One of plant-based products for dental care is
plant-based proteolytic enzymes which are principally proteases. In order not
to damage the protein and bioactive
content, an efficient method should be employed for their purifications. As such,
three-phase partitioning (TPP) was used to purify protease from moringa (Moringa
oleifera). TPP is an emerging, promising, non-chromatographic and economical technology which is simple,
quick, efficient and often one-step process for the separation and purification
of bioactive molecules from natural sources. It involves the addition of
salt (ammonium sulphate) to the crude
extract followed by the addition of an organic solvent (butanol). The protein
appears as an interfacial precipitate between upper organic solvent and lower
aqueous phases. The various conditions such as ammonium sulphate, ratio
of crude extract to t-butanol and pH which are required for attaining efficient
purification of the protease fractions were optimized. Under optimized conditions,
it was seen that, 35% of ammonium sulphate saturation with 1:0.75 ratio of
crude extract to t-butanol at pH 7 gave 4.94-fold purification with 96.20%
activity yield of protease in the middle phase of the TPP system. The purified
enzyme from Moringa oleifera has no
antimicrobial effect on the pathogenic bacteria tested. However, this purified
enzyme, can be considered as a promising agent, cheap, and safe source which is
suitable for using in various industries.
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