Recombinant Rhodopirellula baltica UPF0314 protein RB9128 (RB9128)

Overview of Recombinant Rhodopirellula baltica UPF0314 Protein RB9128 (RB9128)

Rhodopirellula baltica UPF0314 protein RB9128 (RB9128) is a protein derived from the marine bacterium Rhodopirellula baltica . R. baltica is known for its distinct morphological properties and its role in the remineralization of biomass in marine environments . RB9128 is annotated as a protein of unknown function (UPF0314), which means that while its gene sequence is known, its specific biological role has not yet been fully characterized through experimentation .

Recombinant RB9128 is produced using genetic engineering techniques where the gene encoding RB9128 is inserted into a host organism (e.g., E. coli) to produce the protein in large quantities . The recombinant protein often includes an N-terminal His tag to facilitate purification .

Key Characteristics:

• Source: Rhodopirellula baltica

• Tag: His-tagged

• Molecular Weight: approximately 22.7 kDa [Calculated from AA sequence]

• Amino Acid Sequence: MNASSSETLEVVDDRFQKDRSWTVAWIAGLIVVGMVLVLAGMGRRFWCECGSWVPWSWDIWTAHNSQHLIDPYFFSHVLHGVLFYWALRWVPRLNRTQCLLIALGLEASWEILENSPLIIERYREATMAVGYTGDSIANSVTDVAACMLGYWFSSRFPWRWSVALFVVSEILMLIMIRDNLLLNVLMLVSPIPAIQEWQSG

• Purity: Greater than 90% as determined by SDS-PAGE

• Storage: Store at -20°C/-80°C upon receipt, avoid repeated freeze-thaw cycles .

• Reconstitution: Reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL .

Gene Information

The gene encoding RB9128 is designated as RB9128 . Synonyms for this gene include RB9128 and UPF0314 protein RB9128 . The UniProt ID for RB9128 is Q7UM18 .

Expression and Purification

Recombinant RB9128 is typically expressed in E. coli and includes a His-tag for purification using affinity chromatography .

Purification Steps:

1. Cell Lysis: E. coli cells expressing the recombinant protein are lysed to release the protein .

2. Affinity Chromatography: The lysate is passed through a chromatography column containing a resin that binds to the His-tag .

3. Washing: The column is washed to remove unbound proteins .

4. Elution: The bound protein is eluted from the column using a high concentration of imidazole, which competes with the His-tag for binding to the resin .

5. Buffer Exchange and Concentration: The eluted protein is then subjected to buffer exchange and concentration steps to achieve the desired buffer conditions and protein concentration .

Function and Regulation

While the precise function of RB9128 is currently unknown, proteomic studies of R. baltica have provided some context . Proteomic analyses have identified changes in protein composition during different growth phases and under various growth conditions .

R. baltica regulates its protein composition in response to different growth phases, with the number of regulated proteins increasing from early to late stationary growth phase . In stationary phase, several proteins of unknown function were up- or downregulated, suggesting their involvement in adaptation to changing environmental conditions .

Role in Carbohydrate Metabolism

R. baltica is a model organism for aerobic carbohydrate degradation in marine systems . Proteomic analysis has identified almost all enzymes of glycolysis, the TCA cycle, and the oxidative branch of the pentose phosphate cycle in R. baltica . These enzymes play crucial roles in the catabolism of various carbohydrates .