C.Albicans PLB1
Escherichia Coli.
Protein is >90% pure as determined by 10% PAGE (coomassie staining).
Description
Recombinant Candida Albicans Phospholipase B1 (24-526 a.a) produced in E. coli having a Mw of 52kDa. C.Albicans PLB1 is fused to a 6xHis tag at its C terminal is and purified by proprietary chromatographic technique.
Product Specs
Escherichia Coli.
Product Science Overview
Phospholipase B1 (PLB1) is an enzyme that has been identified as a key factor in the pathogenicity of Candida albicans. It is believed to play a crucial role in the early stages of host invasion by the fungus. Due to its significant role in the infection process, PLB1 is considered a potential marker for diagnosing Candida albicans infections .
To study PLB1 in detail, researchers have expressed the enzyme recombinantly using an Escherichia coli expression system. This approach allows for the production of large quantities of the enzyme for further analysis. The recombinant PLB1 is initially found in inclusion bodies, which are aggregates of insoluble protein. These inclusion bodies can constitute up to 38.4% of the total insoluble protein produced in the bacterial cells .
The purification process of recombinant PLB1 involves several steps. Initially, the inclusion bodies are solubilized and the protein is refolded in a redox system containing reduced and oxidized glutathione (GSH/GSSG). The refolded protein is then purified using GST-sepharose 4B affinity chromatography, which exploits the GST-tag attached to the protein. After purification, the GST-tag is cleaved off using thrombin, and the protein is further purified by anion-exchange chromatography and reverse-phase high-performance liquid chromatography (HPLC) .
The final yield of purified PLB1 is approximately 15.6 milligrams from 100 milliliters of bacterial cell culture, with a concentration of 784 micrograms per microliter. The purified recombinant PLB1 exhibits phospholipase activity, as evidenced by the formation of a white precipitation zone on an egg yolk agar plate. The enzyme’s maximum activity is observed at a pH of 6.0 and a temperature of 37°C, with an activity level of 68 international units per milligram .
Recombinant PLB1 has several potential applications. It can be used in structural analytical studies to better understand the enzyme’s function and mechanism of action. Additionally, due to its role in the pathogenicity of Candida albicans, recombinant PLB1 can be utilized in diagnostic assays to detect infections caused by this fungus .
