Abstract
Bromelain-generated biopeptides from stone fish protein exhibit strong inhibitory effect against ACE and can potentially serve as designer food (DF) with blood pressure lowering effect. Contextually, the DF refer to the biopeptides specifically produced to act as ACE-inhibitors other than their primary role in nutrition and can be used in the management of hypertension. However, the biopeptides are unstable under gastrointestinal tract (GIT) digestion and need to be stabilized for effective oral administration. In the present study, the stone fish biopeptides (SBs) were stabilized by their encapsulation in sodium tripolyphosphate (TPP) cross-linked chitosan nanoparticles produced by ionotropic gelation method. The nanoparticles formulation was then optimized via Box-Behnken experimental design to achieve smaller particle size (162.70 nm) and high encapsulation efficiency (75.36%) under the optimum condition of SBs:Chitosan mass ratio (0.35), homogenization speed (8000 rpm) and homogenization time (30 min). The SBs-loaded nanoparticles were characterized for morphology by transmission electron microscopy (TEM), physicochemical stability and efficacy. The nanoparticles were then lyophilized and analyzed using Fourier transform infra-red spectroscopy (FTIR) and X-ray diffraction (XRD). The results obtained indicated a sustained in vitro release and enhanced physicochemical stability of the SBs-loaded nanoparticles with smaller particle size and high encapsulation efficiency following long period of storage. Moreover, the efficacy study revealed improved inhibitory effect of the encapsulated SBs against ACE following simulated GIT digestion.
Highlights
Sea cucumber is a protein-rich marine invertebrate
The use of box behnken design (BBD) in process optimization is considered to be more effective and preferable compared to central composite design based on the less number of experiments that are involved in the former
The stone fish biopeptides (SBs) loaded Chitosan:TPP NPs were produced by ionotropic gelation method under different encapsulation conditions according to response surface methodology by Box-Behnken design (Table 1)
Summary
Sea cucumber is a protein-rich marine invertebrate. Numerous ACE-inhibitory biopeptides have been generated and characterized from different species of sea cucumber such as Stichopus horrens[6]; Actinopyga lecanora tissue proteins[7]; Stichopus vastus collagen[8]; Isostichopus badionotus[9]; Acaudina malpadioidea wall protein[10]. The bioactive peptides are susceptible to degradation by gastrointestinal enzymes and need to be protected for effective target site delivery via oral route This can be achieved through encapsulation to safe guard their structural and functional integrity and improve their stability against gastrointestinal proteases and peptidases[12]. Natural, non-toxic and biodegradable polymer made up of (β1 → 4) linked 2-amino-2-deoxy-glucopyranose (GlcN) and 2-acetamido-2-deoxy-d-glucopyranose (GlcNAc) residues[17,18] It is considered as a safe coating material for the encapsulation of bioactive compounds[19,20]. The present work was aimed to improve the physicochemical stability and efficacy of bromelain-generated ACE-inhibitory biopeptides from stone fish as edible species of sea cucumber through their encapsulation in Chitosan:TPP nanoparticles using ionotropic gelation method and optimized by Box-Behnken design
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