Difference between revisions of "Part:BBa K2380044"

 
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===Usage and Biology===
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<h2>Usage and Biology</h2>
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<p>
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In order to bring more variability in produced chitosans, we decided to implement a second chitin deacetylase. Because of its bacterial origin, we picked COD isolated from the gram-negative organism <i>Vibrio cholerae</i>. The <i>cod</i> gene is 1296 base pairs long and translates into a hydrolase with a molecular weight of approximately 45,5 kDa [2].
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Deacetylases target different units in a chitin molecule. Which unit is deacetylated depends on the chosen enzyme. In the case of COD, the second position from the non-reducing end is deacetylated [2][3]. The enzyme works optimally in surroundings with a pH of 8 and temperatures reaching 45 degrees Celsius [2][3].
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It is already proven that more than one CDA can be expressed in the same organism [2].
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In contrast to NodB CDA, COD does not deacetylate chitosan twice, after long incubation periods [2].
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CODs catalytic part is its N-terminal domain, while the other two domains make up carbohydrate-binding molecules. The catalytic domain correlates to a carbohydrate esterase domain (CDA) [3].
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</p>
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<h2>References</h2>
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<p>
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[1] National Center for Biotechnology Information, U.S. National Library of Medicine (NCBI); Vibrio cholerae O1 biovar eltor str. N16961 chromosome I, complete sequence, GenBank: AE003852.1; https://www.ncbi.nlm.nih.gov/nuccore/AE003852.1?from=1355388&to=1356683&sat=4&sat_key=105780702; last visited: 09/01/2017<br>
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[2] Hamer, S.N. et.al. Enzymatic production of defined chitosan oligomers with a specific pattern of acetylation using a combination of chitin oligosaccharide deacetylases (2015); Sci. Rep. 5, 8716; DOI:10.1038/srep08716<br>
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[3] Andrés, E. et.al., Albesa-Jové, D., Biarnés, X., Moerschbacher, B.M., Guerin, M., Planas, A. (2014) Structural Basis of Chitin Oligosaccharide Deacetylation; Angewandte Chemie International Edition, 53, 6882-6887; DOI: 10.1002/anie.201400220<br>
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[4] Li, X., Wang, L., Wang, X., Roseman, S. (2007) The Chitin Catabolic Cascade in the Marine Bacterium Vibrio Cholerae: Characterization of a Unique Chitin Oligosaccharide Deacetylase, Glycobiology, vol. 17, Issue 12, 1377–1387; DOI: 10.1093/glycob/cwm096
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</p>
  
 
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Revision as of 13:29, 24 October 2017


Chitin deacetylase COD

Chitin deacetylase (CDA) from Vibrio cholerae. The enzyme is a hydrolase and deacetylates chitin to chitosan. It solely deacetylates the second position of the non-reducing end in a chitin oligomer. We used it for the production of chitosan with a defined deacetylation pattern. It is not part of the pathogenic activity of Vibrio cholerea.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]