Difference between revisions of "Part:BBa K4719002"

Revision as of 15:52, 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

References

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.