Mempro™ Cell-Free Lipoxygenase Production

Creative Biostructure offers advanced custom Mempro™ protein production services for lipoxygenase (LOX) using cell-free expression system.

Mempro™ cell-free protein production system is a cutting-edge system developed by Creative Biostructure for efficient production of membrane proteins. It has unique advantages over the traditional cell-based protein production system, such as less concern of toxicity, high yield, post-translational modifications included etc.

Figure 1. Structure of rabit 15-lipoxygenase

Lipoxygenase consists of a structural related family of iron-containing enzymes that catalyze the addition of oxygen to esstial polyunsaturated fatty acids containing a cis,cis-1,4- pentadiene structure in human. LOX could be found in plants, fungi, mammals, invertebrates and bacteria. Their functions include initiating the signaling molecule synthesis and inducing the structural or metabolic changes in the cell. Members of LOX proteins contain highly conserved domains and sequence motifs that are responsible for their distinct structure and the interaction with the catalytic iron, for example the “LOX motif”: His-X4-His-X4-His-X17-His-X8-His. In mammalian cells and plant cells, LOX consists of two domains: a N-terminal β-barrel domain named PLAT (Polycystin-1, lipoxygenase, Alpha-Toxin) and a C-terminal catalytic domain LH2 (Lipoxygenase homology) containing a single atom of non-heme iron. This iron atom is bound to conserved histidines and to the carboxyl group of a conserved C-terminal isoleucine.

Creative Biostructure is very experienced in high-yield lipoxygenase production using cell-free protein expression systems, various strategies are available for Mempro™ cell-free protein production, including:

• Mempro™ Cell-Free Protein Production in E. coli;
• Mempro™ Cell-Free Protein Production in Wheat Germ;
• Mempro™ Cell-Free Protein Production in Rabbit Reticulocyte;
• Mempro™ Cell-Free Protein Production in Baculovirus.

Among Mempro™ Cell-Free Protein Production systems, E. coli S30 extract system is widely used for its economical and rapid functional protein production. Wheat germ extract system is a highly stable and robust system which contains the essential eukaryotic translation system for lipoxygenase production. Another major system for lipoxygnase production is Baculovirus expression system that derived from Autographa californica nucleopolyhedrovirus (AcNPV). It’s particularly suitable for large scale application. In addition, the Crude rabbit reticulocyte lysate is a widely used RNA-dependent system, since it has low background and requires minimal amount of RNA.

With the Mempro™ cell-free protein production platform, Creative Biostructure could express, purify and crystallize lipoxygenase in an efficient and effective manner to facilitate the study of their biological functions.

We provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
Gillmor, Sarah A., et al. "The structure of mammalian 15-lipoxygenase reveals similarity to the lipases and the determinants of substrate specificity."Nature Structural & Molecular Biology 4.12 (1997): 1003-1009.
Brash A R. Lipoxygenases: occurrence, functions, catalysis, and acquisition of substrate[J]. Journal of Biological Chemistry, 1999, 274(34): 23679-23682.