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Mempro™ Cell-Free Alpha/Beta-Hydrolase Production

Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure now provides advanced custom Mempro™ protein production services for alph/beta-hydrolases based on cell-free expression system. You can count on us all through your projects.

Mempro™ cell-free protein production system is the innovative system to produce integral membrane proteins, which bypasses the limitations that encountered in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification using detergents. The alpha/beta-hydrolase family is a large class of structurally related enzymes with diverse catalytic functions. Enzymes in this family contain a specific domain termed as alpha/beta-hydrolase fold, which is an alpha/beta-sheet containing 8 beta strands connected by 6 alpha helices. The alpha/beta-hydrolase fold mainly exsit in protein members such as esterases, lipases, proteases, peroxidases, epoxide hydrolases and dehalogenases.

Figure 1. Schematic diagram of alpha/beta-hydrolase fold.

Creative Biostructure has extensive expertise in high-yield alpha/beta-hydrolase production using cell-free protein expression system, we can provide various strategies for Mempro™ cell-free protein production, including:

Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is commonly used for alpha/beta-hydrolase production. Wheat germ extract can provide eukaryotic translation system for alpha/beta-hydrolase expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for alpha/beta-hydrolase expression. Notably, crude rabbit reticulocyte lysate is another powerful tool for eukaryotic cell-free membrane protein expression. To select an appropriate source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.

With the Mempro™ cell-free protein production platform, Creative Biostructure is able to express, purify and crystallize alpha/beta-hydrolases to facilitate the study of their biological functions. Mempro™ cell-free membrane protein production has a large of advantages, including:

  • Very high yield;
  • Low cost in time and money;
  • Lack of toxicity;
  • Incorporation of non-natural amino acids into protein structure;
  • Relatively tolerant of additives.

Most notably, One challenge associated with Mempro™ cell-free membrane protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize Mempro™ cell-free protein production for alpha/beta-hydrolases using plasmids due to they are circular and have no end for the exonucleases to attach.

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

References:
Alpha/beta hydrolase fold. (https://en.wikipedia.org/wiki/Alpha/beta_hydrolase_fold).
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
M. Holmquist (2000). Alpha/beta-hydrolase fold enzymes: structures, functions and mechanisms. Curr. Protein Pept. Sci., 1(2): 209-35.
N. Lenfant, et al. (2012). ESTHER, the database of the α/β-hydrolase fold superfamily of proteins: tools to explore diversity of functions. Nucl. Acids. Res., 41: D423-429.


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