期刊
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷 186, 期 -, 页码 414-423出版社
ELSEVIER
DOI: 10.1016/j.ijbiomac.2021.07.041
关键词
Polyhydroxyalkanoate (PHA); Aquitalea sp.; PHA synthase (PhaC) truncation; Substrate specificity; PHA granule morphology; Dimerization
资金
- Ministry of Higher Education Malaysia [203/PBIOLOGI/67811001]
- JST ASTEP [29A1027]
- NEDO [P20005]
- Science and Technology Research Partnership for Sustainable Development (SATREPS)
The study demonstrated the importance of both the N- and C-termini of PhaC1(As) in regulating intracellular granule morphology, activity, substrate specificity, dimerization, and stability of the synthase.
Polyhydroxyalkanoates (PHAs) are biopolyesters synthesized by microorganisms as intracellular energy reservoirs under stressful environmental conditions. PHA synthase (PhaC) is the key enzyme responsible for PHA biosynthesis, but the importance of its N- and C-terminal ends still remains elusive. Six plasmid constructs expressing truncation variants of Aquitalea sp. USM4 PhaC (PhaC1(As)) were generated and heterologously expressed in Cupriavidus necator PHB(-)4. Removal of the first six residues at the N-terminus enabled the modulation of PHA composition without altering the PHA content in cells. Meanwhile, deletion of 13 amino acids from the C-terminus greatly affected the catalytic activity of PhaC1(As), retaining only 1.1-7.4% of the total activity. Truncation(s) at the N- and/or C-terminus of PhaC1(As) gradually diminished the incorporation of comonomer units, and revealed that the N-terminal region is essential for PhaC1(As) dimerization whereas the C-terminal region is required for stabilization. Notably, transmission electron microscopy analysis showed that PhaC modification affected the morphology of intracellular PHA granules, which until now is only known to be regulated by phasins. This study provided substantial evidence and highlighted the significance of both the N-and C-termini of PhaC1(As) in regulating intracellular granule morphology, activity, substrate specificity, dimerization and stability of the synthase.
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