Part:BBa_K4719002
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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 198
Illegal SpeI site found at 985 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 198
Illegal SpeI site found at 985 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 198
Illegal BamHI site found at 1489 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 198
Illegal SpeI site found at 985 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 198
Illegal SpeI site found at 985 - 1000COMPATIBLE 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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