Part:BBa_K3493000:Design
G. algens dextranase CDS
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 385
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 385
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 385
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 385
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part is the CDS of a dextranase from a psychrophile organism (Gelidibacter algens) codon optimized for expression in Escherichia coli. Following reports that the Streptococcus mutans homologous dextranase has a higher activity when its N terminal (residues 1-99) and C-terminal (residues 733-850) are deleted (Kim et al., 2011), we removed them from the CDS submitted to the registry. Accordingly, our final CDS contains the region of the G. algens dextranase sequence that aligns to residues 104 - 734 of the PDB structure of the S. mutans dextranase (Suzuki et al., 2012). The dextranase of G. algens was successfully expressed in E. coli using the pET28a expression vector.
Source
The protein sequence is available at NCBI (https://www.ncbi.nlm.nih.gov/protein/WP_066432599.1?report=genbank&log$=prottop&blast_rank=700&RID=MZBBN4NZ014). The gene sequence was inferred from the protein sequence using codon optimization for E. coli.
References
Kim, Y.-M., Shimizu, R., Nakai, H., Mori, H., Okuyama, M., Kang, M.-S., Fujimoto, Z., Funane, K., Kim, D., & Kimura, A. (2011). Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity. Applied Microbiology and Biotechnology, 91(2), 329–339.
Suzuki, N., Kim, Y.-M., Fujimoto, Z., Momma, M., Okuyama, M., Mori, H., Funane, K., & Kimura, A. (2012). Structural elucidation of dextran degradation mechanism by streptococcus mutans dextranase belonging to glycoside hydrolase family 66. The Journal of Biological Chemistry, 287(24), 19916–19926.