Sulfanilic acid‐modified chitosan mini‐spheres and their application for lysozyme purification from egg white

Abstract

A cation exchange matrix with zwitterionic and multimodal properties was synthesized by a simple reaction sequence coupling sulfanilic acid to a chitosan based support. The novel chromatographic matrix was physico-chemically characterized by ss-NMR and ζ potential, and its chromatographic performance was evaluated for lysozyme purification from diluted egg white. The maximum adsorption capacity, calculated according to Langmuir adsorption isotherm, was 50.07 ± 1.47 mg g while the dissociation constant was 0.074 ± 0.012 mg mL . The process for lysozyme purification from egg white was optimized, with 81.9% yield and a purity degree of 86.5%, according to RP-HPLC analysis. This work shows novel possible applications of chitosan based materials. The simple synthesis reactions combined with the simple mode of use of the chitosan matrix represents a novel method to purify proteins from raw starting materials. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:387-396, 2018.

Lactoferrin purification and whey protein isolate recovery from cheese whey using chitosan mini-spheres

“…Chitosan mini-spheres (1.32 ± 0.14 mm diameter) were obtained according to the procedure described in previous studies (Baieli et al, 2014b;Hirsch et al, 2018a). Briefly, a chitosan solution was prepared by dissolving 2% of low viscosity chitosan powder (acetylation degree 41%) in 2% acetic acid (Kasaai, Arul, & Charlet, 2000).…”

“…The ζ potential study was performed as previously reported (Hirsch et al, 2018a). Briefly, the matrices were first disrupted mechanically (100 mg 10 mL -1 distilled water) with a spatula and then kept under magnetic stirring for 48 h to obtain a homogeneous microparticle suspension.…”

“…2. The 13 C CP-MAS spectrum for the GTMAmodified chitosan mini-spheres shows the carbon resonance signals for the chitosan structure (C 1-8 ) and those for the crosslinking with epichlorohydrin (C 9-10 ) (Hirsch et al, 2018a) and with the reaction with GTMA (C 11-14 ). Particularly, the C CPPI experiment allowed us to unequivocally assign the methyl carbons of the trimethylammonium group of the GTMA at a carbon chemical shift of 54.9 ppm (Supplementary material Fig.…”

“…An important difference between triazine dyes and their precursor is that the latter does not have regulatory restrictions as a ligand if the purified product or the matrix itself is applied to the food industry. The multimodal matrix developed was successfully tested for lysozyme purification from egg white in a previous study (Hirsch et al, 2018a). Given that LF has a high pI and a complex source, in this study the purification process parameters were optimised through statistical analysis for its purification from cheese whey using the sulfanilic acid-modified chitosan mini-spheres.…”

Enhanced chitin gel with magnetic nanofiller for lysozyme purification

“…Hydrogels based on bio-composites polymers have been intensively studied for protein purification [5]. Compared with the widely studied hydrogels based J o u r n a l P r e -p r o o f on synthetic polymers, such as acrylic acid, acrylamide and poly(ethylene glycol), the hydrogels based on biopolymers, such as alginate [6], carboxymethyl cellulose [7], silk [8], chitosan [9]and chitin derived polymers [10], have recently captured researchers interest due to their biocompatibility, nontoxicity and biodegradability.…”