Process evaluations and economic analyses of recombinant human lysozyme and hen egg‐white lysozyme purifications

Abstract

Human lysozyme and hen egg-white lysozyme have antibacterial, antiviral, and antifungal properties with numerous potential commercial applications. Currently, hen egg-white lysozyme dominates low cost applications but the recent high-level expression of human lysozyme in rice could provide an economical source of lysozyme. This work compares human lysozyme and hen egg-white lysozyme adsorption to the cation exchange resin, SP-Sepharose FF, and the effect of rice extract components on lysozyme purification. With one exception, the dynamic binding capacities of human lysozyme were lower than those of hen egg-white at pH 4.5, 6, and 7.5 with ionic strengths ranging from 0 to 100 mM (5-20 mS). Ionic strength and pH had a similar effect on the adsorption capacities, but human lysozyme was more sensitive to these two factors than hen egg-white lysozyme. In the presence of rice extract, the dynamic binding capacities of human and hen egg-white lysozymes were reduced by 20-30% and by 32-39% at pH 6. Hen egg-white lysozyme was used as a benchmark to compare the effectiveness of human lysozyme purification from transgenic rice extract. Process simulation and cost analyses for human lysozyme purification from rice and hen egg-white lysozyme purification from egg-white resulted in similar unit production costs at 1 ton per year scale.

Cloning, Characterization, and Production of a Novel Lysozyme by Different Expression Hosts

“…Industrial production of hen egg white lysozyme benefits large-scale purification of thousands of kilogram of chicken eggs [31]. However, the production of natural lysozymes in bacteria is limited by the inefficient protein expression system.…”

“…Bacteriophage T4 lysozyme has been purified using the Ni-chelate affinity chromatography technique from overexpressing Escherichia coli cells [6]. Hen egg white lysozyme is produced at the metric ton scale by purification from eggs, and recent advances in transgenic plant technologies have led to similarly low production costs for recombinant human lysozyme [31]. Human lysozyme has also been expressed in Escherichia coli [16], Saccharomyces cerevisiae [12], and Pichia pastoris [28].…”

Engineering Escherichia coli for soluble expression and single step purification of active human lysozyme

“…Applications of wild type HEWL and hLYZ benefit from economies of scale, as both enzymes can be produced by the hundreds or thousands of kilograms via purification from chicken eggs or from recombinant rice, respectively (Wilken and Nikolov, 2011). In contrast, the study of other natural lysozymes or engineered variants can be stalled at preliminary stages due to limited material accessibility.…”

“…HEWL is produced at the metric tonne scale via purification from eggs, and recent advances in transgenic plant technologies have resulted in similarly low production costs for recombinant human lysozyme (hLYZ) (Wilken and Nikolov, 2011). These wild type enzymes thus benefit from economies of scale, but the native proteins suffer from inherent limitations with respect to some emerging applications, e.g.…”

Hyphenated Analytical Methods in Determination of Biologically Active Compounds in Hen's Eggs

“…Fat Folch, et al, 1957;Lima, 2002;Lisa, et al, 2011;Mazzella, et al, 2004;Palacios, et al, 2005;Romans, et al, 1972 Fatty Lysozyme Lyzes some bacteria antibacterial, antiviral, and antifungal agent Wang, et al, 2005;CegielskaRadziejewska, et al, 2008;Cunningham, et al, 1991;Wilken, et al, 2011 Ovoinhibitor Inhibits serine proteses antimicrobial potential Bourin, et al, 2011 Ovostatin Strongly antigenetic serine, cysteine, thiol, and metallo protease inhibitor Nain, et al, 2012 Cystain…”