Difference between revisions of "Part:BBa K3739011"

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===Biology===
 
===Biology===
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LMT
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=====LMT=====
 
Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in ''E.coli'': the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of ''Vibrio natriegens''. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of ''Vibrio natriegens''.
 
Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in ''E.coli'': the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of ''Vibrio natriegens''. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of ''Vibrio natriegens''.
  
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CBM
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=====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''.
 
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''.
  
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hutH
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=====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.
 
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.

Revision as of 23:43, 21 October 2021


LMT-his-CBM-hutH

LMT here represents a signal peptide used to secrect the fusion protein outside the cell. The fusion protein CBM-hutH works on the surface of single cell Phaeocystis. globosa and the enzyme catalyzes the reaction of converting histidine to form toxic urocanic acid. His-tag is added to purify the protein.


Biology

LMT

Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in E.coli: the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of Vibrio natriegens. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of Vibrio natriegens.

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.

Usage

Used to construct the composite part BBa_K3739039 and BBa_K3739109.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 597
    Illegal NgoMIV site found at 1033
    Illegal NgoMIV site found at 1768
    Illegal NgoMIV site found at 2004
    Illegal AgeI site found at 343
    Illegal AgeI site found at 433
    Illegal AgeI site found at 670
    Illegal AgeI site found at 1497
    Illegal AgeI site found at 1993
  • 1000
    COMPATIBLE WITH RFC[1000]