Difference between revisions of "Part:BBa K4275013"

Revision as of 14:26, 9 October 2022

OlpB-3C-Ag3

OlpB3CAg3, an outer layer protein B with 3 type II cohesins and an antigen 3 (Ag3), is the secondary scaffoldin of cellulosome complex. The main function of OlpB is securing cellulosome integrating protein A (CipA) to the cell surface. The type II cohesin interacts with the type II dockerin on CipA, binding CipA to OlpB. In turn, OlpB adheres to the cell surface through the interaction between Ag3 and nanobody 3 (Nb3) present on the cell surface. The binding to the cell surface effectively increases the efficiency of the cellulosome complex. This is a part in a part collection where we enable efficient degradation of cellulose and PET in textile waste.

The part collection includes:

Our part collection can help inspire future teams to design and perfect the degradation of cellulose, adding to the collection.

Usage and Biology

OlpB3CAg3 is the secondary scaffoldin of cellulosome complex that has 3 type II cohesins and an antigen 3 (Ag3). Natural OlpB is found in Clostridium thermocellum, a gram-positive thermophilic and anaerobic bacterium. OlpB is responsible for binding several primary scaffoldin of cellulosome complex, CipA, to the cell surface. OlpB binds to CipA via the interaction between the type II dockerin and type II cohesin. In nature, OlpB attaches to the cell surface of Clostridium thermocellum through the interaction between surface-like homologous (SLH) domains and the cell surface. Together, the CipA1B2C, OlpB, and the cellulosomal enzymes comprise a multiplex cellulosome. Utilizing the highly effective cellulosome complex, C. Thermocellum is the most efficient microorganism for lignocellulosic biomass degradation.

Design consideration

1. 6x histag (HHHHHH) added to C-terminal to allow for Ni-NTA purification.

2. The number of cohesins is cut from 7 to 3 to reduce cellular pressure, thus ensuring expression.

3. The original anchoring domain of OlpB that binds to yeast surface, glycosylphosphatidylinositol (ScGPI), is replaced by Ag3 so that OlpB can attach to the cell surface of E.coli through the interaction between Ag3 and Nb3.

Sequence and Features