Optimization of Serine Protease Purification from Mango (Mangifera indica cv. Chokanan) Peel in Polyethylene Glycol/Dextran Aqueous Two Phase System

Amid Mehrnoush,Md Zaidul Islam Sarker,Abdul Manap Mohd Yazid,Shuhaimi Mustafa

doi:10.3390/ijms13033636

Amid Mehrnoush, Md Zaidul Islam Sarker + Show 2 more

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https://doi.org/10.3390/ijms13033636

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Abstract

Mango peel is a good source of protease but remains an industrial waste. This study focuses on the optimization of polyethylene glycol (PEG)/dextran-based aqueous two-phase system (ATPS) to purify serine protease from mango peel. The activity of serine protease in different phase systems was studied and then the possible relationship between the purification variables, namely polyethylene glycol molecular weight (PEG, 4000–12,000 g·mol−1), tie line length (−3.42–35.27%), NaCl (−2.5–11.5%) and pH (4.5–10.5) on the enzymatic properties of purified enzyme was investigated. The most significant effect of PEG was on the efficiency of serine protease purification. Also, there was a significant increase in the partition coefficient with the addition of 4.5% of NaCl to the system. This could be due to the high hydrophobicity of serine protease compared to protein contaminates. The optimum conditions to achieve high partition coefficient (84.2) purification factor (14.37) and yield (97.3%) of serine protease were obtained in the presence of 8000 g·mol−1 of PEG, 17.2% of tie line length and 4.5% of NaCl at pH 7.5. The enzymatic properties of purified serine protease using PEG/dextran ATPS showed that the enzyme could be purified at a high purification factor and yield with easy scale-up and fast processing.

Highlights

Proteolytic enzyme (EC 3.4) is a class of proteins found in various sources, including animals, plants and microorganisms because of its ubiquitous nature [1]
The aim of this study is to investigate the effects of different parameters such as polyethylene glycol (PEG) molecular weight, TLL (Tie Line Length), NaCl and pH on serine protease partitioning, purification factor and yield in order to optimize the serine protease purification in PEG/dextran aqueous two-phase system (ATPS) using response surface methodology (RSM)
There was a decrease in partition coefficient at higher PEG molecular weight, because, in this condition, PEG gets more compact conformation with intermolecular hydrophobic bonds and prevents enzyme migration into the top phase, target enzymes should be transferred to the bottom phase

Summary

Introduction

Proteolytic enzyme (EC 3.4) is a class of proteins found in various sources, including animals, plants and microorganisms because of its ubiquitous nature [1]. Proteases from plants show wide substrate specificity while at the same time have a broad range of temperature, pH in the presence of additives and other organic compounds [2]. This enzyme is widely used in many industrial processes such as food, detergents, waste, pharmaceutical and leather [3]. An FAO (Food and Agriculture Organization) report states that mango production accounts for an estimated 38% of total tropical fruit output, and in the processing of this fruit, peel and kernel are two important by-products [5]. It has been discovered that there is commercial potential in mango peel as it can be a rich and cost effective source of natural enzymes [5]

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