Difference between revisions of "Part:BBa K4719011"
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==Usage and biology==
 
==Usage and biology==
 
AnCDA is chitin deacetylase from ''Aspergilus nidulans'' FGSCA4. This deacetylase is a member of carbohydrate esterase family 4 (CE4) of the CAZy database ([http://www.cazy.org/, www.cazy.org]). It has broad substrate specifity – AnCDA hydrolyzes N-acetamido groups in chitin oligomers, crystalline chitin and chitosan, however, it does not deacetylase peptidoglycan. AnCDA reaches its highest level of activity at approximately 50°C and pH 8.0 when Co2+ is used as a cofactor.
 
AnCDA is chitin deacetylase from ''Aspergilus nidulans'' FGSCA4. This deacetylase is a member of carbohydrate esterase family 4 (CE4) of the CAZy database ([http://www.cazy.org/, www.cazy.org]). It has broad substrate specifity – AnCDA hydrolyzes N-acetamido groups in chitin oligomers, crystalline chitin and chitosan, however, it does not deacetylase peptidoglycan. AnCDA reaches its highest level of activity at approximately 50°C and pH 8.0 when Co2+ is used as a cofactor.
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AnCDA gene has three exons and encodes the primary product of 237 amino acids, including the N-terminal extracellular signal sequence. Coding regions except leader sequence were cloned into pBAD/HisB vector for recombinant protein expression in E. coli and protein purification using Ni-NTA chromatography with His-tag fused to AnCDA in N-terminal.
 
AnCDA gene has three exons and encodes the primary product of 237 amino acids, including the N-terminal extracellular signal sequence. Coding regions except leader sequence were cloned into pBAD/HisB vector for recombinant protein expression in E. coli and protein purification using Ni-NTA chromatography with His-tag fused to AnCDA in N-terminal.
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This basic part is used for achieving bacterial cellulose-chitosan polymer by enzymatic reaction of deacetylation from bacterial cellulose-chitin. In addition, we have created a construct [https://parts.igem.org/Part:BBa_K4719019, BBa_K4719019] to improve the degree of deacetylation.
 
This basic part is used for achieving bacterial cellulose-chitosan polymer by enzymatic reaction of deacetylation from bacterial cellulose-chitin. In addition, we have created a construct [https://parts.igem.org/Part:BBa_K4719019, BBa_K4719019] to improve the degree of deacetylation.

Revision as of 20:16, 21 September 2023


AnCDA chitin deacetylase
Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 348
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 361
    Illegal XhoI site found at 593
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

Vilnius-Lithuania iGEM 2023 team's goal was to create a universal synthetic biology system for Komagataeibacter xylinus for in vivo bacterial cellulose polymer composition modification. Firstly, we chose to produce a cellulose-chitin polymer that would later be deacetylated, creating bacterial cellulose-chitosan. This polymer is an easily modifiable platform when compared to bacterial cellulose. The enhanced chemical reactivity of the bacterial cellulose-chitosan polymer allows for specific functionalizations in the biomedicine field, such as scaffold design. As a second approach, we designed indigo-dyed cellulose that could be used as a green chemistry way to apply cellulose in the textile industry. Lastly, we have achieved a composite of bacterial cellulose and polyhydroxybutyrate (PHB), which is synthesized by K. xylinus.

Usage and biology

AnCDA is chitin deacetylase from Aspergilus nidulans FGSCA4. This deacetylase is a member of carbohydrate esterase family 4 (CE4) of the CAZy database ([http://www.cazy.org/, www.cazy.org]). It has broad substrate specifity – AnCDA hydrolyzes N-acetamido groups in chitin oligomers, crystalline chitin and chitosan, however, it does not deacetylase peptidoglycan. AnCDA reaches its highest level of activity at approximately 50°C and pH 8.0 when Co2+ is used as a cofactor.

AnCDA gene has three exons and encodes the primary product of 237 amino acids, including the N-terminal extracellular signal sequence. Coding regions except leader sequence were cloned into pBAD/HisB vector for recombinant protein expression in E. coli and protein purification using Ni-NTA chromatography with His-tag fused to AnCDA in N-terminal.

This basic part is used for achieving bacterial cellulose-chitosan polymer by enzymatic reaction of deacetylation from bacterial cellulose-chitin. In addition, we have created a construct BBa_K4719019 to improve the degree of deacetylation.