Bovine lactoferrin purification from whey using Yellow HE-4R as the chromatographic affinity ligand

Abstract

The worldwide production of whey increases by around 186 million tons each year and it is generally considered as a waste, even when several whey proteins have important economic relevance. For its valorization, inexpensive ligands and integrated chromatography methods need to be developed for specific and low-cost protein purification. Here, we describe a novel affinity process with the dye Yellow HE-4R immobilized on Sepharose for bovine lactoferrin purification. This approach based on a low-cost ligand showed an efficient performance for the recovery and purification of bovine lactoferrin directly from whey, with a yield of 71% and a purification factor of 61.

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

“…In a previous work, Fee and Chand established that there was a given value for the relation between the sample volume and the matrix amount, in which by increasing the matrix quantity there was no increase in the target protein bound to it, but there was a reduction of the time required to complete the adsorptive step. This effect was also verified in a previous work published by Baieli et al on lactoferrin purification from sweet whey. In reference to the time required for Lz adsorption, 4 h was chosen as the time for the adsorptive step, based on productivity terms (amount of purified protein per time and matrix amount) and as there was no significant difference ( P < 0.05), between 4 and 24 h using 100 mg matrix per 1 mL egg white (90.18% ± 1.54 and 96.36% ± 0.62, respectively).…”

“…In this sense, chitosan mini‐spheres were subjected to a first reaction with a bifunctional agent (epichlorohydrin) to crosslink the formed mini‐spheres, followed by a second reaction using an excess of epichlorohydrin to functionalize the chitosan hydroxyl and amino groups for immobilization of sulfanilic acid through its amine group (Figure ). Sulfanilic acid was chosen as ligand since this molecule is a precursor for the triazine dyes synthesis and our group has previously reported interesting results using them as ligands for affinity purification processes . The sulfanilic acid presents the advantage, over the triazine dyes, of not having regulatory restrictions if the matrix or product obtained from the process is used in the food industry.…”

“…Briefly, a chitosan solution was prepared by dissolving 2% of medium molecular weight chitosan powder in a 2% acetic acid solution . This solution was dripped through a 15G needle on a 2 M NaOH solution with soft continuous stirring, resulting in 1.60–1.80 mm size chitosan mini‐spheres . Sixteen hours later, the matrix was rinsed with distilled water until a neutral pH value.…”

“…Whereas chitin is insoluble in common solvents at any pH value, chitosan, the total or partial N ‐deacetylated form of chitin, is soluble in aqueous acid solutions. Due to its free amino and hydroxyl groups in its structure, chitosan is susceptible to undergo chemical modifications . Both chitin and chitosan present the advantage of being natural, biodegradable and environmentally safe biopolymers .…”

Molecular cloning, expression and purification of lactoferrin from Tibetan sheep mammary gland using a yeast expression system

“…Use of the yeast Pichi pastoris as an 69 expression system has several advantages including rapid growth 70 rate, ease of manipulation and high expression levels [11]. The 71 many purification procedures that have been used to isolate LF 72 include affinity chromatography [12], ion-exchange chromatogra-73 phy [13,14], batch extraction [15], and ultrafiltration on mem-74 branes [16]. However, a method with potential for use at an 75 industrial level of production is not yet available.…”