Difference between revisions of "Part:BBa K2380005"
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===Usage and Biology===
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<h2>Usage and Biology</h2>
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<p>Chitin is a molecule found in fungal cell walls [1]. Many plants possess enzymes, so-called chitinases, which are able to break down chitin and thus help along with its digestion. These enzymes play a role in defense mechanisms of plants, in case of fungal infections [2]. Even in human tissues chitinases appear, where they defend us against parasites [3]. Chitinases break down the glycosidic bonds between chitin monomer units, and are classified as hydrolases. <br>
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Its enzymatic activity has also previously been tested on chitin pentamers. ChiA1 breaks down these chitin pentamers in two dimers and one N-acetylglucosamine (GlcNAc) unit [4].<br>
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The N-terminal domain in the ChiA1 is responsible for its catalytic activity. The C-terminal domain plays an important role in the hydrolysis of chitin, and is the reason ChiA1 has such a high affinity to the substrate[4].
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</p>
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<h2>Results</h2>
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https://static.igem.org/mediawiki/2017/a/af/T--TU_Darmstadt--Chitinase_Page.png
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<p><b> Fig. 5: SDS-Page </b>
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<br>Lane 1: 24 hours after induction 
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<br>Lane 2: 3 hours after induction
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<br>Lane 3: Before induction
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<br>Lane 4: Empty BL21 cells</p>
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<p>To verify if ChiA1 is produced in BL21 cells containing the pSB1C3-AraC-chiA, a SDS-Page was performed.
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As we have described before, the page shows the expected results and proves that ChiA1 was successfully produced in <i>E.coli</i> BL21 cells.</p>
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<h2>References</h2>
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<p>
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[1] Stefanie Nicole Hamer, Stefan Cord-Landwehr, Xevi Biarnés, Antoni Planas, Hendrik Waegeman, Bruno Maria Moerschbacher, and Staphan Kolkenbrock (2015) Enzymatic production of defined chitosan oligomers with a specific pattern of acetylation using a combination of chitin oligosaccharide deacetylases
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DOI: 10.1038/srep08716 <br>
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[2] John G. Verburg and Q. Khai Huynh (1990) Purification and Characterization of an Antifungal Chitinase from Arabidopsis thaliana. Plant Physiol. 95, 450-455
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DOI: https://doi.org/10.1104/pp.95.2.450 <br>
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[3] Paoletti MG, Norberto L., Damini R., and Musumeci S. (2007) Human gastric juice contains chitinase that can degrade chitin
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DOI: 10.1159/000104144 <br>
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[4] Sylvain Cottaz, Eric Samain (2005) Genetic engineering of Escherichia coli for the production of NI,NII-diacetylchitobiose (chitinbiose) and its utilization as a primer for the synthesis of complex carbohydrates. Metabolic Engineering 7, 311–317 <br>
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</p>
  
 
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Revision as of 13:46, 28 October 2017


Chitinase A1

Chitinase A1 (ChiA1) is a hydrolase isolated from the organism Bacillus circulans WL-12. The chiA gene is 1983 base pairs long and translates into an enzyme with a molecular weight of approximately 69 kDa. ChiA1's main function is to break down glycosidic bonds between N-Acetylglucosamine (GlcNAc) units in a chitin oligomer.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 150
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 256
    Illegal AgeI site found at 952
    Illegal AgeI site found at 1240
    Illegal AgeI site found at 1303
    Illegal AgeI site found at 1366
    Illegal AgeI site found at 1696
  • 1000
    COMPATIBLE WITH RFC[1000]