Coding
BC2

Part:BBa_K3979003:Design

Designed by: Akshay J, Abhishek Raghunathan   Group: iGEM21_IISER_TVM   (2021-10-18)

Bacterial Chitinase Combo 2


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 7
    Illegal XhoI site found at 1156
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 254
    Illegal SapI.rc site found at 15

Design Notes

This is a chitinase gene designed using the catalytic domain(CD) of chitinase from Amycolatopsis orientalis and the chitin-binding domain (CBD) of ChiB from Serratia marcescens. The domains mentioned above are linked without any extra linking residues considering the fact that CBD of Streptomyces chitinase is not yet annotated, and CD-CBD makes a basic chitinase gene framework. The protein sequences were linked initially and then reverse translated to nucleotide sequences(stop codon inclusive) along with codon optimization for E.coli expression using the BOOST Juggler tool.BamHI and HindIII sites were added at the 5’ and 3’ end of the gene, respectively.


Source

Chitinase B Serratia marcescens GenBank ID: P11797

Chitinase of Amycolatopsis orientalis GenBank ID: WP_044850297.1


References


  • Streptomyces orientalis: Yoshio Tominaga & Yoshio Tsujisaka (1976) Purifications and Some Properties of Two Chitinases from Streptomyces orientalis Which Lyse Rhizopus Cell Wall, Agricultural and Biological Chemistry, 40:12, 2325-2333, DOI: 10.1080/00021369.1976.10862407
  • Serratia marcescens: Li J, Zheng J, Liang Y, Yan R, Xu X, Lin J. Expression and characterization of a chitinase from Serratia marcescens. Protein Expr Purif. 2020 Jul;171:105613. doi: 10.1016/j.pep.2020.105613. Epub 2020 Feb 23. PMID: 32097727
  • Saima, & Kuddus, Mohammed & Roohi, & Ahmad, Iffat. (2013). Isolation of novel chitinolytic bacteria and production optimization of extracellular chitinase. Journal of Genetic Engineering and Biotechnology. 11. 39–46. 10.1016/j.jgeb.2013.03.001
  • "Circular Dichroism", Wikipedia, 2021. Accessed 18 Oct 2021
  • "Circular Dichroism Spectroscopy", Cryst.Bbk.Ac.Uk, 2021, School of Crystallography, Department of Biological Sciences, Birkbeck. Accessed 18 Oct 2021
  • O. Trott, A. J. Olson, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading, Journal of Computational Chemistry 31 (2010) 455-461 DOI 10.1002/jcc.21334