Difference between revisions of "Part:BBa K1602018"
 
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<h1>Humicola insolens cutinase with N-terminal His-Tag</h1>
 
<h1>Humicola insolens cutinase with N-terminal His-Tag</h1>
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<b>cadA</b> is a gene, coding for a cis-aconitate decarboxylase in Aspergillus terreus. The decarboxylase catalyzes the conversion from cis-aconitate to itaconic acid. The reaction takes place in the cytosol of the host and frees one molecule of carbon dioxide per conversion.
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The Humicola insolens cutinase (HIC) (EC 3.1.1.74, cutinase) belongs to the serine-α -β-hydrolase family (Nielsen et al., 2005) and can be used to degrade polymers like polyethylene terephthalate (Espino-Rammer et al., 2013). The catalytic pocket consists of the triad of a serine, histidine and aspartic acid (Kold et al., 2014). In addition it was reported that HIC catalyze condensation and lactone ring-opening polymerizations in an organic phase (Hunsen et al., 2008) at an optimal activity of 70°C. Assays showed no ester cleavage activity after incubating at 70°C for 15 min.
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An N-terminal attached Polyhistag enables purification via immobilized metal affinity chromatography (IMAC).
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Experimental results are available in the [https://parts.igem.org/Part:BBa_K1602020 inducing operon of HIC].
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Latest revision as of 03:47, 19 September 2015

Humicola insolens cutinase with N-terminal His-Tag

The Humicola insolens cutinase (HIC) (EC 3.1.1.74, cutinase) belongs to the serine-α -β-hydrolase family (Nielsen et al., 2005) and can be used to degrade polymers like polyethylene terephthalate (Espino-Rammer et al., 2013). The catalytic pocket consists of the triad of a serine, histidine and aspartic acid (Kold et al., 2014). In addition it was reported that HIC catalyze condensation and lactone ring-opening polymerizations in an organic phase (Hunsen et al., 2008) at an optimal activity of 70°C. Assays showed no ester cleavage activity after incubating at 70°C for 15 min. An N-terminal attached Polyhistag enables purification via immobilized metal affinity chromatography (IMAC). Experimental results are available in the inducing operon of HIC.

Characteristics

Molecular Weight 21068.85
Residues 200
Charge 5.5
Isoelectric Point 8.1624
A280 Molar Extinction Coefficients 12950 (reduced) 13200 (cystine bridges)
Improbability of expression in inclusion bodies 0.891
[Data taken from PEPSTATS]

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]