Coding

Part:BBa_K4719002

Designed by: Auguste Stankeviciute   Group: iGEM23_Vilnius-Lithuania   (2023-08-30)
Revision as of 20:21, 9 October 2023 by Augustestankeviciute (Talk | contribs)

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NAG5

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 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 (1). 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]


References

1.Wendland, J., Schaub, Y. and Walther, A. (2009) ‘N-Acetylglucosamine Utilization by Saccharomyces cerevisiae Based on Expression of Candida albicans NAG Genes’, Applied and Environmental Microbiology, 75(18), pp. 5840–5845. doi:10.1128/aem.00053-09.

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