Part:BBa_K4719015
phaB
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. As the final approach, we have achieved a composite of bacterial cellulose and polyhydroxybutyrate (PHB), which is synthesized by K. xylinus.
We produced bacterial cellulose - PHB composite by introducing PHB synthesis operon into K. xylinus BBa_K4719017. The bacteria simultaneously produce both polymers, which are combined into the same material during the purification process.
Usage and Biology
PhaB is a gene encoding acetoacetyl-CoA reductase. The function of this protein is to catalyse the NADPH-dependent reduction of acetoacetyl-CoA. This step is intermediate in polyhydroxyalkanoates(PHA) pathways (1). PHAs are biodegradable thermoplastics, that naturally occur as storage compounds in bacteria when limited by the lack of nutrients other than carbon. This gene was cloned from polyhydroxybutyrate synthesis operon in the plasmid pBHR68 (2).
Characterisation
qPCR analysis of protein expression.
References
1.Kim J, Chang JH, Kim EJ, Kim KJ, (2014) ‘Crystal structure of (R)-3-hydroxybutyryl-CoA dehydrogenase PhaB from Ralstonia eutropha’, Biochem Biophys Res Commun, 443(3):783-8. doi: 10.1016/j.bbrc.2013.10.150. Epub 2013 Nov 6. PMID: 24211201.
2.Spiekermann, P. et al. (1999) ‘A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds’, Archives of Microbiology, 171(2), pp. 73–80. doi:10.1007/s002030050681.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 478
- 1000COMPATIBLE WITH RFC[1000]
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