Difference between revisions of "Part:BBa K5117005"

Revision as of 18:51, 29 September 2024

AtCelO

This part contains the celO gene of Acetivibrio thermocellus (synonym Clostridium thermocellum) including its native signal peptide for secretion, encoding an exoglucanase (EC 3.2.1.176).

Sequence and Features

Enzyme characterization according to literature

In the study of Zverlov et al. titled “A newly described cellulosomal cellobiohydrolase, CelO, from Clostridium thermocellum: investigation of the exo-mode of hydrolysis, and binding capacity to crystalline cellulose”, the structure of the celO gene from Clostridium thermocellum F7 is reported and the corresponding protein was shown to possess cellobiohydrolase activity (Zverlov et al. 2002).

Two truncated proteins were constructed and examined: rCelO, with the leader peptide and the dockerin module deleted (587 aa, 67.3 kDa), and rCelO-Cat, representing only the catalytic domain of CelO (415 aa, 47.9 kDa). The resulting enzymes were recombinantly produced in Escherichia coli and purified via 6x-His tag purification method. Using barley β-glucan as substrate, the optimal temperature and pH were determined to be 65 °C and 6.6, respectively (Zverlov et al. 2002).

More information related to this part can be found in the following publications and databases:

• Ahmad S., Sajjad M., Altayb H. N., Sarim Imam S., Alshehri S., Ghoneim M. M., Shahid S., Mirza M. U., Nadeem M. S., Kazmi I., Waheed Akhtar, M. (2023): Engineering processive cellulase of Clostridium thermocellum to divulge the role of the carbohydrate‐binding module. Biotechnology and Applied Biochemistry 70(1), 290-305. https://doi.org/10.1002/bab.2352
• Gene sequence: https://www.ncbi.nlm.nih.gov/nuccore/AJ275975
• Protein sequence: https://www.ncbi.nlm.nih.gov/protein/CAB76938
• UniProtKB: https://www.uniprot.org/uniprotkb/Q9L3J2/entry

References

Zverlov V. V., Velikodvorskaya G. A., Schwarz W. H. (2002): A newly described cellulosomal cellobiohydrolase, CelO, from Clostridium thermocellum: investigation of the exo-mode of hydrolysis, and binding capacity to crystalline cellulose. Microbiology 148(1), 247-255. https://doi.org/10.1099/00221287-148-1-247