Part:BBa_K4719004

GFA1

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 (UDP-GlcNAc), which can be recognized as a viable substrate for cellulose synthase and incorporated in 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

GFA1 is glutamine-fructose-6-phosphate aminotransferase. This enzyme catalyzes the formation of glucosamine-6-phosphate and glutamate from fructose-6-phosphate and glutamine. This is a crucial step in our system, that produces bacterial cellulose-chitin polymer from simple sugars in the growth medium. GFA1 is used as a part in BBa_K4719014.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 361
  Illegal EcoRI site found at 859
  Illegal XbaI site found at 1459
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 361
  Illegal EcoRI site found at 859
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 361
  Illegal EcoRI site found at 859
  Illegal BglII site found at 1318
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 361
  Illegal EcoRI site found at 859
  Illegal XbaI site found at 1459
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 361
  Illegal EcoRI site found at 859
  Illegal XbaI site found at 1459
  
• 1000
  COMPATIBLE WITH RFC[1000]