Part:BBa_K3739069
J23100-B0030-his-CBM-hutH
His-tag enable us to purify the linking protein, CBM can bind to cellulose and hutH is a histidine transaminase which converts histidine into urocanic acid.
Biology
CBM
Cellulose enzymes have two domains, and the one that helps bind to cellulose is called cellulose binding module (CBM), and therefore it helps our fusion protein bind to cellulose-rich cell wall. Here we choose the CBM of CenA from Cellulomonas fimi, which has been successfully expressed in Escherichia coli.
hutH
The HutH comes from Pseudomonas putida. Under natural conditions, many microorganisms can use the histidine ammonia-lyase (HutH) to change L-histidine into urocanic acid. HutH catalyzes the first step in the degradation of histidine, and the product urocanic acid is further metabolized to glutamate. This enzyme could be found in the liver of vertebrates and in bacteria such as Escherichia coli, Salmonella and Pseudomonas. It is specific for L-histidine and can be inhibited by D-histidine or imidazole. The active center of the enzyme is thought to be dehydroalanine.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 607
Illegal NgoMIV site found at 1043
Illegal NgoMIV site found at 1778
Illegal NgoMIV site found at 2014
Illegal AgeI site found at 353
Illegal AgeI site found at 443
Illegal AgeI site found at 680
Illegal AgeI site found at 1507
Illegal AgeI site found at 2003 - 1000COMPATIBLE WITH RFC[1000]
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