Part:BBa_K4719004
GFA1
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 (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 other sugars such as glucose, fructose, and saccharose in the growth medium BBa_K4719014.
Usage and Biology
GFA1 is glutamine-fructose-6-phosphate aminotransferase. The protein sequence is from Candida albicans. This enzyme catalyzes the formation of glucosamine-6-phosphate and glutamate from fructose-6-phosphate and glutamine (1). 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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 361
Illegal EcoRI site found at 859
Illegal XbaI site found at 1459 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 361
Illegal EcoRI site found at 859 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 361
Illegal EcoRI site found at 859
Illegal BglII site found at 1318 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 361
Illegal EcoRI site found at 859
Illegal XbaI site found at 1459 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 361
Illegal EcoRI site found at 859
Illegal XbaI site found at 1459 - 1000COMPATIBLE WITH RFC[1000]
References
1.Dummitt, B., Micka, W.S. and Chang, Y.-H. (2005) ‘Yeast Glutamine-fructose-6-phosphate Aminotransferase (Gfa1) Requires Methionine Aminopeptidase Activity for Proper Function’, Journal of Biological Chemistry, 280(14), pp. 14356–14360. doi:10.1074/jbc.m501059200.
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