Difference between revisions of "Part:BBa K4275012"

Revision as of 10:09, 10 October 2022

CipA1B2C

CipA1B2C, a cellulosome integrating protein A with one integral carbohydrate-binding module 3 (CBM3), two type I cohesins, and a type II dockerin, is the primary scaffoldin of a cellulosome complex. The CBM3 binds to a range of polysaccharides, including cellulose and PET, increasing the catalytic efficiency of the enzymes. Meanwhile, the type I cohesins interact with the type I dockerins of cellulosomal enzymes, enabling the enzymes to attach to CipA1B2C. CipA1B2C forms a complete cellulosome complex when it binds to outer layer protein B (OlpB), an anchoring protein attached to a cell surface. CipA1B2C improves the efficiency of cellulosomal enzymes by increasing binding affinity to cellulose and PET through CBM and facilitating the spatial proximity among various enzymes, which allows enzymes to work in synergy. The ability to bind to various cellulosomal enzymes makes CipA a versatile tool for enhancing the efficiency of enzymes and conducting different enzyme synergies, which means CipA can promote the degradation rate of diverse materials (i.e. PET and cellulose). 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

CipA1B2C is a scaffold protein with one carbohydrate-binding module 3 (CBM3) and two type I cohesins. Natural CipA is found in Clostridium thermocellum, a gram-positive thermophilic and anaerobic bacterium. Cellulosomal enzymes bind tightly to CipA1B2C via the high-affinity dockerin I: cohesin I noncovalent interaction. CipA1B2C is then attached to OlpB, the secondary scaffoldin of the cellulosome complex, through the interaction between the type II dockerin and type II cohesin. OlpB is anchored to the cell surface. The CipA1B2C, OlpB, and the cellulosomal enzymes together 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 9 to 2, and the number of CBM is cut from 2 to 1 to reduce cellular pressure and ensure expression.

Sequence and Features