Coding
HisTag-HIC

Part:BBa_K1602018

Designed by: Sebastian Barthel, Jonas Sindlinger, Bianca Reisinger, Sven Jager   Group: iGEM15_TU_Darmstadt   (2015-09-16)

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]


[edit]
Categories
Parameters
None