Difference between revisions of "Part:BBa K1602018"
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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. | 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). | 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]