Recombinant Saccharum officinarum Photosystem II CP47 chlorophyll apoprotein (psbB)
Description
Definition and Biological Role
Recombinant Saccharum officinarum Photosystem II CP47 chlorophyll apoprotein (psbB) is a genetically engineered form of the intrinsic chlorophyll-binding protein critical for Photosystem II (PSII) function. CP47, encoded by the psbB gene, serves as a core antenna complex in PSII, facilitating light energy transfer to the reaction center and stabilizing the oxygen-evolving complex (OEC) . In sugarcane (Saccharum officinarum), recombinant expression enables biochemical and structural studies of this protein under controlled conditions.
Recombinant Production Methodology
Recombinant CP47 is typically produced via the following workflow:
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Gene Cloning: The psbB coding sequence (1–508 aa) is codon-optimized for E. coli and inserted into a plasmid .
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Transformation: Plasmid is introduced into E. coli strains (e.g., BL21) for heterologous expression.
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Induction and Purification: Protein expression is induced with IPTG, followed by affinity chromatography (Ni-NTA for His-tagged proteins) .
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Validation: SDS-PAGE confirms >90% purity, and mass spectrometry identifies co-purified subunits (e.g., PsbH, PsbL) .
Key Challenges:
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Solubility: Requires fusion tags (e.g., His-SUMO) and detergents for membrane protein stabilization .
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Pigment Retention: Chlorophyll incorporation in vitro often yields lower efficiency compared to in vivo assembly .
Functional Insights from Mutagenesis Studies
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Chloride Binding: The R448 residue in loop E is critical for chloride coordination. Mutants like R448S exhibit 50% reduced photoautotrophic growth and impaired OEC reactivation under chloride limitation .
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Photoinactivation: CP47 mutants show increased susceptibility to photodamage, highlighting its role in PSII repair cycles .
Applications in Photosynthesis Research
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PSII Assembly Studies: Recombinant CP47 aids in probing early PSII assembly intermediates, such as the "RC47" subcomplex with D1/D2 reaction centers .
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Antibody Development: Anti-CP47 antibodies (e.g., Agrisera AS05 084) enable quantitative PSII analysis in plants and algae .
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Biotechnological Engineering: Insights from CP47 structure inform the design of synthetic light-harvesting systems .
Limitations and Future Directions
While recombinant CP47 from Spinacia oleracea (spinach) and Draba nemorosa is well-characterized , Saccharum officinarum-specific studies remain scarce. Future work should prioritize:
Product Specs
Note: We will prioritize shipping the format currently in stock. However, if you have specific format requirements, please indicate them in your order notes, and we will fulfill your request as best as possible.
Note: All protein shipments are standardly packaged with blue ice packs. If you require dry ice shipping, please inform us in advance as additional fees will apply.
Generally, liquid forms have a shelf life of 6 months at -20°C/-80°C, while lyophilized forms have a shelf life of 12 months at -20°C/-80°C.
