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Characterization and Homology Modeling of Catalytically Active Recombinant PhaC_Ap Protein from Arthrospira platensis.

ABSTRACT: Polyhydroxybutyrate (PHB) is a biocompatible and biodegradable polymer that has the potential to replace fossil-derived polymers. The enzymes involved in the biosynthesis of PHB are β-ketothiolase (PhaA), acetoacetyl-CoA reductase (PhaB), and PHA synthase (PhaC). PhaC in Arthrospira platensis is the key enzyme for PHB production. In this study, the recombinant E. cloni^®10G cells harboring A. platensis phaC (rPhaC_Ap) was constructed. The overexpressed and purified rPhaC_Ap with a predicted molecular mass of 69 kDa exhibited V_max, K_m, and k_cat values of 24.5 ± 2 μmol/min/mg, 31.3 ± 2 µM and 412.7 ± 2 1/s, respectively. The catalytically active rPhaC_Ap was a homodimer. The three-dimensional structural model for the asymmetric PhaC_Ap homodimer was constructed based on Chromobacterium sp. USM2 PhaC (PhaC_Cs). The obtained model of PhaC_Ap revealed that the overall fold of one monomer was in the closed, catalytically inactive conformation whereas the other monomer was in the catalytically active, open conformation. In the active conformation, the catalytic triad residues (Cys151-Asp310-His339) were involved in the binding of substrate 3HB-CoA and the CAP domain of PhaC_Ap involved in the dimerization.

SUBMITTER: Duangsri C

PROVIDER: S-EPMC10215190 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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Characterization and Homology Modeling of Catalytically Active Recombinant PhaCAp Protein from Arthrospira platensis.

Duangsri Chanchanok C Salminen Tiina A TA Alix Marion M Kaewmongkol Sarawan S Akrimajirachoote Nattaphong N Khetkorn Wanthanee W Jittapalapong Sathaporn S Mäenpää Pirkko P Incharoensakdi Aran A Raksajit Wuttinun W

Biology 20230520 5

Polyhydroxybutyrate (PHB) is a biocompatible and biodegradable polymer that has the potential to replace fossil-derived polymers. The enzymes involved in the biosynthesis of PHB are β-ketothiolase (PhaA), acetoacetyl-CoA reductase (PhaB), and PHA synthase (PhaC). PhaC in Arthrospira platensis is the key enzyme for PHB production. In this study, the recombinant E. cloni®10G cells harboring A. platensis phaC (rPhaCAp) was constructed. The overexpressed an ...[more]

PMID: 37237563

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Project description:Background Arthrospira platensis is farmed worldwide due to its nutrient-rich properties and provides multiple benefits to human health. However, the wide usage of silver nanoparticles (Ag NPs) causes pollution which may affect the nutritional quality of A. platensis. Hence, this study aimed to investigate the interaction and accumulation of Ag NPs on A. platensis, and determine the changes in biomass and nutritional value of A. platensis due to the exposure to Ag NPs. Methods The interaction and accumulation of Ag NPs on A. platensis were examined through Fourier transformed infrared (FTIR) spectroscopy and scanning electron microscope (SEM). The loss in biomass together with the macromolecules, pigments, and phenolic compounds of A. platensis was investigated upon treating with various concentrations of Ag NPs (5, 10, 25, 50 and 100 µg/mL) for 24, 48, 72 and 96 h. Results The results showed that the treatment of A. platensis with Ag NPs caused a dose and time-dependent reduction in biomass, macronutrients, pigments and phenolic compounds. The highest detrimental effects were found at 96 h with the reported values of 65.71 ± 2.79%, 67.21 ± 3.98%, 48.99 ± 4.39% and 59.62 ± 3.96% reduction in biomass, proteins, carbohydrates and lipids, respectively, along with 82.99 ± 7.81%, 67.55 ± 2.63%, 75.03 ± 1.55%, and 63.43 ± 2.89% loss in chlorophyll-a, carotenoids, C-phycocyanin, and total phenolic compounds of A. platensis for 100 µg/mL of Ag NPs. The EDX analysis confirmed the surface accumulation of Ag NPs on Arthrospira cells, while SEM images evidenced the surface alterations and damage of the treated cells. The functional groups such as hydroxyl, amine, methyl, amide I, amide II, carboxyl, carbonyl and phosphate groups from the cell wall of the A. platensis were identified to be possibly involved in the interaction of Ag NPs with A. platensis. Conclusion The study confirmed that the exposure of Ag NPs is detrimental to A. platensis where the interaction and accumulation of Ag NPs on A. platensis caused reduction in biomass, macromolecules, pigments, and total phenolic compounds.

Dataset Information

Characterization and Homology Modeling of Catalytically Active Recombinant PhaC_Ap Protein from Arthrospira platensis.

Publications

Characterization and Homology Modeling of Catalytically Active Recombinant PhaC<sub>Ap</sub> Protein from <i>Arthrospira platensis</i>.

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