoPG reported 60% residual activity after 30 min of incubation at OT (Carli et al., 2019).…”

“…PME extracted from D. stramonium expressed a higher affection toward substrate with K m of 0.004 mg/ml (Dixit et al., 2013). In comparison, exoPG from Zygoascus hellenicus reported a maximum value of Michaelis’ constant, that is, 5.44 mg/ml (Lu et al., 2016). Due to nonstandardize assay conditions, parameters like pH, temperature, time, and buffer utilized for assay may vary from article to article.…”

High-yield intracellular production of an exo-polygalacturonase enzyme via heterologous expression of Penicillium notatum gene in Saccharomyces cerevisiae

“…4.2.2.9) and endolytic (Endo-PGL, E.C. 4.2.2.2) catalysis (Lu et al, 2016;Patidar et al, 2018). Depolymerases that show action by taking rhamnogalacturonan as a substrate are additionally categorised into (i) rhamnogalacturonan-hydrolysing enzyme rhamnogalacturonase (RG,E.C.3.2.1.171), and (ii) rhamnogalacturonan endolyase (RGL, E.C.…”