Part:BBa_K200007
Cellulase
This is the coding sequence for [http://en.wikipedia.org/wiki/Cellulase cellulase] production in E.coli. This biobrick codes for endo-b-1,4-glucanase E (CelE) cellulase, which is one of the three major proteins of the cellulosome of Clostridium cellulolyticum.1
Cellulase mainly catalyses the reactions that changes crystalline cellulose to cellobiose2 and then finally to glucose. It also catalyses, to a small extent, the break down of carboxymethyl cellulose.
This cellulase is protease resistant.
As part of the Imperial 2009 iGEM E.ncapsulator project, E.coli was engineered to synthesise cellulase to a tunable threshold. Following that, E.coli is to be self-encapsulated so as to protect cellulase through the stomach, until its release in the small intestine. Cellulase will then be able to digest the cellulose found in our food.
Usage and Biology
It can be used in gram positive bacteria.
The enzyme has a C-end cellulosome-binding domain (CBD), which is used to deliver its resident catalytic domain to the cellulosome. The cellulosome is multi-subunit complex that is involved in the hydrolysis of crystalline cellulose. [http://www.springerlink.com/content/lp8g657n13g76187/fulltext.pdf [3]]
In Clostridum, CelE activity activates other cellulosomal enzymes synergistically.2
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 199
Illegal PstI site found at 700 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 700
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 287
Illegal BamHI site found at 814
Illegal XhoI site found at 524 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 199
Illegal PstI site found at 700 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 199
Illegal PstI site found at 700 - 1000COMPATIBLE WITH RFC[1000]
References
[http://www.ncbi.nlm.nih.gov/nuccore/125972525?report=genbank&log$=seqview&from=961597&to=964041 Sequence from NCBI] <biblio>
- 1 pmid=9055408
- 2 pmid=10714996
</biblio>
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