Difference between revisions of "Part:BBa K1498000"
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<partinfo>BBa_K1498000 short</partinfo> | <partinfo>BBa_K1498000 short</partinfo> | ||
| − | + | BBa_K1498000 comprises of the (codon optimised for ''E. coli'') coding region of kerUS keratinase from ''Brevibacillus brevis''. kerUS harbours a signal peptide for extracellular secretion, which is cleaved following secretion -to yield an active keratinase (Jaouadi et al., 2013). | |
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| + | Keratins are a class of fibrous, insoluble and abundant structural proteins which are found in structures such as hair, feathers, nails, hooves and wool. these proteins are highly stable, showing resistance to many proteolytic enzymes. | ||
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| + | Keratinases are a group of proteases that can degrade keratin substrates. | ||
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| + | BBa_K1498000 is a 383aa, 39.4 kDa protein. There is a predicted a signal peptide of 29 aa, which directs the protein for secretion. Secretion was shown to be effective in ''E. coli'' in addition to the endogenous organism. This along with other residues from a pro-sequence consisting of 79 aa which are cleaved by autoproteolytic processing in the periplasm. The active mature keratinase consists of 275 aa, with a molecular weight of 29 kDa. | ||
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| + | '''References.''' | ||
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| + | Jaouadi, N. Z., Rekik, H., Badis, A., Trabelsi, S., Belhoul, M., Yahiaoui, A. B., … Jaouadi, B. (2013). Biochemical and molecular characterization of a serine keratinase from Brevibacillus brevis US575 with promising keratin-biodegradation and hide-dehairing activities. PloS one, 8(10), e76722. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | While it was reported that the processed protein had successfully been isolated following extracellular expression in ''E. coli'' BL21 this was not achieved by our team. Following expression and purification of the supernatant a protein of 39.4 kDa was observed on a coomassie stained SDS-PAGE gel. Furthermore following assays to measure keratinase activity, none was observed -implying that the pre-processed protein is inactive | ||
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Latest revision as of 18:33, 17 October 2014
kerUS keratinase from Brevibacillus brevis
BBa_K1498000 comprises of the (codon optimised for E. coli) coding region of kerUS keratinase from Brevibacillus brevis. kerUS harbours a signal peptide for extracellular secretion, which is cleaved following secretion -to yield an active keratinase (Jaouadi et al., 2013).
Keratins are a class of fibrous, insoluble and abundant structural proteins which are found in structures such as hair, feathers, nails, hooves and wool. these proteins are highly stable, showing resistance to many proteolytic enzymes.
Keratinases are a group of proteases that can degrade keratin substrates.
BBa_K1498000 is a 383aa, 39.4 kDa protein. There is a predicted a signal peptide of 29 aa, which directs the protein for secretion. Secretion was shown to be effective in E. coli in addition to the endogenous organism. This along with other residues from a pro-sequence consisting of 79 aa which are cleaved by autoproteolytic processing in the periplasm. The active mature keratinase consists of 275 aa, with a molecular weight of 29 kDa.
References.
Jaouadi, N. Z., Rekik, H., Badis, A., Trabelsi, S., Belhoul, M., Yahiaoui, A. B., … Jaouadi, B. (2013). Biochemical and molecular characterization of a serine keratinase from Brevibacillus brevis US575 with promising keratin-biodegradation and hide-dehairing activities. PloS one, 8(10), e76722.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 866
- 1000COMPATIBLE WITH RFC[1000]