PARTIAL PURIFICATION OF MILK-CLOTTING ENZYME FROM THE SEEDS OF MORINGA OLEIFERA

Amna E Tajalsir,Fatima J Khider,Ali S A Ebraheem,Abdelmoez M Abdallah,Isam A Mohamed Ahmed,Mohamed O Elsamani

doi:10.15414/jmbfs.2014.4.1.58-62

Amna E Tajalsir, Fatima J Khider + Show 4 more

Open Access

https://doi.org/10.15414/jmbfs.2014.4.1.58-62

Copy DOI

Abstract

The aim of the present study was to search for milk clotting substitute from different parts (flowers, seeds, stem, leaves, ripe and unripe fruits) of Moringa oleifera. The samples were blended and extracted using different types of extracting solutions. The most reliable, quick and efficient enzyme extracting solution was found to be 5% NaCl in 100 mM sodium acetate buffer, pH 5.0, which was used throughout the study. The milk clotting activity was only observed in the seeds extract while the other parts were either deficient or has very low milk clotting activity. Thus, the moringa seeds were used as source of milk clotting enzyme. The extracted proteins were fractionated with ammonium sulfate at concentration of 20, 30, 40, 50 and 60 %. Highest milk clotting activity was observed in the 20 % fraction. This fraction was assumed to contain the clotting enzymes and characterized for its heating stability (30 – 90°C) and optimum temperature (30 – 90°C). The results demonstrated that moringa seeds milk clotting enzyme is stable up to 50°C with an optimum milk clotting activity of 70°C. The high ratio of milk-clotting to proteolytic activity of the partially purified enzyme indicates the potential of this enzyme as suitable rennet substitute in dairy industry. However, further study is needed to completely purify and characterize this promising milk clotting enzyme from moringa seeds.

Highlights

Cheese is the generic name for a group of fermented milk-based food products, with more than 1000 types, produced in a wide range of flavours and forms throughout the world that has played a key role in human nutrition for more than 8000 years (Fox & McSweeney 2004)
All these reasons have necessitated the search for new protease with high specific milk-clotting activity and low general proteolytic activity to be used as a rennet substitute
Microbial rennets produced by genetically engineered bacteria have proven to be suitable substitutes for animal rennet, but increasing attention has been directed toward natural rennet extracted from plants such as Ananas comosus, Carica papaya, Calotropis porcera, Ficus carica, Calm viscera, Cynara cardunculus (Roseiro et al, 2003), Cynara scolymus (Sidrach et al, 2005), and Solanum dubium (Mohamed Ahmed et al, 2009 a, b) among others

Summary

Introduction

Cheese is the generic name for a group of fermented milk-based food products, with more than 1000 types, produced in a wide range of flavours and forms throughout the world that has played a key role in human nutrition for more than 8000 years (Fox & McSweeney 2004). Microbial rennets produced by genetically engineered bacteria have proven to be suitable substitutes for animal rennet, but increasing attention has been directed toward natural rennet extracted from plants such as Ananas comosus, Carica papaya, Calotropis porcera, Ficus carica, Calm viscera, Cynara cardunculus (Roseiro et al, 2003), Cynara scolymus (Sidrach et al, 2005), and Solanum dubium (Mohamed Ahmed et al, 2009 a, b) among others Most of these plant rennets were found inappropriate because they possess high general proteolytic activity, which leads to the production of short peptides that are responsible defect in flavor and texture of cheese (Anusha et al, 2014). This enzyme was further partially purified by using ammonium sulfate precipitation techniques, and milkclotting properties and temperature profile of the partially purified enzyme were evaluated

Objectives

Results

Conclusion