Development of a technique for obtaining bromelain associates with carboxymethyl chitosan micro- and nanoparticles
Abstract
Bromelain, a plant-derived cysteine protease, has great potential for medical and industrial applications. Soluble forms of the enzyme are subject to rapid inactivation due to conformational rearrangements caused by the influence of many chemicals and physical factors, so their use is limited. One of the ways to overcome these difficulties is to adsorb bromelain on various biopolymers. Adsorption is the simplest method of enzyme immobilisation, which influences the activity of biocatalysts. Therefore, the aim of the study was to obtain bromelain complexes with micro- and nanoparticles of carboxymethyl chitosan with and without ascorbic acid, to determine the catalytic activity of the biocatalysts, and to evaluate their stability under optimal functioning conditions.
We synthesised micro- and nanoparticles of medium molecular weight (200 kDa) and high molecular weight (350 kDa) carboxymethyl chitosans with the and without ascorbic acid, and also obtained bromelain complexes with micro- and nanoparticles of carboxymethyl chitosan. The functional activity of the preparations decreased during seven days of the experiment. The catalytic capacity of bromelain associates with carboxymethyl chitosan microparticles with molecular weights of 200 and 350 kDa increased by 63 and 52 % compared to the free enzyme. When the same complexes were prepared with ascorbic acid, the activity increased by 69 % for medium molecular weight carboxymethyl chitosan and by 55 % for high molecular weight carboxymethyl chitosan.
The catalytic capacity of bromelain associates with carboxymethyl chitosan nanoparticles with molecular masses of 200 and 350 kDa increased by 62 and 30 % compared to the free enzyme. When ascorbic acid was added, the activity of the complexes increased by 65 % for medium molecular weight carboxymethyl chitosan and by 50 % for high molecular weight carboxymethyl chitosan.
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