EXTRACTION OF COLLAGENOLYTIC ENZYME FROM FISH VISCERA BY PHASE PARTITIONING (PEG/CITRATE) AND ITS POTENTIAL FOR INDUSTRIAL APPLICATION

Jessica Costa Silva,Vagne Melo Oliveira,Ana Lúcia Figueiredo Porto,Márcia Nieves Carneiro Da Cunha,Juanize Matias Da Silva Batista,Luiz Henrique Svintiskas Lino

doi:10.20950/1678-2305.2020.46.4.593

Abstract

Internal viscera fish are potential sources of protein biomolecules of biopharmaceutical interest. However, this residue is frequently discarded inappropriately. The possibility to obtain by products of higher added value is a reality. Inside this view attention must be given to processes for the recovery and extraction of target molecules. However, the high cost of processing these residues is one of the obstacles to their reuse; techniques that facilitate their handling and make the process cheaper are desirable, such as extraction in a two-phase aqueous system. Thus, the aim of this study was to extract collagenolytic enzymes from common snook (Centropomus undecimalis) using a two-phase aqueous system (polyethylene glycol/citrate), according to the 24 factorial design, using as variables: molar mass of PEG (MPEG), PEG concentration (CPEG), citrate concentration (CCIT), pH, still, considering purification factor (FP), partition coefficient (K), and yield (Y). The collagenolytic activity of the crude extract was 102.41 U mg-1, after partitioning, was purified 3.91 times (MPEG: 8000; CPEG: 20.0%; CCIT: 20.0% and pH 6.0). Inhibition (U mg-1) was observed in benzamidine (22.51), TLCK (21.05), TPCK (21.29), and PMSF (23.05), signaling to be a serine-protease. The results showed the advantage of this semipurification technique as concerns to the low cost of extraction and purification, adding value to the fishing source material and allocating the residues from its processing to the industry.

Highlights

MATERIAL AND METHODSThe need for reuse of waste is a practice that has been increasingly encouraged, especially organic solids, such as those from the fishing industry
The results showed the advantage of this semipurification technique as concerns to the low cost of extraction and purification, adding value to the fishing source material and allocating the residues from its processing to the industry
The effect of independent variables and their interactions evaluated of factorial design showed that K was positively influenced by the polyethylene glycol (PEG) molar mass (MPEG) (Table 3)

Summary

Introduction

MATERIAL AND METHODSThe need for reuse of waste is a practice that has been increasingly encouraged, especially organic solids, such as those from the fishing industry. From digestive viscera of aquatic organisms, it is possible to extract various proteases (pepsin, trypsin, chymotrypsin and collagenases and/or enzymes with similar catalytic functions) (Oliveira et al, 2017a; Gurumallesh et al, 2019; Silva et al, 2019). Enzymes with collagenolytic properties of aquatic organisms have already been recovered and partially purified from digestive residues of Neotropical fish species (Silva et al, 2019; Oliveira et al, 2017b; Liu et al, 2019). The collagenolytic property (collagen cleavage) is quite required in industry, mainly biomedical and pharmaceutical, being applied in surgical procedures (Zhao et al, 2019), intracerebral therapies (Chen-Roetling et al, 2019), wound healing (Abood et al, 2018), extraction of collagen and production of bioactive peptides (Bhagwat and Dandge, 2018), fibrosis treatment (Villegas et al, 2018), and treatment of Dupuytren (Zhang et al, 2019), making these enzymes fall into the category of high commercial value proteases (Oliveira et al, 2017c)

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Conclusion