Abstract
Polygalacturonases (PG) represent an important member of pectinases group of enzymes with immense industrial applications. A fungal strain Aspergillus niger MTCC478 was used for the production of polygalacturonase both under submerged and solid-state fermentation condition. Further its production was optimized under solid-state fermentation condition with media comprising of wheat bran and tea extract. Purification of an exo-PG was achieved by acetone precipitation (60–90%) and CM-cellulose column chromatography revealing 15.28-fold purification with a specific activity of 33.47 U/mg protein and 1.2% yield. A relative molecular mass of purified PG was approximately 124.0 kDa. The pH and temperature optimum was found to be 4 and 50 °C, respectively. The kcat and Km value for degradation of PGA by the purified enzyme was found to be 194 s−1 and 2.3 mg/mL, respectively. Cu2+ was found to enhance the PG activity while Ag+ completely inhibited the enzyme activity. The application of the purified PG in orange juice clarification was elucidated.
Highlights
Pectin, an acidic polysaccharide whose basic structural repeats are a-1,4-linked-D-galacturonic acid, is widely found in the middle lamella and primary cell wall of plants
A fungal strain Aspergillus niger MTCC478 was used for the production of polygalacturonase both under submerged and solid-state fermentation condition
Solid-state fermentation was found to be most effective for polygalacturonase production (Fig. 1)
Summary
An acidic polysaccharide whose basic structural repeats are a-1,4-linked-D-galacturonic acid, is widely found in the middle lamella and primary cell wall of plants. Polygalacturonases (PGs) are produced by various organisms, such as plants (Bird et al 1988; Hadfield and Benett 1998), bacteria (Jayani et al 2010; Tariq and Latif 2012; Chen et al 2014) and fungi (Martins et al 2013; Ortega et al 2014; Cheng et al 2016) Both submerged state fermentation (SmF) and solid-state fermentation (SSF) have been successfully used in pectinases production from different microbial strains (Castilho et al 2000; Silva et al 2002; Pedrolli and Carmona 2009; Dinu et al 2007). The solid-state fermentation is preferred over submerged fermentation based on the fact that it uses various agro-industrial by-products such as soy, pulps of apple, sugar beet and coffee, peels of lemon, oranges and tomato, pomace of apple and citrus fruits, sugarcane bagasse, wheat bran, etc., making the entire process cost effective (Castilho et al 2000; Yadav and Shastri 2007; Lara-Marquez et al 2011; Yadav et al 2014)
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