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Magic™ Membrane Protein Expression in Photosynthetic Bacteria

Creative Biolabs constantly strives to provide unparalleled membrane protein expression service using a variety of systems, particularly photosynthetic bacteria cells. Our scientists have accumulated a wealth of experience after long-term exploration, and successfully established and commercialized a world-leading Magic™ Membrane Protein Production Platform. We are determined to assist our worldwide clients to facilitate their research progress.

Membrane proteins comprise over 30% of all encoding genes in the entire human genome. They participate in various cellular events and manifest as the major class of therapeutic target. Therefore, further knowledge of the structures and functions of membrane proteins would contribute greatly to our understanding of cell biology as well as novel drug development. Unfortunately, structural information of membrane proteins is relatively scarce since it is difficult to purify sufficient quantities of native proteins for downstream applications, e.g. crystallization. Heterologous expression ( e.g. E. coli cells, yeast cells, baculovirus-insect cells and mammalian cells) are often cumbersome and expensive because of relatively low yield. Other limitations of these systems include saturation of the secretory machinery for integration of the heterologous protein into the membrane, and inadequate membrane volume for the accommodation of de novo synthesized proteins. Thus, overexpression strategies often result in cell death, aggregates, or misfolded/denatured protein products.

Photosynthetic bacteria (e.g. Rhodobacter) provides a promising alternative to this question since they hold a featured inducible intracytoplasmic membrane (ICM). Under conditions of light and/or lowered oxygen, the membrane surface of the native organism increases significantly when the ICM is elaborated as invaginations of the cytoplasmic membrane. Moreover, such environmental stimulations induce synthesis of the photosynthetic apparatus. The new ICM in the native organism sequesters these complexes that are composed of transmembrane polypeptides and their associated hydrophobic redox and energy transfer cofactors. Upon cell splitting, the ICM break apart from the host’s cytoplasmic membrane becoming sealed “inside-out” particles that can be easily isolated by differential centrifugation through their size. With the development of biologics, large quantities of active membrane proteins can be obtained. As such, photosynthetic bacteria system exerts great potential for both biotechnology and pharmaceutical industries.

Representative process of membrane protein expression using 80-ml photosynthetic bacteria cell cultures grown semi-aerobically Figure 1.  Representative process of membrane protein expression using 80-ml photosynthetic bacteria cell cultures grown semi-aerobically
(Production of Membrane Proteins: Strategies for Expression and Isolation, 2011)

The advantages of Magic™ protein expression in photosynthetic bacteria cells are described as below:


Creative Biolabs has extensive expertise in membrane protein expression. Our one-stop service can offer customers the most flexible options and most reliable outcomes. Please feel free to inquiry us for more information.

References

  1. Bernaudat F, Frelet-Barrand A, Pochon N, et al. (2011). Heterologous expression of membrane proteins: choosing the appropriate host. PloS one, 6(12), e29191.
  2. Laible P D, Mielke D L, Hanson D K. (2009). Foreign gene expression in photosynthetic bacteria. In The Purple Phototrophic Bacteria (pp. 839-860). Springer Netherlands.
  3. Laible P D, Mielke D L, Hanson D K. (2011). Membrane Protein Production Using Photosynthetic Bacteria: A Practical Guide. Production of Membrane Proteins: Strategies for Expression and Isolation, 167-198.



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