Difference between revisions of "Part:BBa K4719002"
 
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===Introduction===
 
===Introduction===
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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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===Usage and Biology===
 
===Usage and Biology===
  
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''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 [https://parts.igem.org/Part:BBa_K4719001#references (1)]. This part is used in [https://parts.igem.org/Part:BBa_K4719013 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 [https://parts.igem.org/Part:BBa_K4719001 BBa_K4719001].
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''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 [https://parts.igem.org/Part:BBa_K4719002#references (1)]. This part is used in [https://parts.igem.org/Part:BBa_K4719013 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 [https://parts.igem.org/Part:BBa_K4719001 BBa_K4719001].
  
  
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===References===
 
===References===
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1.Mio, T. et al. (2000) ‘Functional cloning and mutational analysis of the human cDNA for phosphoacetylglucosamine mutase: identification of the amino acid residues essential for the catalysis’, Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1492(2–3), pp. 369–376. doi:10.1016/s0167-4781(00)00120-2.
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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.

Latest revision as of 20:21, 9 October 2023


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.