Difference between revisions of "Part:BBa K4719002"

Revision as of 15:56, 19 September 2023

NAG5

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.

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.

@@ Line 13: / Line 13: @@
 ===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 [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].
+''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].
@@ Line 27: / Line 27: @@
 ===References===
-.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.
+.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.