Partial Purification of Polyphenol Oxidase (PPO) from Mango Seed Kernel (MSK) Using Alcohol/Salt-Based Aqueous Two-Phase System (ATPS)

Title:Partial Purification of Polyphenol Oxidase (PPO) from Mango Seed Kernel (MSK) Using Alcohol/Salt-Based Aqueous Two-Phase System (ATPS)

Volume: 10 Issue: 2

Author(s): John Leanard M. Lasquite, Brendaline C. Enopia, Roger Francis E. Senangote, Danilo C. Sabarre, Jr. and Camila Flor J. Yagonia-Lobarbio*

Affiliation:

• BioProcess Engineering and Research Center, Department of Chemical Engineering, University of San Carlos, Talamban, Cebu City 6000, Philippines

Keywords: Polyphenol oxidase, ATPS, partial purification, mango seed kernel, crude enzyme extract, PPO.

Abstract: Background: Polyphenol oxidase (PPO) has applications in pharmaceutical, environmental, and food industries. However, conventional techniques for its purification are considered costly and laborious. Alternative methods of purification are needed.

Methods: This study explored the use of an alcohol/salt-based Aqueous Two-Phase System (ATPS) to partially purify PPO from Mango Seed Kernel (MSK). Specifically, this study determined the effects of the 1-propanol/potassium phosphate buffer mass ratio on the partition parameters such as partition coefficients, activity recovery, and purification fold.

Results: The enzyme partition coefficient (KE) was greater than 1, which signified that more PPO was partitioned in the top phase due to its hydrophobic nature. Specifically, the 3.22 1-propanol/ PPB mass ratio had the highest K E of 1.91 ± 0.16 and 1.46 ± 0.16 for runs 1 and 2, respectively. Moreover, the same mass ratio led to the highest activity recovery in the top phase reaching up to 73.99 ± 1.56% for run 1 and 64.27 ± 2.48% for run 2. In terms of purification fold, runs 1 and 2 had PF values of 0.55 ± 0.05 and 0.70 ± 0.01, respectively. Since the values obtained were low due to the large number of contaminants extracted together with the target enzyme in the top phase, the use of a back-extraction step recovered the PPO in the bottom phase and resulted in an activity recovery of 82.94 ± 1.05% and a purification fold of 7.01 ± 0.95.

Conclusion: Results have shown that varying the mass ratio had significant effects on the partition parameters.

Cite this article as:

Lasquite Leanard M. John, Enopia C. Brendaline, Senangote Francis E. Roger , Sabarre, Jr. C. Danilo and Yagonia-Lobarbio Flor J. Camila*, Partial Purification of Polyphenol Oxidase (PPO) from Mango Seed Kernel (MSK) Using Alcohol/Salt-Based Aqueous Two-Phase System (ATPS), Current Biotechnology 2021; 10 (2) . https://dx.doi.org/10.2174/2211550110666210603105630