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Mempro™ Cell-Based Lipoxygenase Production

Creative Biostructure has built an first-class membrane protein production platform. Our custom Mempro™ lipoxygenase production service is based on our advanced cell-based expression systems, which performs the desired membrane protein production within a living cell.

Lipoxygenases, LOX for short, are a family found in a various organisms from the planta, animalia, bacteria to fungi. Lipoxygenases are dioxygenases containing iron or manganese. There are two groups of lipoxygenases, the plant lipoxygenases and the other lipoxygenases. Unsaturated fatty acids (UFA) can be conversed into the corresponding oxylipins by lipoxygenases. The primary lipoxygenases reaction is the production of hydroperoxides, which can be utilized in multiple industrial applications from food processing to oleochemical production. One of the potential renewable raw materials for the chemical industry is biomass derived lipids. Lipid derived hydroperoxides produced by lipoxygenases can be transformed into varouis functional oleochemicals precursors. In furture, lipoxygenases may play a crucial role in novel environmentally friendly processes design, especially in oleochemistry. So far, however, the major problem is insufficient lipoxygenases production for large-scale application.

Mempro™ Cell-Based Lipoxygenase Production
Figure 1. NMR structures of the Glycine max (A) and Pseudomonas aeruginosa (B) lipoxygenase. (Critical Reviews in Biotechnology, 2016)

Based on technology innovation, our cell-based expression platform is aimed to overcome the disadvantages in membrane protein production. Creative Biostructure could provide you the high-quality membrane protein production services, and we can perform varouis Mempro™ cell-based protein production systems including:

•      Mempro™ Lipoxygenase Production using Insect Cells System
The most applied system for eukaryotic membrane protein production is in cultured insect cells and larvae. Mempro™ protein production using insect cells system can increase the expression level and reduce the truncated proteins compared to E. coli system according to similar codon usage rules. Our innovational insect cells system based on baculovirus is advanced in several aspects as below:
Accuracy;
Easy scale-up;
High membrane protein yield;
Proper folding and post translation modifications;
Use of cell lines ideal for suspension culture.

•      Mempro™ Lipoxygenase Production using Bacterial Cells System
Escherichia coli (E. coli) is the most widely applied bacterial host for membrane protein production. Lemo21 (DE3) strain is optimized for our Mempro™ protein production in our bacterial cells system. The target membrane protein encoding gene is located on a pET vector governed by the T7lac promoter. Membrane proteins are expressed as C-terminal GFP fusion protein as a result.

•      Mempro™ Lipoxygenase Production using Mammalian Cells System
Creative Biostructure can provide Mempro™ protein production in mammalian cells system. This system faciliates membrane proteins to perform correctly folding and post-translational modification. Various applications for eukaryotic membrane protein production by Mempro™ mammalian cells system lead to the best structural and functional features.

      Mempro™ Lipoxygenase Production using Yeast Cells System
Single-celled yeast is an easy, quick and economic culture host and able to apply multiple post-translational modifications for eukaryotic membrane protein. The most widely used two species of yeast for membrane protein production are Pichia pastoris (P. pastoris) and Saccharomyces cerevisiae (S. cerevisiae). Creative Biostructure has developed several strategies to improve yields per cell through optimizing the expression plasmid, host cell and culture conditions.

Creative Biostructure conducts high-yield lipoxygenase production by Mempro™ cell-based protein production services including expression, isolation, purification and crystallization.

We provide an array of Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
R. Heshof, et al. (2016). Industrial potential of lipoxygenases. Mol. Pharmacol., 36(4): 665-674.
A. GarretaA, et. al. (2013).Structure and interaction with phospholipids of a prokaryotic lipoxygenase from Pseudomonas aeruginosa. FASEB. J, 27, 4811–4821.
J. J. Villaverde, et. al. (2012). Formation of oligomeric alkenylperoxides during the oxidation of unsaturated fatty acids: an electrospray ionization tandem mass spectrometry study. J. Mass Spectrom, 47, 163–172.


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