Part:BBa_K4719002

NAG5

Introduction

Vilnius Lithuania iGEM 2023 team's goal was to create a universal synthetic biology system in 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 bacterial cellulose-chitosan polymer allows for specific functionalizations in the biomedicine field, such as scaffold design.

Bacterial cellulose-chitin polymer was achieved by increasing the production of UDP-N-acetylglucosamine, which can be recognized as a viable substrate for cellulose synthase and incorporated in the bacterial cellulose polymer. We employed two strategies to produce this material. The first approach was to add N-acetylglucosamine into the growth medium BBa_K4719013, and the second one was the production of N-acetylglucosamine by K. xylinus from simple sugars such as glucose, fructose, and saccharose in the growth medium BBa_K4719014.

Usage and Biology

NAG5 is N-acetylglucosamine kinase. The protein sequence is from Candida Albicans. This protein is a component of the N-acetylglucosamine catabolic cascade that phosphorylates N-acetylglucosamine (GlcNAc) and allows the unique ability to utilize GlcNAc as a carbon source. This part is used in BBa_K4719013. The function N-acetylglucosamine kinase has in our transcriptional unit is to convert extracellular N-acetylglucosamine into N-acetylglucosamine-6-phosphate, which is used as a substrate by N-acetylglucosamine kinase BBa_K4719001.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 198
  Illegal SpeI site found at 985
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 198
  Illegal SpeI site found at 985
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 198
  Illegal BamHI site found at 1489
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 198
  Illegal SpeI site found at 985
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 198
  Illegal SpeI site found at 985
  
• 1000
  COMPATIBLE WITH RFC[1000]