Difference between revisions of "Part:BBa K4719004"

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===Introduction===
 
===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.  
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<b>Vilnius-Lithuania iGEM 2023</b> team's goal was to create <b> synthetic biology tools for <i>in vivo</i> alterations of <i>Komagataeibacter xylinus</i> bacterial cellulose polymer composition</b>. Firstly, we chose to produce a <b>cellulose-chitin copolymer</b> that would later be deacetylated, creating <b>bacterial cellulose-chitosan</b>. 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 <b>indigo-dyed cellulose</b> that could be used as a green chemistry way to apply cellulose in the textile industry. Lastly, we have achieved a of <b>bacterial cellulose and polyhydroxybutyrate (PHB) composite</b>, which is synthesized by <i>K. xylinus</i>.
 
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Revision as of 20:25, 9 October 2023


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


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]


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.