Part:BBa_K4349001
CfcI exochitinase from Aspergillus niger
CfcI is an exochitinase derived from the fungus Aspergillus niger.
Chitinases are enzymes that cleave chitin, a linear polysaccharide of N-acetyl-D-glucosamine units present in insect exoskeletons, fungal cell walls, cuticles and eggshells of nematodes. Chitinases are found in many organisms, including fungi, viruses, bacteria, insects, plants, and animals (Wang et al., 2007). These enzymes are classified as endochitinases, which cleave the polysaccharide randomly, and exochitinases, which cleave the ends (Seidl, 2008).
This part does not have a stop codon because we used it to assemble Composite Parts.
Usage and Biology
In the fungus, its expression was detected after the exponential growth phase, suggesting that its physiological function occurs in the late stages of the Aspergillus life cycle, such as autolysis or sporulation. A biochemical characterization study showed that CfcI exhibits optimal activity at pH 4 and in the temperature interval between 55–65 °C. It degrades chitin by cleaving and releasing N-acetylglucosamine from the reducing ends. This chitinase exclusively releases monomers (Munster et al., 2012).
The catalytic domain of CfcI consists of a triosephosphate isomerase barrel where a small additional (α+β) domain is inserted. In the (α+β) domain, next to the substrate binding cleft, there is a carbohydrate-binding module (CBM18) (Munster et al., 2012).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 518
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 105
Characterization
Construction:
To express the exochitinase PC, UFMG_UFV_Brazil Team synthesized the DNA sequence using the IDT grant. We received our parts, added an adenine overhang to the 3' extremities and successfully inserted our construction in TOPO vector via TA cloning.
References
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