Abstract
An Aqueous Two-Phase System (ATPS) was employed for the first time for the separation and purification of pectinase from mango (Mangifera Indica Cv. Chokanan) peel. The effects of different parameters such as molecular weight of the polymer (polyethylene glycol, 2,000–10,000), potassium phosphate composition (12–20%, w/w), system pH (6–9), and addition of different concentrations of neutral salts (0–8%, w/w) on partition behavior of pectinase were investigated. The partition coefficient of the enzyme was decreased by increasing the PEG molecular weight. Additionally, the phase composition showed a significant effect on purification factor and yield of the enzyme. Optimum conditions for purification of pectinase from mango peel were achieved in a 14% PEG 4000-14% potassium phosphate system using 3% (w/w) NaCl addition at pH 7.0. Based on this system, the purification factor of pectinase was increased to 13.2 with a high yield of (97.6%). Thus, this study proves that ATPS can be an inexpensive and effective method for partitioning of pectinase from mango peel.
Highlights
IntroductionDespite the presence of a lot of useful enzymes such as pectinase, protease, amylase and xylanase in mango peel, this potential source is currently not being commercially utilized in any way and the peel ends up being a waste product, contributing to pollution, and with the industry incurring in a high cost for its waste treatment [2]
The yield of the enzyme was increased with 4,000 g/mol Polyethylene glycol (PEG) and it seems that in this case there is suitable available free volume to accommodate pectinase in the top phase, while a decrease in pectinase yield at PEG 2,000 g/mol was observed because low molecular mass is unsuitable for sufficient partitioning of pectinase as contaminants could be transferred to the top phase with the target enzyme and as a result decrease the enzyme yield
The findings in this study showed that this purification procedure could be employed as an efficient and attractive method for the partitioning and recovery of pectinase
Summary
Despite the presence of a lot of useful enzymes such as pectinase, protease, amylase and xylanase in mango peel, this potential source is currently not being commercially utilized in any way and the peel ends up being a waste product, contributing to pollution, and with the industry incurring in a high cost for its waste treatment [2]. The so-called Aqueous Two-Phase System (ATPS), made up of two polymers or one polymer and one salt has become an attractive method in the production of industrial enzymes compared to conventional purification methods [9], Due to the fact that the scale-up of ATPS is easy, the use of this method is desirable. Separation times are short, and the technique is non-toxic [10] In such a system, manipulation of important parameters can help to achieve a desirable protein partitioning. PEG/salt ATPS was chosen to purify pectinase and important parameters affecting the partitioning and recovery of the enzyme such as molecular weight of the polymer, phase composition, pH and NaCl addition were investigated
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