Difference between revisions of "Part:BBa K3394000"

Revision as of 07:41, 15 October 2020

Coding Sequence of Endo5a cellulase (for E. coli)

This part consists the coding region of Endo5A, an endo-1,4-beta-glucanase. Endo5a breaks down cellulose by cutting internal 1-4 beta glucose linkages in the cellulose chain.

Figure 1: Cellulose chain. 1-4 beta glucose linkage is highlighted

The complete cellulose hydrolysis reaction involves this enzyme along with several other enzymes: endo-1,4-beta-glucanases, exo-1,4-beta-glucanases (or cellobiohydrolases) and beta-glucosidases.

Endo5a is a relatively small protein with an optimal temperature of 50 Celsius degrees and an optimal pH range of 6-7. The NCBI Genbank number for Endo5a is HQ657203.1 (DNA), AEB00655.1 (amino acid).

We verified that the sequence is optimized for use in Escherichia coli, and it was synthesized by IDT for us. Furthermore, Endo5a was expressed with an N-terminal histidine tag (6-histidine tag) instead of the 17 first amino acids that served as a signal peptide (51 nucleotides).

Endo5A was isolated from the bacterial flora found in the gut of a cotton bollworm (Helicoverpa armigera) which secretes a variety of plant-hydrolyzing enzymes.

Sequence and Features