Abstract
Objectives
To engineer a recombinant Escherichia coli to produce polyhydroxyalkanoate biopolymer beads displaying carbonic anhydrase (CA) from Desulfovibrio vulgaris str. “Miyazaki F” (DvCA).
Results
The highest measured specific activity of the immobilised CA was 211 U/mg DvCA. The immobilised CA was thermostable, retaining 114 U/mg DvCA of activity after incubation at 90 °C for 1 h. Additionally, the immobilised CA tolerated 30 min incubations in a variety of pH conditions and was especially tolerant of alkaline conditions, retaining 131 U/mg DvCA of activity after pH 12 incubation.
Conclusion
CA has a potential role in atmospheric CO2 mitigation strategies and the stability of the functionalised beads indicates suitability for use in industrial settings such as coal-fired power plants.
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Acknowledgments
This study was funded by Institute of Fundamental Sciences (Massey University) and the MacDiarmid Institute. DOH received funding from the Institute of Fundamental Sciences at Massey University and a Massey University PhD scholarship.
Supplementary information
Supplementary Table 1—Point mutations in the enhanced Desulfovibrio vulgaris str. “Miyazaki F” carbonic anhydrase.
Supplementary Table 2—Tryptic peptides of PhaC fusion proteins as identified by MALDI-TOF/TOF MS. Total bead protein was electrophoresed onto a gel and a band corresponding to the theoretical molecular weight of the fusion protein was isolated and analysed by mass spectrometry.
Supplementary Fig. 1—PHB content of Escherichia coli BL21 blue cells as measured by GC/MS. Cells contained either PhaC or PhaC-fusion expression plasmids and were cultivated for 48 h at 25 °C after induction with isopropyl-β-D-1-thiogalactopyranoside.
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Hooks, D.O., Rehm, B.H.A. Surface display of highly-stable Desulfovibrio vulgaris carbonic anhydrase on polyester beads for CO2 capture. Biotechnol Lett 37, 1415–1420 (2015). https://doi.org/10.1007/s10529-015-1803-7
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DOI: https://doi.org/10.1007/s10529-015-1803-7