John Leanard M. Lasquite
◽
Brendaline C. Enopia
◽
Roger Francis E. Senangote
◽
Danilo C. Sabarre Jr.
◽
Camila Flor Yagonia-Lobarbio
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 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 (K E ) 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.
A novel and environmentally friendly bioprocess for separation and partial purification of polysaccharides from Cordyceps sinensis mycelia by an aqueous two-phase system