An alkalophilic and thermostable polygalacturonase (PGase) from Pseudomonas sp. 13159349: purification, biochemical characterization and its efficacy in olive oil extraction

Statistical bioprocess optimization for enhanced production of a thermo alkalophilic polygalacturonase (PGase) from Pseudomonas sp. 13156349 using solid substrate fermentation (SSF)

“…The sequence was deposited in the NCBI Gene Bank with accession # OP159848.1. The enzyme secreted by this strain was further purified, and its biochemical characteristics were described in detail [ 9 ].…”

“…PGase enzyme activity was estimated using the DNS technique with citrus pectin as the substrate [ 9 ]. The experiment was carried out with 0.2 ml enzyme, 1.3 ml Tris-HCl pH 9 (optimal pH), and 0.5 ml 1% citrus pectin (Sigma-Aldrich).…”

“…Plackett-Burmann, a two-factorial experimental design was utilized to evaluate the media components affecting PGase production with wheat bran as the solid substrate. Based on bacterial growth metrics, literature review, and biochemical characteristics [ 9 ], eleven independent variables were chosen that included five physical (pH, temperature, moisture, incubation time, and inoculum size) and six nutritional (pectin, starch, (NH₄)₂SO₄, yeast extract KH 2 PO4, CoCl 2 ) factors ( Table 1 ). Design Expert STAT Ease software, version 11 (Minneapolis, USA), was used to build a random PB matrix with 13 experiments ( Table 2 ), and the impact of each variable on enzyme production was examined at both its upper (+1) and lower limit (−1) by performing the enzyme assay.…”

“…Since alkaline PGases can tolerate high pH and temperature, they are widely utilized in industrial sectors such as biofuels, production of pectin-free starch, coffee, and tea fermentation, oil extraction, bio-scouring of fabrics, polishing of jute fabrics, degumming and retting of natural fibers, paper pulp bleaching, pectic wastewater treatment, etc [ 7 , 8 ]. The application of PGase reported in this study is already established in extraction of olive oil [ 9 ]. However, the scarcity of alkaline PGase-producing strains and the optimal culture conditions prevent the large-scale production of this enzyme [ 10 , 11 ].…”