Preparation of Cicer Artienium Amylase Loaded BSA Nanoparticles and Their Bioproteolysis to be Used as Detergent Additive

Kirti Rani,Chanchal Chauhan

doi:10.13189/bb.2015.030501

Abstract

Cicer aritenium is rich source of amylase and used for extraction of amylase. Amylases are known for their industrial application in food, pharmaceutical, detergent, textile and leather industries. The extracted amylase was encapsulated into biochemically engineered BSA nanoparticles with butanol through glutaraldehyde by desolavtion method. BSA was chosen as a biocompatible matrix used for the binding of amylase where 77.6% of enzyme encapsulation was achieved. This binding was lead to enhancing the thermal stability at 72℃ for 6 hours and storage stability at 4℃ of bound enzyme for 16 months as compared to free enzyme whose thermal stability at 72℃ only for 15 minutes and storage stability only for 24 hours day were found. However, studied thermal stability, storage stability and % of encapsulation for maximal activity for encapsulated enzyme was found to be at remarkably higher as compared to free enzyme which increased the industrial viability of amylase after immobilization. Characterization of prepared nanoparticles was done by Dynamic Light scattering (DLS) and Scattering Light Microscopy (SEM). Observed size of enzyme loaded nanoparticles was observed in the range of 56 nm to 107.4 nm. Bioproteolysis of enzyme loaded BSA nanoparticles was performed by alkaline protease (10 U, 15 U, 20 U, 25 U, 30 U, 35 U, 40 U, 45 U) for sustained and controlled release of encapsulated amylase from prepared nanoparticles. And, further, their application was studied with detergents powder (Aerial, Surf Excel, Wheel and Tide) for washing of tough stained cloths pieces having chosen stains of coffee, tea, pomegranate and turmeric. These strains are found to be difficult to vanish in one simple wash. But, this cost effective bio-active enzyme loaded BSA nanopreaparation were found to be effective in washing with chosen detergents for quick and efficient washing with absolute clear visible results as compared to the results when washing of stained cloths pieces was done with detergents only.

Highlights

Amylases are starch-degrading enzymes that require calcium ions (Ca2+) for their activity, structural integrity and stability.[1]
Cicer aritenium amylase loaded biochemically engineered BSA nanoparticles has 77.6% retention of enzyme activity whose result was found to be similar to previous study in which Cicer arietinum amylase had 81.34 % retention activity of enzyme into emulsified bovine serum albumin nanoparticles[54] and lesser than 99% of Glucose oxidase was immobilized in bovine serum albumin nanoparticle using mustard oil.[46]
Cicer arietinum amylase was encapsulated into biochemically activated BSA nanoparticles by desolvation method with 77.6% of enzyme encapsulation

Summary

Introduction

Amylases are starch-degrading enzymes that require calcium ions (Ca2+) for their activity, structural integrity and stability.[1]. DEAE-cellulose was chosen for enzyme entrapment within alginate beads with 88.9% of enzyme retention after immobilization and within hydrophilic silica gel with 92.3% immobilization.[21] Other nanotechnological techniques like desolvation, emulsification, thermal gelation, nano-spray drying, nab-technology and self-assembly that have been investigated for fabrication of albumin nanoparticles.[22] Among the potential methods, emulsion formation, desolvation, or coacervation methods are commonly preferred for nanopreparation. Their bioproteolysis application was study for efficacy in washing of the stained clothes with chosen detergents powder (Aerial, Surf Excel, Wheel and Tide) for washing of tough stained cloths pieces having routine stains of coffee, tea, pomegranate and turmeric.[35,40]

Materials and Methods

Result and Discussion

Findings

Conclusions