Coding

Part:BBa_K4719005

Designed by: Auguste Stankeviciute   Group: iGEM23_Vilnius-Lithuania   (2023-08-30)


GNA1

Introduction

Vilnius-Lithuania iGEM 2023 team's goal was to create synthetic biology tools for in vivo alterations of Komagataeibacter xylinus bacterial cellulose polymer composition. Firstly, we chose to produce a cellulose-chitin copolymer 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 of bacterial cellulose and polyhydroxybutyrate (PHB) composite, which is synthesized by K. xylinus.

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 other sugars such as glucose, fructose, and saccharose in the growth medium BBa_K4719014.

Usage and Biology

GNA1 is glucosamine 6-phosphate N-acetyltransferase. This enzyme catalyzes the transfer of an acetyl group from acetyl coenzyme A to glucosamine-6-phosphate to form N-acetylglucosamine-6-phosphate, which is an essential intermediate in UDP-GlcNAc biosynthesis(1) . GNA1 is a part in BBa_K4719014.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 118
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 118
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 118
    Illegal BamHI site found at 457
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 118
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 118
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1.Mio, T. et al. (1999) ‘Saccharomyces cerevisiae GNA1, an Essential Gene Encoding a Novel Acetyltransferase Involved in UDP-N-acetylglucosamine Synthesis’, Journal of Biological Chemistry, 274(1), pp. 424–429. doi:10.1074/jbc.274.1.424.

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